Immunoglobulin protease variants and uses thereof

Abstract

Embodiments provided herein, provide for polypeptides and molecules comprising a polypeptide having protease activity, which can be linked to a Fc molecule, or half-life extension moiety, pharmaceutical compositions comprising the same, and methods that can be used to treat disorders, such as immunoglobulin mediated disorders, including autoimmune disorders and diseases.

Description

[0001] DOCKET NO. SES-020WO PATENT

[0002] IMMUNOGLOBULIN PROTEASE VARIANTS AND USES THEREOF

[0003] CROSS-REFERENCE TO RELATED APPLICATIONS

[0004] This application claims priority to U.S. Provisional Application No. 63 / 716,198, filed November 4, 2024, which is hereby incorporated by reference in its entirety.

[0005] REFERENCE TO SEQUENCE LISTING SUBMITTED ELECTRONICALLY

[0006] The instant application contains a Sequence Listing which has been submitted electronically in XML file format and is hereby incorporated by reference in its entirety. Said XML copy, created on November 4, 2025, is named “SES-020WO_SEQ.xml” and is 255,416 bytes in size.

[0007] FIELD

[0008] The embodiments provided herein relate to polypeptides comprising a polypeptide having protease activity and, optionally, an Fc moiety, or a half-life extension moiety, and compositions comprising the same.

[0009] BACKGROUND

[0010] Immunoglobulin G-degrading enzyme of S. pyogenes (IdeS) is an extracellular cysteine protease produced by the human pathogen S. pyogenes. IdeS is a virulence factor of S. pyogenes, which is responsible for common infections like tonsillitis and strep throat. IdeS has an extraordinarily high degree of substrate specificity, with its only identified substrate being immunoglobulin G (IgG). IdeS catalyzes a single proteolytic cleavage in the lower hinge region of the heavy chains of all subclasses of human IgG. IdeS also catalyzes an equivalent cleavage of the heavy chains of some subclasses of IgG in various animals. IdeS efficiently cleaves IgG to Fc and F(ab')2 fragments. Certain IgG cysteine proteases have been identified, but can be immunogenic when administered to humans or other animals. The immunogenicity of the proteases can limit the ability to administer the proteases multiple times. However, pathogenic IgG antibodies constitute an important clinical problem contributing to the pathogenesis of a number of autoimmune conditions and acute transplant rejection. To be able to effectively reduce, or eliminate such antibodies is therefore an important clinical challenge. The embodiments provided for herein fulfill this need as well as others.

[0011] -1-

[0012] IPTS / 200163498.2 DOCKET NO. SES-020WO PATENT

[0013] SUMMARY

[0014] Provided herein is a polypeptide comprising an amino acid sequence having at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity to SEQ ID NO: 35, and having a set of mutations selected from: a threonine (T) at position 60, an aspartic acid (D) at position 61, an isoleucine (I) at position 63, a tyrosine (Y) at position 64, a leucine (L) at position 69, a lysine (K) at position 74, a glycine (G) at position 76, a leucine (L) at position 85, a glutamic acid (E) at position 112, an asparagine (N) at position 116, a lysine (K) at position 119, a lysine (K) at position 120, a tyrosine (Y) at position 121, a glycine (G) at position 130, a leucine (L) at position 134, an aspartic acid (D) at position 145, a serine (S) at position 146, a threonine (T) at position 148, an aspartic acid (D) at position 154, an aspartic acid (D) at position 190, an arginine (R) at position 198, an asparagine (N) at position 223, a tyrosine (Y) at position 225, a leucine (L) at position 227, a threonine (T) at position 231, a lysine (K) at position 241, an isoleucine (I) at position 251, a glycine (G) at position 261, an alanine (A) at position 273, a glutamic acid (E) at position 274, a glutamic acid (E) at position 278, a proline (P) at position 289, a leucine (L) at position 303, a glutamic acid (E) at position 314, and an isoleucine (I) at position 334, wherein the positions correspond to the numbering as set forth in SEQ ID NO: 35; a threonine (T) at position 60, an aspartic acid (D) at position 61, an isoleucine (I) at position 63, a tyrosine (Y) at position 64, a leucine (L) at position 69, a lysine (K) at position 74, a glycine (G) at position 76, a leucine (L) at position 85, a glutamic acid (E) at position 112, an asparagine (N) at position 116, a lysine (K) at position 119, a lysine (K) at position 120, a tyrosine (Y) at position 121, a glycine (G) at position 130, a leucine (L) at position 134, an aspartic acid (D) at position 145, a serine (S) at position 146, a threonine (T) at position 148, an aspartic acid (D) at position 154, an aspartic acid (D) at position 163, an aspartic acid (D) at position 190, an arginine (R) at position 198, an asparagine (N) at position 223, a tyrosine (Y) at position 225, a leucine (L) at position 227, a threonine (T) at position 231, a lysine (K) at position 241, an isoleucine (I) at position 251, a glycine (G) at position 261, an alanine (A) at position 273, a glutamic acid (E) at position 274, a glutamic acid (E) at position 278, a proline (P) at position 289, a leucine (L) at position 303, a glutamic acid (E) at position 314, and an isoleucine (I) at position 334, wherein the positions correspond to the numbering as set forth in SEQ ID NO: 35;

[0015] -2-

[0016] IPTS / 200163498.2 DOCKET NO. SES-020WO PATENT a threonine (T) at position 60, an aspartic acid (D) at position 61, an isoleucine (I) at position 63, a tyrosine (Y) at position 64, a leucine (L) at position 69, a lysine (K) at position 74, a glycine (G) at position 76, a leucine (L) at position 85, a glutamic acid (E) at position 112, an asparagine (N) at position 116, a lysine (K) at position 119, a lysine (K) at position 120, a tyrosine (Y) at position 121, a glycine (G) at position 130, a leucine (L) at position 134, an aspartic acid (D) at position 145, a serine (S) at position 146, a threonine (T) at position 148, an aspartic acid (D) at position 154, an asparagine (N) at position 163, an alanine (A) at position 166, an arginine (R) at position 167, a glutamine (Q) at position 168, an aspartic acid (D) at position 190, an arginine (R) at position 198, an asparagine (N) at position 223, a tyrosine (Y) at position 225, a leucine (L) at position 227, a threonine (T) at position 231, a lysine (K) at position 241, an isoleucine (I) at position 251, a glycine (G) at position 261, an alanine (A) at position 273, a glutamic acid (E) at position 274, a glutamic acid (E) at position 278, a proline (P) at position 289, a leucine (L) at position 303, a glutamic acid (E) at position 314, and an isoleucine (I) at position 334, wherein the positions correspond to the numbering as set forth in SEQ ID NO: 35; or a threonine (T) at position 60, an aspartic acid (D) at position 61, an isoleucine (I) at position 63, a tyrosine (Y) at position 64, a leucine (L) at position 69, a lysine (K) at position 74, a glycine (G) at position 76, a leucine (L) at position 85, a glutamic acid (E) at position 112, an asparagine (N) at position 116, a lysine (K) at position 119, a lysine (K) at position 120, a tyrosine (Y) at position 121, a glycine (G) at position 130, a leucine (L) at position 134, an aspartic acid (D) at position 145, a serine (S) at position 146, a threonine (T) at position 148, an aspartic acid (D) at position 154, an asparagine (N) at position 163, an alanine (A) at position 166, an arginine (R) at position 167, a glutamine (Q) at position 168, an isoleucine (I) at position 169, an aspartic acid (D) at position 190, an arginine (R) at position 198, an asparagine (N) at position 223, a tyrosine (Y) at position 225, a leucine (L) at position 227, a threonine (T) at position 231, a lysine (K) at position 241, an isoleucine (I) at position 251, a glycine (G) at position 261, an alanine (A) at position 273, a glutamic acid (E) at position 274, a glutamic acid (E) at position 278, a proline (P) at position 289, a leucine (L) at position 303, a glutamic acid (E) at position 314, and an isoleucine (I) at position 334, wherein the positions correspond to the numbering as set forth in SEQ ID NO: 35.

[0017] In some embodiments, provided herein is a polypeptide comprising: an amino acid sequence having at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at

[0018] -3-

[0019] IPTS / 200163498.2 DOCKET NO. SES-020WO PATENT least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity to SEQ ID NO: 57, provided that the polypeptide comprises a threonine (T) at position 60, an aspartic acid (D) at position 61, an isoleucine (I) at position 63, a tyrosine (Y) at position 64, a leucine (L) at position 69, a lysine (K) at position 74, a glycine (G) at position 76, a leucine (L) at position 85, a glutamic acid (E) at position 112, an asparagine (N) at position 116, a lysine (K) at position 119, a lysine (K) at position 120, a tyrosine (Y) at position 121, a glycine (G) at position 130, a leucine (L) at position 134, an aspartic acid (D) at position 145, a serine (S) at position 146, a threonine (T) at position 148, an aspartic acid (D) at position 154, an aspartic acid (D) at position 190, an arginine (R) at position 198, an asparagine (N) at position 223, a tyrosine (Y) at position 225, a leucine (L) at position 227, a threonine (T) at position 231, a lysine (K) at position 241, an isoleucine (I) at position 251, a glycine (G) at position 261, an alanine (A) at position 273, a glutamic acid (E) at position 274, a glutamic acid (E) at position 278, a proline (P) at position 289, a leucine (L) at position 303, a glutamic acid (E) at position 314, and an isoleucine (I) at position 334, wherein the positions correspond to the numbering as set forth in SEQ ID NO: 35; an amino acid sequence having at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity to SEQ ID NO: 58, provided that the polypeptide comprises a threonine (T) at position 60, an aspartic acid (D) at position 61, an isoleucine (I) at position 63, a tyrosine (Y) at position 64, a leucine (L) at position 69, a lysine (K) at position 74, a glycine (G) at position 76, a leucine (L) at position 85, a glutamic acid (E) at position 112, an asparagine (N) at position 116, a lysine (K) at position 119, a lysine (K) at position 120, a tyrosine (Y) at position 121, a glycine (G) at position 130, a leucine (L) at position 134, an aspartic acid (D) at position 145, a serine (S) at position 146, a threonine (T) at position 148, an aspartic acid (D) at position 154, an aspartic acid (D) at position 163, an aspartic acid (D) at position 190, an arginine (R) at position 198, an asparagine (N) at position 223, a tyrosine (Y) at position 225, a leucine (L) at position 227, a threonine (T) at position 231, a lysine (K) at position 241, an isoleucine (I) at position 251, a glycine (G) at position 261, an alanine (A) at position 273, a glutamic acid (E) at position 274, a glutamic acid (E) at position 278, a proline (P) at position 289, a leucine (L) at position 303, a glutamic acid (E) at position 314, and an isoleucine (I) at position 334, wherein the positions correspond to the numbering as set forth in SEQ ID NO: 35; an amino acid sequence having at

[0020] -4-

[0021] IPTS / 200163498.2 DOCKET NO. SES-020WO PATENT least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity to SEQ ID NO: 59, provided that the polypeptide comprises a threonine (T) at position 60, an aspartic acid (D) at position 61, an isoleucine (I) at position 63, a tyrosine (Y) at position 64, a leucine (L) at position 69, a lysine (K) at position 74, a glycine (G) at position 76, a leucine (L) at position 85, a glutamic acid (E) at position 112, an asparagine (N) at position 116, a lysine (K) at position 119, a lysine (K) at position 120, a tyrosine (Y) at position 121, a glycine (G) at position 130, a leucine (L) at position 134, an aspartic acid (D) at position 145, a serine (S) at position 146, a threonine (T) at position 148, an aspartic acid (D) at position 154, an asparagine (N) at position 163, an alanine (A) at position 166, an arginine (R) at position 167, a glutamine (Q) at position 168, an aspartic acid (D) at position 190, an arginine (R) at position 198, an asparagine (N) at position 223, a tyrosine (Y) at position 225, a leucine (L) at position 227, a threonine (T) at position 231, a lysine (K) at position 241, an isoleucine (I) at position 251, a glycine (G) at position 261, an alanine (A) at position 273, a glutamic acid (E) at position 274, a glutamic acid (E) at position 278, a proline (P) at position 289, a leucine (L) at position 303, a glutamic acid (E) at position 314, and an isoleucine (I) at position 334, wherein the positions correspond to the numbering as set forth in SEQ ID NO: 35; or an amino acid sequence having at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity to SEQ ID NO: 60, provided that the polypeptide comprises a threonine (T) at position 60, an aspartic acid (D) at position 61, an isoleucine (I) at position 63, a tyrosine (Y) at position 64, a leucine (L) at position 69, a lysine (K) at position 74, a glycine (G) at position 76, a leucine (L) at position 85, a glutamic acid (E) at position 112, an asparagine (N) at position 116, a lysine (K) at position 119, a lysine (K) at position 120, a tyrosine (Y) at position 121, a glycine (G) at position 130, a leucine (L) at position 134, an aspartic acid (D) at position 145, a serine (S) at position 146, a threonine (T) at position 148, an aspartic acid (D) at position 154, an asparagine (N) at position 163, an alanine (A) at position 166, an arginine (R) at position 167, a glutamine (Q) at position 168, an isoleucine (I) at position 169, an aspartic acid (D) at position 190, an arginine (R) at position 198, an asparagine (N) at position 223, a tyrosine (Y) at position 225, a leucine (L) at position 227, a threonine (T) at position 231, a lysine (K) at

[0022] -5-

[0023] IPTS / 200163498.2 DOCKET NO. SES-020WO PATENT position 241, an isoleucine (I) at position 251, a glycine (G) at position 261, an alanine (A) at position 273, a glutamic acid (E) at position 274, a glutamic acid (E) at position 278, a proline (P) at position 289, a leucine (L) at position 303, a glutamic acid (E) at position 314, and an isoleucine (I) at position 334, wherein the positions correspond to the numbering as set forth in SEQ ID NO: 35.

[0024] In some embodiments, provided herein is a polypeptide comprising an amino acid of any one of SEQ ID NOs: 57, 58, 59, or 60.

[0025] In some embodiments, provided herein is a polypeptide comprising: an amino acid sequence having at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity to SEQ ID NO: 57, provided that the polypeptide comprises a threonine (T) at position 60, an aspartic acid (D) at position 61, an isoleucine (I) at position 63, a tyrosine (Y) at position 64, a leucine (L) at position 69, a lysine (K) at position 74, a glycine (G) at position 76, a leucine (L) at position 85, a glutamic acid (E) at position 112, an asparagine (N) at position 116, a lysine (K) at position 119, a lysine (K) at position 120, a tyrosine (Y) at position 121, a glycine (G) at position 130, a leucine (L) at position 134, an aspartic acid (D) at position 145, a serine (S) at position 146, a threonine (T) at position 148, an aspartic acid (D) at position 154, an aspartic acid (D) at position 190, an arginine (R) at position 198, an asparagine (N) at position 223, a tyrosine (Y) at position 225, a leucine (L) at position 227, a threonine (T) at position 231, a lysine (K) at position 241, an isoleucine (I) at position 251, a glycine (G) at position 261, an alanine (A) at position 273, a glutamic acid (E) at position 274, a glutamic acid (E) at position 278, a proline (P) at position 289, a leucine (L) at position 303, a glutamic acid (E) at position 314, and an isoleucine (I) at position 334, wherein the positions correspond to the numbering as set forth in SEQ ID NO: 35; and an Fc polypeptide, wherein the polypeptide is conjugated to the Fc polypeptide via a linker, such as a peptide linker.

[0026] In some embodiments, provided herein is a polypeptide comprising: an amino acid sequence having at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity to SEQ ID NO: 58,

[0027] -6-

[0028] IPTS / 200163498.2 DOCKET NO. SES-020WO PATENT provided that the polypeptide comprises a threonine (T) at position 60, an aspartic acid (D) at position 61, an isoleucine (I) at position 63, a tyrosine (Y) at position 64, a leucine (L) at position 69, a lysine (K) at position 74, a glycine (G) at position 76, a leucine (L) at position 85, a glutamic acid (E) at position 112, an asparagine (N) at position 116, a lysine (K) at position 119, a lysine (K) at position 120, a tyrosine (Y) at position 121, a glycine (G) at position 130, a leucine (L) at position 134, an aspartic acid (D) at position 145, a serine (S) at position 146, a threonine (T) at position 148, an aspartic acid (D) at position 154, an aspartic acid (D) at position 163, an aspartic acid (D) at position 190, an arginine (R) at position 198, an asparagine (N) at position 223, a tyrosine (Y) at position 225, a leucine (L) at position 227, a threonine (T) at position 231, a lysine (K) at position 241, an isoleucine (I) at position 251, a glycine (G) at position 261, an alanine (A) at position 273, a glutamic acid (E) at position 274, a glutamic acid (E) at position 278, a proline (P) at position 289, a leucine (L) at position 303, a glutamic acid (E) at position 314, and an isoleucine (I) at position 334, wherein the positions correspond to the numbering as set forth in SEQ ID NO: 35; and an Fc polypeptide, wherein the polypeptide is conjugated to the Fc polypeptide via a linker, such as a peptide linker.

[0029] In some embodiments, provided herein is a polypeptide comprising: an amino acid sequence having at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity to SEQ ID NO: 59, provided that the polypeptide comprises a threonine (T) at position 60, an aspartic acid (D) at position 61, an isoleucine (I) at position 63, a tyrosine (Y) at position 64, a leucine (L) at position 69, a lysine (K) at position 74, a glycine (G) at position 76, a leucine (L) at position 85, a glutamic acid (E) at position 112, an asparagine (N) at position 116, a lysine (K) at position 119, a lysine (K) at position 120, a tyrosine (Y) at position 121, a glycine (G) at position 130, a leucine (L) at position 134, an aspartic acid (D) at position 145, a serine (S) at position 146, a threonine (T) at position 148, an aspartic acid (D) at position 154, an asparagine (N) at position 163, an alanine (A) at position 166, an arginine (R) at position 167, a glutamine (Q) at position 168, an aspartic acid (D) at position 190, an arginine (R) at position 198, an asparagine (N) at position 223, a tyrosine (Y) at position 225, a leucine (L) at position 227, a threonine (T) at position 231, a lysine (K) at position 241, an isoleucine (I) at position 251, a glycine (G) at position 261, an alanine (A) at position 273, a glutamic acid (E) at position 274, a glutamic acid

[0030] -7-

[0031] IPTS / 200163498.2 DOCKET NO. SES-020WO PATENT

[0032] (E) at position 278, a proline (P) at position 289, a leucine (L) at position 303, a glutamic acid (E) at position 314, and an isoleucine (I) at position 334, wherein the positions correspond to the numbering as set forth in SEQ ID NO: 35; and an Fc polypeptide, wherein the polypeptide is conjugated to the Fc polypeptide via a linker, such as a peptide linker.

[0033] In some embodiments, provided herein is a polypeptide comprising: an amino acid sequence having at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity to SEQ ID NO: 60, provided that the polypeptide comprises a threonine (T) at position 60, an aspartic acid (D) at position 61, an isoleucine (I) at position 63, a tyrosine (Y) at position 64, a leucine (L) at position 69, a lysine (K) at position 74, a glycine (G) at position 76, a leucine (L) at position 85, a glutamic acid (E) at position 112, an asparagine (N) at position 116, a lysine (K) at position 119, a lysine (K) at position 120, a tyrosine (Y) at position 121, a glycine (G) at position 130, a leucine (L) at position 134, an aspartic acid (D) at position 145, a serine (S) at position 146, a threonine (T) at position 148, an aspartic acid (D) at position 154, an asparagine (N) at position 163, an alanine (A) at position 166, an arginine (R) at position 167, a glutamine (Q) at position 168, an isoleucine (I) at position 169, an aspartic acid (D) at position 190, an arginine (R) at position 198, an asparagine (N) at position 223, a tyrosine (Y) at position 225, a leucine (L) at position 227, a threonine (T) at position 231, a lysine (K) at position 241, an isoleucine (I) at position 251, a glycine (G) at position 261, an alanine (A) at position 273, a glutamic acid (E) at position 274, a glutamic acid (E) at position 278, a proline (P) at position 289, a leucine (L) at position 303, a glutamic acid (E) at position 314, and an isoleucine (I) at position 334, wherein the positions correspond to the numbering as set forth in SEQ ID NO: 35; and an Fc polypeptide, wherein the polypeptide is conjugated to the Fc polypeptide via a linker, such as a peptide linker.

[0034] In some embodiments, provided herein is a molecule comprising: a polypeptide comprising an amino acid sequence of any one of SEQ ID NOs: 57, 58, 59, or 60; and an Fc polypeptide, wherein the polypeptide is conjugated to the Fc polypeptide via a linker, such as a peptide linker.

[0035] In some embodiments, provided herein is a molecule comprising: an Fc polypeptide having an amino acid sequence of SEQ ID NO: 23; and a polypeptide having an amino acid

[0036] -8-

[0037] IPTS / 200163498.2 DOCKET NO. SES-020WO PATENT sequence of any one of SEQ ID NOs: 57, 58, 59, or 60, wherein the polypeptide is conjugated to the Fc polypeptide via a linker, such as a peptide linker.

[0038] In some embodiments, provided herein is a method of treating a subject having, or at risk, or elevated risk, for having, an IgG mediated disease or disorder, by administering a polypeptide having protease activity, the polypeptide comprising: a) means for proteolytically cleaving the IgG; and b) an Fc domain comprising: an amino acid sequence of SEQ ID NO: 19; an amino acid sequence of SEQ ID NO: 21; an amino acid sequence of SEQ ID NO: 22; an amino acid sequence of SEQ ID NO: 23; an amino acid sequence of SEQ ID NO: 24; or an amino acid sequence of SEQ ID NO: 25; wherein said polypeptide having protease activity has increased half-life.

[0039] BRIEF DESCRIPTION OF FIGURES

[0040] FIG. 1 illustrates a non-limiting example of a bivalent and a monovalent protease as provided for herein.

[0041] FIG. 2A shows IgGl cleavage products.

[0042] FIG. 2B shows IgGl cleavage product distributions.

[0043] FIG. 2C shows IgGl cleavage activity.

[0044] FIG. 3 A shows AC-SINS screening results for l-step purified protease variants.

[0045] FIG. 3B shows AC-SINS screening results for 2-step purified protease variants.

[0046] FIG. 4 shows polyreactivity ELISA results.

[0047] FIG. 5A shows binding of pre-existing ADA in IVIG from individual donor.

[0048] FIG. 5B shows binding of pre-existing ADA in plasma from individual donors.

[0049] FIG. 5C shows total IgG plasma cleavage.

[0050] DETAILED DESCRIPTION

[0051] This application incorporates by reference U.S. Provisional Application Nos. 63 / 384, 253, filed November 18, 2022, 63 / 478,789, filed January 6, 2023, 63 / 483,127, filed February 3, 2023, 63 / 483,142, filed February 3, 2023, 63 / 493,385, filed March 31, 2023, 63 / 493,450, filed

[0052] -9-

[0053] IPTS / 200163498.2 DOCKET NO. SES-020WO PATENT

[0054] March 31, 2023, 63 / 506,539, filed June 6, 2023, 63 / 600,157, filed November 17, 2023, 63 / 669,541, filed July 10, 2024, 63 / 701,666, filed October 1, 2024, U.S. Non-Provisional Application Nos. 18,512,556, filed November 17, 2023, now U.S. Patent No. 12,091,694, 18 / 405,861, filed January 5, 2024, now U.S. Patent No. 12,129,499, 18,815,585, filed August 26, 2024, 18 / 885,268, filed September 13, 2024, and International Application No.

[0055] PCT / US2023 / 080313, filed November 17, 2023, and PCT / US2024 / 010503, filed January 5, 2024, each of which is hereby incorporated by reference in its entirety

[0056] As used herein and in the appended claims, the singular forms “a”, “an” and “the” include plural reference unless the context clearly dictates otherwise.

[0057] As used herein, the term “about” means that the numerical value is approximate and small variations would not significantly affect the practice of the disclosed embodiments. Where a numerical limitation is used, unless indicated otherwise by the context, “about” means the numerical value can vary by ±5% and remain within the scope of the disclosed embodiments. Thus, about 100 means 95 to 105.

[0058] As used herein, the term “animal” includes, but is not limited to, humans and non-human vertebrates such as wild, domestic, and farm animals. As used herein, the term “mammal” means a rodent (i.e., a mouse, a rat, or a guinea pig), a monkey, a cat, a dog, a cow, a horse, a pig, or a human. In some embodiments, the mammal is a human.

[0059] As used herein, the term “contacting” means bringing together of two elements in an in vitro system or an in vivo system. For example, “contacting” a therapeutic compound with an individual or patient, which can be an animal or mammal, or cell includes the administration of the compound to an individual or patient, such as a human, as well as, for example, introducing a compound into a sample containing a cellular or purified preparation containing target.

[0060] As used herein, the terms “comprising” (and any form of comprising, such as “comprise”, “comprises”, and “comprised”), “having” (and any form of having, such as “have” and “has”), “including” (and any form of including, such as “includes” and “include”), or “containing” (and any form of containing, such as “contains” and “contain”), are inclusive or open-ended and do not exclude additional, unrecited elements, components, or method steps. Any composition or method that recites the term “comprising” should also be understood to also describe such

[0061] -10-

[0062] IPTS / 200163498.2 DOCKET NO. SES-020WO PATENT compositions as consisting, consisting of, or consisting essentially of the recited components or elements.

[0063] As used herein, the term “fused” or “linked” when used in reference to a protein or molecule having different domains or heterologous sequences means that the protein domains are part of the same peptide chain that are connected to one another with either peptide bonds or other covalent bonding. The domains or section can be linked or fused directly to one another or another domain or peptide sequence can be between the two domains or sequences and such sequences would still be considered to be fused or linked to one another.

[0064] As used herein, the term “individual,” “subject,” or “patient,” used interchangeably, means any animal, including mammals, such as mice, rats, other rodents, rabbits, dogs, cats, swine, cattle, sheep, horses, or primates, such as humans or as otherwise provided for herein.

[0065] As used herein, the term “inhibit” refers to a result, symptom, or activity being reduced as compared to the activity or result in the absence of the compound that is inhibiting the result, symptom, or activity. In some embodiments, the result, symptom, or activity, is inhibited by about, or, at least, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, or 99%. An result, symptom, or activity can also be inhibited if it is completely elimination or extinguished.

[0066] As used herein, the phrase “in need thereof’ means that the subject has been identified as having a need for the particular method or treatment and the method is performed with a specific intent of providing the treatment or performing the particular method. In some embodiments, the identification can be by any means of diagnosis. In any of the methods and treatments described herein, the subject can be in need thereof. In some embodiments, the subject is in an environment or will be traveling to an environment in which a particular disease, disorder, or condition is prevalent.

[0067] As used herein, the phrase “integer from X to Y” means any integer that includes the endpoints. For example, the phrase “integer from 1 to 5” means 1, 2, 3, 4, or 5. Additionally, the phrase “from X to Y” also includes the end points. For example, the phrase “from 1 to 5” includes the numerical values from 1 to 5 and includes the endpoints of 1 and 5. Unless context indicates otherwise, the phrase “between from X to Y” also includes the endpoints of X and Y.

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[0069] IPTS / 200163498.2 DOCKET NO. SES-020WO PATENT

[0070] As used herein, the phrase “ophthalmically acceptable” means having no persistent detrimental effect on the treated eye or the functioning thereof, or on the general health of the subject being treated. However, it will be recognized that transient effects such as minor irritation or a “stinging” sensation are common with topical ophthalmic administration of drugs and the existence of such transient effects is not inconsistent with the composition, formulation, or ingredient (e.g., excipient) in question being “ophthalmically acceptable” as herein defined. In some embodiments, the pharmaceutical compositions can be ophthalmically acceptable or suitable for ophthalmic administration.

[0071] In some embodiments, the term “therapeutic molecule” can be used interchangeably with “therapeutic compound,” “molecule,” or “therapeutic,” and refers to any polypeptide, or protein provided for herein.

[0072] As used herein, the term “position,” is meant to refer to a location in the sequence of a polypeptide. Positions may be numbered sequentially, or according to an established format, for example the EU numbering system based on Kabat' s amino acid positions of an antibody for Fc domain. For example, position 298 is an EU numbered position in the human antibody IgGl.

[0073] "Specific binding" or "specifically binds to" or is "specific for" a particular antigen, target, or an epitope means binding that is measurably different from a non-specific interaction. Specific binding can be measured, for example, by determining binding of a molecule compared to binding of a control molecule, which generally is a molecule of similar structure that does not have binding activity. For example, specific binding can be determined by competition with a control molecule that is similar to the target. For example, a protease can be specific for cleaving IgG or have preference for IgG over other types or classes of immunoglobulins, such as IgM or IgA.

[0074] As provided herein, the compounds and compositions provided for herein can be used in methods of treatment as provided herein. As used herein, the terms “treat,” “treated,” or “treating” mean both therapeutic treatment and prophylactic measures wherein the object is to slow down (lessen) an undesired physiological condition, disorder or disease, or obtain beneficial or desired clinical results. For purposes of these embodiments, beneficial or desired clinical results include, but are not limited to, alleviation of symptoms; diminishment of extent of condition, disorder or disease; stabilized (i.e., not worsening) state of condition, disorder or

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[0076] IPTS / 200163498.2 DOCKET NO. SES-020WO PATENT disease; delay in onset or slowing of condition, disorder or disease progression; amelioration of the condition, disorder or disease state or remission (whether partial or total), whether detectable or undetectable; an amelioration of at least one measurable physical parameter, not necessarily discernible by the patient; or enhancement or improvement of condition, disorder or disease. Treatment includes eliciting a clinically significant response without excessive levels of side effects. Treatment also includes prolonging survival, as applicable for a specific disease, as compared to expected survival if not receiving treatment. Thus, “treatment of an autoimmune condition” or “treating autoimmunity” means an activity that alleviates or ameliorates any of the primary phenomena or secondary symptoms associated with the autoimmune condition other condition described herein when the terms “treat,” “treated,” or “treating” are used in conjunction with such condition. For example, as provided for herein, the proteases provided for herein can be used to reduce immunoglobulins in a subject (patient) to treat an autoimmune disease or other pathology that is due to aberrant immunoglobulin expression or presence of immunoglobulins that are desired to be reduced in a patient for other reasons.

[0077] As used herein, terms “variant,” as it relates to a “molecule,” “therapeutic,” “therapeutic compound,” “compound,” “polypeptide,” or “protein” relate to the variants of such molecules, therapeutics, therapeutic compounds, compounds, polypeptides, and proteins disclosed herein.

[0078] Variant Fc Molecules

[0079] As used herein, "isotype" in reference to immunoglobulins refers to the immunoglobulin class (e.g., IgGl, IgG2, IgG3, IgG4, IgM, IgAl, IgA2, IgD, and IgE antibody) that is encoded by the heavy chain constant domain genes of such antibodies. The full-length amino acid sequence of each wild type human IgG constant region (including all domains, i.e., CHI domain, hinge, CH2 domain, and CH3 domain) is cataloged in the UniProt database available on-line, e.g., as P01857 (IgGl), P01859 (IgG2), P01860 (IgG3), and P01861 (IgG4), or different allotypes thereof (SEQ ID NOs: 1, 2, 3, and 4, respectively). The UniProt sequences are also hereby incorporated by reference in its entirety. As used herein, a domain of a heavy chain constant region, e.g., the hinge, is of an "IgGl isotype," "IgG2 isotype," "IgG3 isotype," or "IgG4 isotype," if the domain comprises the amino acid sequence of the corresponding domain of the respective isotype, or a variant thereof (that has a higher homology to the corresponding domain of the respective isotype than it does to that of the other isotypes).

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[0081] IPTS / 200163498.2 DOCKET NO. SES-020WO PATENT

[0082] “Allotype” refers to naturally occurring variants within a specific isotype group, which variants differ in a few amino acids (see, e.g., Jefferies et al. (2009) mAbs 1 : 1, which is hereby incorporated by reference in its entirety). Molecules described herein may be of any allotype.

[0083] A “wild-type” protein or portion thereof is a version of the protein as it is found in nature. An amino acid sequence of a wild-type protein, e.g., a heavy chain constant region, is the amino acid sequence of the protein as it occurs in nature. Due to allotypic differences, there can be more than one amino acid sequence for a wild-type protein. For example, there are several allotypes of naturally occurring human IgGl heavy chain constant regions (e g., Jeffries et al. (2009) mAbs 1 : 1).

[0084] As used herein, the term “chemical liabilities” is meant to refer to factors that affects a molecule’s immunogenicity. Thus, in some embodiments, “chemical liabilities” is meant to refer to, without limitation, post-translational modifications, aggregation, glycosylation, impurities, or formulation components. A “chemical liability” that affects immunogenicity is a liability that has increased immunogenicity and produces an undesired immune response in the subject. Modifications of the sequence can be made to reduce such liabilities. In some embodiments, the polypeptides provided herein exhibit reduced B-cell binding epitopes. In some embodiments, the polypeptides provided herein exhibit reduced T-cell epitopes. In some embodiments, the polypeptides provided herein exhibit reduced post-translational modifications. In some embodiments, the polypeptides provided herein exhibit increased expression in vitro. In some embodiments, the polypeptides provided herein exhibit increased pH stability. In some embodiments, the polypeptides provided herein exhibit reduced or no B-cell binding epitopes. In some embodiments, the polypeptides provided herein exhibit reduced or no T-cell epitopes. In some embodiments, the polypeptides provided herein exhibit reduced B-cell binding epitopes. In some embodiments, the polypeptides provided herein exhibit reduced T-cell epitopes. In some embodiments, the polypeptides provided herein exhibit no B-cell binding epitopes. In some embodiments, the polypeptides provided herein exhibit no T-cell epitopes. In some embodiments, the polypeptides provided herein exhibit reduced aggregation. In some embodiments, the polypeptides provided herein exhibit no post-translational modifications. In some embodiments, the polypeptides provided herein exhibit reduced B-cell binding epitopes; reduced T-cell epitopes; reduced post-translational modifications; increased expression in vitro; reduced aggregation; and / or increased pH stability.

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[0086] IPTS / 200163498.2 DOCKET NO. SES-020WO PATENT

[0087] An immunoglobulin may be from any of the commonly known isotypes, including but not limited to IgA, secretory IgA, IgG and IgM. The IgG isotype is divided in subclasses in certain species: IgGl, IgG2, IgG3 and IgG4 in humans, and IgGl, IgG2a, IgG2b and IgG3 in mice. In certain embodiments, the antibodies described herein are of the human IgGl or IgG2 subtype. Immunoglobulins, e.g., human IgGl, exist in several allotypes, which differ from each other in at most a few amino acids.

[0088] In some embodiments, the IgG proteins (hinge region underlined) are as provided in Table 1. An "Fc region" (fragment crystallizable region) or "Fc polypeptide" or "Fc" refers to the

[0089] C- terminal region of the heavy chain of an antibody that mediates the binding of the immunoglobulin to host tissues or factors, including binding to Fc receptors located on various cells of the immune system (e.g., effector cells) or to the first component (Clq) of the classical complement system. Thus, an Fc region of an antibody of isotype IgG comprises the heavy chain constant region of the antibody excluding the first constant region immunoglobulin domain (CHI). In IgG, IgA and IgD antibody isotypes, the Fc region comprises CH2 and CH3 constant domains in each of the antibody’s two heavy chains; IgM and IgE Fc regions comprise three

[0090] -15-

[0091] IPTS / 200163498.2 DOCKET NO. SES-020WO PATENT heavy chain constant domains (CH domains 2-4) in each polypeptide chain. For IgG, the Fc region comprises immunoglobulin domains consisting of the hinge, CH2 and CH3. For purposes herein, the Fc region is defined as starting at amino acid 216 and ending at amino acid 447, wherein the numbering is according to the EU index as in Kabat. Kabat et al. (1991) Sequences of Proteins of Immunological Interest, National Institutes of Health, Bethesda, MD, and according to FIGs.3c-3f of U.S. Pat. App. Pub. No.2008 / 0248028. The EU index, which can be referred to as EU numbering, is a well accepted nomenclature for referring to certain residues in the Fc region. Alternatively, there are linear equivalents where the first residue of the sequences of the Fc polypeptides can be referred to residue number 1 and so on and so forth. In some embodiments, the Fc region comprises the hinge region. The Fc may be a native (or naturally- occurring or wild-type) Fc, including any allotypic variant, or a variant Fc (e.g., a non- naturally occurring Fc), comprising, e g., 1, 2, 3, 4, 5, 1-5, 1-10 or 5-10 or more amino acid mutations, e.g., substitutions, additions or deletions. For example, a variant Fc may comprise an amino acid sequence that is at least 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% identical to a wildtype Fc. Modified or mutated Fes may have enhanced or reduced effector function and / or enhanced half-life. Fc may refer to this region in isolation or in the context of an Fc-comprising protein polypeptide such as a “binding protein comprising an Fc region,” also referred to as an “Fc fusion protein” (e.g., an antibody or immunoadhesin). In some embodiments, modified or variant Fc molecules have enhanced binding to FcyRIIb. The enhanced binding can be as compared to FcyRIIa, that is the Fc can bind preferentially to FcyRIIb as compared to FcyRIIa.

[0092] A "hinge", "hinge domain" or "hinge region" or "antibody hinge region" refers to the domain of a heavy chain constant region that joins the CHI domain to the CH2 domain and includes the upper, middle, and lower portions of the hinge (Roux et al. J. Immunol.1998 161 :4083). The hinge provides varying levels of flexibility between the binding and effector regions of an antibody and also provides sites for intermolecular disulfide bonding between the two heavy chain constant regions. The term “hinge” includes wild-type hinges (such as those set forth in Table 3), as well as variants thereof (e.g., non-naturally-occurring hinges or modified hinges). For example, the term “IgGl hinge” includes wild-type IgGl hinge, as shown below, and variants having 1, 2, 3, 4, 5, 1-3, 1-5, 3-5 and / or at most 5, 4, 3, 2, or 1 mutations, e.g., substitutions, deletions or additions. In some embodiments, the hinge regions are as provided in Table 2.

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[0094] IPTS / 200163498.2 DOCKET NO. SES-020WO PATENT

[0095] The term “CHI domain” refers to the heavy chain constant region linking the variable domain to the hinge in a heavy chain constant domain. As used herein, a CHI domain includes wild type CHI domains, as well as variants thereof (e.g., non-naturally-occurring CHI domains or modified CHI domains). For example, the term “CHI domain” includes wild-type CHI domains and variants thereof having 1, 2, 3, 4, 5, 1-3, 1-5, 3-5 and / or at most 5, 4, 3, 2, or 1 mutations, e.g., substitutions, deletions or additions.

[0096] The term “CH2 domain” refers to the heavy chain constant region linking the hinge to the CH3 domain in a heavy chain constant domain. As used herein, a CH2 domain includes wildtype CH2 domains, as well as variants thereof (e.g., non-naturally-occurring CH2 domains or modified CH2 domains). For example, the term “CH2 domain” includes wild-type CH2 domains and variants thereof having 1, 2, 3, 4, 5, 1-3, 1-5, 3-5 and / or at most 5, 4, 3, 2, or 1 mutations, e.g., substitutions, deletions or additions.

[0097] The term “CH3 domain” refers to the heavy chain constant region that is C-terminus to the CH2 domain in a heavy chain constant domain. As used herein, a CH3 domain includes wildtype CH3 domains, as well as variants thereof (e.g., non-naturally- occurring CH3 domains or modified CH3 domains). For example, the term “CH3 domain” includes wild-type CH3 domains and variants thereof having 1, 2, 3, 4, 5, 1-3, 1-5, 3-5 and / or at most 5, 4, 3, 2, or 1 mutations, e.g., substitutions, deletions or additions.

[0098] Provided herein are Fc polypeptides comprising Fc polypeptides, e.g., Fc polypeptides that have a mutated sequence region, relative to wild-type Fc polypeptide. Exemplary variant Fc molecules comprising Fc polypeptides include an IgGl hinge, a CHI domain, a CH2 domain and a CH3 domain, wherein at least one of these constant domains has residues that are not wild-type residues, as compared to SEQ ID NO: 1, 2, 3, or 4. A Fc polypeptide may have effector function

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[0100] IPTS / 200163498.2 DOCKET NO. SES-020WO PATENT similar to that of wild-type IgG, or may be engineered to have enhanced effector function relative to that of the wild-type IgG. A Fc polypeptide may comprise a wild-type CHI, hinge, CH2 and / or CH3 domain, or a variant thereof, e.g., a CHI, hinge, CH2 and / or CH3 domain having one or more amino acid substitutions, deletions or additions relative to the corresponding wild-type domain, and / or having an amino acid sequence that is at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical, or more, to the corresponding wild-type sequence.

[0101] In some embodiments, the IgG proteins are as provided in Table 6.

[0102] In some embodiments, a Fc polypeptide comprises one or more mutations that confers resistance to recognition by a protease. In some embodiments, a Fc polypeptide comprises one or more mutations that confers resistance to proteolytic cleavage. In some embodiments, a Fc polypeptide comprises one or more mutations that confer resistance to recognition by a protease and proteolytic cleavage.

[0103] The numbering of the residues of the Fc polypeptides provided for herein, unless otherwise noted, is according to the EU index or EU numbering.

[0104] In some embodiments, a Fc polypeptide comprises an amino acid sequence having at least 50%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%, or 100% sequence identity to SEQ ID NO: 1, provided that the Fc polypeptide comprises an amino acid mutation at any one, or more, position between positions 234 and 329, as compared to SEQ ID NO: 1, wherein the mutation comprises an insertion, a deletion or a substitution.

[0105] In some embodiments, a Fc polypeptide comprises an amino acid sequence having at least 50%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at

[0106] -18-

[0107] IPTS / 200163498.2 DOCKET NO. SES-020WO PATENT least 99%, or 100% sequence identity to SEQ ID NO: 1 , provided that the Fc polypeptide comprises a mutation corresponding to L234A mutation, as compared to SEQ ID NO: 1.

[0108] In some embodiments, a Fc polypeptide comprises an amino acid sequence having at least 50%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%, or 100% sequence identity to SEQ ID NO: 1, provided that the Fc polypeptide comprises a mutation corresponding to L235A mutation, as compared to SEQ ID NO: 1.

[0109] In some embodiments, a Fc polypeptide comprises an amino acid sequence having at least 50%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%, or 100% sequence identity to SEQ ID NO: 1, provided that the Fc polypeptide comprises a mutation corresponding to G237A mutation, as compared to SEQ ID NO: 1.

[0110] In some embodiments, a Fc polypeptide comprises an amino acid sequence having at least 50%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%, or 100% sequence identity to SEQ ID NO: 1, provided that the Fc polypeptide comprises a mutation corresponding to Y296Q mutation, as compared to SEQ ID NO: 1.

[0111] In some embodiments, a Fc polypeptide comprises an amino acid sequence having at least 50%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%, or 100% sequence identity to SEQ ID NO: 1, provided that the Fc polypeptide comprises a mutation corresponding to P329K mutation, as compared to SEQ ID NO: 1.

[0112] In some embodiments, a Fc polypeptide comprises an amino acid sequence having at least 50%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%, or 100% sequence identity to SEQ ID NO: 1, provided that the Fc polypeptide comprises a set of mutations corresponding to L234A, L235A, and G237A mutations, as compared to SEQ ID NO: 1.

[0113] In some embodiments, a Fc polypeptide comprises an amino acid sequence having at least 50%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at

[0114] -19-

[0115] IPTS / 200163498.2 DOCKET NO. SES-020WO PATENT least 99%, or 100% sequence identity to SEQ ID NO: 1, provided that the Fc polypeptide comprises a set of mutations corresponding to Y296Q and P329K mutations, as compared to SEQ ID NO: 1.

[0116] In some embodiments, a Fc polypeptide comprises an amino acid sequence having at least 50%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%, or 100% sequence identity to SEQ ID NO: 1, provided that the Fc polypeptide comprises a set of mutations corresponding to L234A, L235A, G237A, Y296Q, and P329K mutations, as compared to SEQ ID NO: 1.

[0117] In some embodiments, the variant Fc polypeptide comprises at least one mutation that extends the half-life of the Fc polypeptide. In some embodiments, the at least one mutation that extends the half-life of the Fc polypeptide, is such as those known in the art and may be found in U.S. Patent Nos. 7,658,921, 11,059,892, or International Application No. PCT / US2008 / 088053, each of which is hereby incorporated by reference in its entirety. In some embodiments, the at least one mutation that extends the half-life of the Fc polypeptide, is such as those known in the art, such as, without limitation, a set of mutations of M428L and N434S (“LS” mutations), a set of mutations of M252Y, S254T, and T256E (“YTE” mutations), a set of mutations of L309D, Q311H, and N434Y, a set of mutations of L309D, Q311H, and N434S, a set of mutations of V309D, Q311H, and N434Y, or a set of mutations of V309D, Q311H, and N434S. The extension mutations can be combined with or used independently of the other Fc mutations provided for herein.

[0118] In some embodiments, a variant Fc polypeptide comprising at least one mutation that extends the half-life of the Fc polypeptide comprises an amino acid sequence having at least 50%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%, or 100% sequence identity to any one of SEQ ID NO: 11, 12, and 13. In some embodiments, a variant Fc polypeptide comprising at least one mutation that extends the half-life of the Fc polypeptide comprises an amino acid sequence having at least 50%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%, or 100% sequence identity to SEQ ID NO: 11. In some embodiments, a variant Fc polypeptide comprising

[0119] -20-

[0120] IPTS / 200163498.2 DOCKET NO. SES-020WO PATENT at least one mutation that extends the half-life of the Fc polypeptide comprises an amino acid sequence having at least 50%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%, or 100% sequence identity to SEQ ID NO: 12. In some embodiments, a variant Fc polypeptide comprising at least one mutation that extends the half-life of the Fc polypeptide comprises an amino acid sequence having at least 50%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%, or 100% sequence identity to SEQ ID NO: 13.

[0121] In some embodiments, a variant Fc polypeptide comprising at least one mutation that extends the half-life of the Fc polypeptide comprises an amino acid sequence selected from any one of SEQ ID NO: 11, 12, and 13. In some embodiments, a variant Fc polypeptide comprising at least one mutation that extends the half-life of the Fc polypeptide comprises an amino acid sequence of SEQ ID NO: 11. In some embodiments, a variant Fc polypeptide comprising at least one mutation that extends the half-life of the Fc polypeptide comprises an amino acid sequence of SEQ ID NO: 12. In some embodiments, a variant Fc polypeptide comprising at least one mutation that extends the half-life of the Fc polypeptide comprises an amino acid sequence of SEQ ID NO: 13.

[0122] In some embodiments, a variant Fc polypeptide comprises an amino acid sequence having at least 50%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%, or 100% sequence identity to SEQ ID NO: 1, provided that the variant Fc polypeptide comprises a set of mutations corresponding to Y296Q, M428L and N434S mutations, as compared to SEQ ID NO: 1.

[0123] In some embodiments, a variant Fc polypeptide comprises an amino acid sequence having at least 50%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%, or 100% sequence identity to SEQ ID NO: 1, provided that the variant Fc polypeptide comprises a set of mutations corresponding to Y296Q, M252Y, S254T, and T256E mutations, as compared to SEQ ID NO: 1.

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[0125] IPTS / 200163498.2 DOCKET NO. SES-020WO PATENT

[0126] In some embodiments, a variant Fc polypeptide comprises an amino acid sequence having at least 50%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%, or 100% sequence identity to SEQ ID NO: 1, provided that the variant Fc polypeptide comprises a set of mutations corresponding to Y296Q, L309D, Q311H, and N434Y mutations, as compared to SEQ ID NO: 1.

[0127] In some embodiments, a variant Fc polypeptide comprises an amino acid sequence having at least 50%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%, or 100% sequence identity to SEQ ID NO: 1, provided that the variant Fc polypeptide comprises a set of mutations corresponding to Y296Q, L309D, Q311H, and N434S mutations, as compared to SEQ ID NO: 1.

[0128] In some embodiments, a variant Fc polypeptide comprises an amino acid sequence having at least 50%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%, or 100% sequence identity to SEQ ID NO: 1, provided that the variant Fc polypeptide comprises a set of mutations corresponding to Y296Q, V309D, Q31 1H, and N434Y mutations, as compared to SEQ ID NO: 1.

[0129] In some embodiments, a variant Fc polypeptide comprises an amino acid sequence having at least 50%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%, or 100% sequence identity to SEQ ID NO: 1, provided that the variant Fc polypeptide comprises a set of mutations corresponding to Y296Q, V309D, Q31 1H, and N434S mutations, as compared to SEQ ID NO: 1.

[0130] In some embodiments, a variant Fc polypeptide comprises an amino acid sequence having at least 50%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%, or 100% sequence identity to SEQ ID NO: 1, provided that the variant Fc polypeptide comprises a set of mutations corresponding to P329K, M428L and N434S mutations, as compared to SEQ ID NO: 1.

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[0132] IPTS / 200163498.2 DOCKET NO. SES-020WO PATENT

[0133] In some embodiments, a variant Fc polypeptide comprises an amino acid sequence having at least 50%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%, or 100% sequence identity to SEQ ID NO: 1, provided that the variant Fc polypeptide comprises a set of mutations corresponding to P329K, M252Y, S254T, and T256E mutations, as compared to SEQ ID NO: 1.

[0134] In some embodiments, a variant Fc polypeptide comprises an amino acid sequence having at least 50%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%, or 100% sequence identity to SEQ ID NO: 1, provided that the variant Fc polypeptide comprises a set of mutations corresponding to P329K, L309D, Q311H, and N434Y mutations, as compared to SEQ ID NO: 1.

[0135] In some embodiments, a variant Fc polypeptide comprises an amino acid sequence having at least 50%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%, or 100% sequence identity to SEQ ID NO: 1, provided that the variant Fc polypeptide comprises a set of mutations corresponding to P329K, L309D, Q311H, and N434S mutations, as compared to SEQ ID NO: 1.

[0136] In some embodiments, a variant Fc polypeptide comprises an amino acid sequence having at least 50%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%, or 100% sequence identity to SEQ ID NO: 1, provided that the variant Fc polypeptide comprises a set of mutations corresponding to P329K, V309D, Q311H, and N434Y mutations, as compared to SEQ ID NO: 1.

[0137] In some embodiments, a variant Fc polypeptide comprises an amino acid sequence having at least 50%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%, or 100% sequence identity to SEQ ID NO: 1, provided that the variant Fc polypeptide comprises a set of mutations corresponding to P329K, V309D, Q311H, and N434S mutations, as compared to SEQ ID NO: 1.

[0138] -23-

[0139] IPTS / 200163498.2 DOCKET NO. SES-020WO PATENT

[0140] In some embodiments, a variant Fc polypeptide comprises an amino acid sequence having at least 50%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%, or 100% sequence identity to SEQ ID NO: 1, provided that the variant Fc polypeptide comprises a set of mutations corresponding to Y296Q, P329K, M428L and N434S mutations, as compared to SEQ ID NO: 1.

[0141] In some embodiments, a variant Fc polypeptide comprises an amino acid sequence having at least 50%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%, or 100% sequence identity to SEQ ID NO: 1, provided that the variant Fc polypeptide comprises a set of mutations corresponding to Y296Q, P329K, M252Y, S254T, and T256E mutations, as compared to SEQ ID NO: 1.

[0142] In some embodiments, a variant Fc polypeptide comprises an amino acid sequence having at least 50%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%, or 100% sequence identity to SEQ ID NO: 1, provided that the variant Fc polypeptide comprises a set of mutations corresponding to Y296Q, P329K, L309D, Q311H, and N434Y mutations, as compared to SEQ ID NO: 1.

[0143] In some embodiments, a variant Fc polypeptide comprises an amino acid sequence having at least 50%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%, or 100% sequence identity to SEQ ID NO: 1, provided that the variant Fc polypeptide comprises a set of mutations corresponding to Y296Q, P329K, L309D, Q311H, and N434S mutations, as compared to SEQ ID NO: 1.

[0144] In some embodiments, a variant Fc polypeptide comprises an amino acid sequence having at least 50%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%, or 100% sequence identity to SEQ ID NO: 1, provided that the variant Fc polypeptide comprises a set of mutations corresponding to Y296Q, P329K, V309D, Q311H, and N434Y mutations, as compared to SEQ ID NO: 1.

[0145] -24-

[0146] IPTS / 200163498.2 DOCKET NO. SES-020WO PATENT

[0147] In some embodiments, a variant Fc polypeptide comprises an amino acid sequence having at least 50%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%, or 100% sequence identity to SEQ ID NO: 1, provided that the variant Fc polypeptide comprises a set of mutations corresponding to Y296Q, P329K, V309D, Q311H, and N434S mutations, as compared to SEQ ID NO: 1.

[0148] In some embodiments, a variant Fc polypeptide comprises an amino acid sequence having at least 50%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%, or 100% sequence identity to SEQ ID NO: 1, provided that the variant Fc polypeptide comprises a set of mutations corresponding to L234A, L235A, G237A, Y296Q, P329K, M428L and N434S mutations, as compared to SEQ ID NO: 1.

[0149] In some embodiments, a variant Fc polypeptide comprises an amino acid sequence having at least 50%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%, or 100% sequence identity to SEQ ID NO: 1, provided that the variant Fc polypeptide comprises a set of mutations corresponding to L234A, L235A, G237A, Y296Q, P329K, M252Y, S254T, and T256E mutations, as compared to SEQ ID NO: 1.

[0150] In some embodiments, a variant Fc polypeptide comprises an amino acid sequence having at least 50%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%, or 100% sequence identity to SEQ ID NO: 1, provided that the variant Fc polypeptide comprises a set of mutations corresponding to L234A, L235A, G237A, Y296Q, P329K, L309D, Q311H, and N434Y mutations, as compared to SEQ ID NO: 1.

[0151] In some embodiments, a variant Fc polypeptide comprises an amino acid sequence having at least 50%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%, or 100% sequence identity to SEQ ID NO: 1, provided that the variant Fc polypeptide comprises a set of mutations corresponding to L234A, L235A, G237A, Y296Q, P329K, L309D, Q311H, and N434S mutations, as compared to SEQ ID NO: 1 .

[0152] -25-

[0153] IPTS / 200163498.2 DOCKET NO. SES-020WO PATENT

[0154] In some embodiments, a variant Fc polypeptide comprises an amino acid sequence having at least 50%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%, or 100% sequence identity to SEQ ID NO: 1, provided that the variant Fc polypeptide comprises a set of mutations corresponding to L234A, L235A, G237A, Y296Q, P329K, V309D, Q311H, and N434Y mutations, as compared to SEQ ID NO: 1 .

[0155] In some embodiments, a variant Fc polypeptide comprises an amino acid sequence having at least 50%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%, or 100% sequence identity to SEQ ID NO: 1, provided that the variant Fc polypeptide comprises a set of mutations corresponding to L234A, L235A, G237A, Y296Q, P329K, V309D, Q311H, and N434S mutations, as compared to SEQ ID NO: 1 .

[0156] In some embodiments, a variant Fc polypeptide comprises an amino acid sequence having at least 50%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%, or 100% sequence identity to SEQ ID NO: 10, provided that the variant Fc polypeptide comprises a set of mutations corresponding to Y296Q, M428L and N434S mutations, according to EU numbering.

[0157] In some embodiments, a variant Fc polypeptide comprises an amino acid sequence having at least 50%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%, or 100% sequence identity to SEQ ID NO: 10, provided that the variant Fc polypeptide comprises a set of mutations corresponding to Y296Q, M252Y, S254T, and T256E mutations, according to EU numbering.

[0158] In some embodiments, a variant Fc polypeptide comprises an amino acid sequence having at least 50%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%, or 100% sequence identity to SEQ ID NO: 10, provided that the variant Fc polypeptide comprises a set of mutations corresponding to Y296Q, L309D, Q311H, and N434Y mutations, according to EU numbering.

[0159] -26-

[0160] IPTS / 200163498.2 DOCKET NO. SES-020WO PATENT

[0161] In some embodiments, a variant Fc polypeptide comprises an amino acid sequence having at least 50%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%, or 100% sequence identity to SEQ ID NO: 10, provided that the variant Fc polypeptide comprises a set of mutations corresponding to Y296Q, L309D, Q311H, and N434S mutations, according to EU numbering.

[0162] In some embodiments, a variant Fc polypeptide comprises an amino acid sequence having at least 50%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%, or 100% sequence identity to SEQ ID NO: 10, provided that the variant Fc polypeptide comprises a set of mutations corresponding to Y296Q, V309D, Q311H, and N434Y mutations, according to EU numbering.

[0163] In some embodiments, a variant Fc polypeptide comprises an amino acid sequence having at least 50%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%, or 100% sequence identity to SEQ ID NO: 10, provided that the variant Fc polypeptide comprises a set of mutations corresponding to Y296Q, V309D, Q31 1H, and N434S mutations, according to EU numbering.

[0164] In some embodiments, a variant Fc polypeptide comprises an amino acid sequence having at least 50%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%, or 100% sequence identity to SEQ ID NO: 10, provided that the variant Fc polypeptide comprises a set of mutations corresponding to P329K, M428L and N434S mutations, according to EU numbering.

[0165] In some embodiments, a variant Fc polypeptide comprises an amino acid sequence having at least 50%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%, or 100% sequence identity to SEQ ID NO: 10, provided that the variant Fc polypeptide comprises a set of mutations corresponding to P329K, M252Y, S254T, and T256E mutations, according to EU numbering.

[0166] -27-

[0167] IPTS / 200163498.2 DOCKET NO. SES-020WO PATENT

[0168] In some embodiments, a variant Fc polypeptide comprises an amino acid sequence having at least 50%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%, or 100% sequence identity to SEQ ID NO: 10, provided that the variant Fc polypeptide comprises a set of mutations corresponding to P329K, L309D, Q311H, and N434Y mutations, according to EU numbering.

[0169] In some embodiments, a variant Fc polypeptide comprises an amino acid sequence having at least 50%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%, or 100% sequence identity to SEQ ID NO: 10, provided that the variant Fc polypeptide comprises a set of mutations corresponding to P329K, L309D, Q311H, and N434S mutations, according to EU numbering.

[0170] In some embodiments, a variant Fc polypeptide comprises an amino acid sequence having at least 50%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%, or 100% sequence identity to SEQ ID NO: 10, provided that the variant Fc polypeptide comprises a set of mutations corresponding to P329K, V309D, Q311H, and N434Y mutations, according to EU numbering.

[0171] In some embodiments, a variant Fc polypeptide comprises an amino acid sequence having at least 50%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%, or 100% sequence identity to SEQ ID NO: 10, provided that the variant Fc polypeptide comprises a set of mutations corresponding to P329K, V309D, Q311H, and N434S mutations, according to EU numbering.

[0172] In some embodiments, a variant Fc polypeptide comprises an amino acid sequence having at least 50%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%, or 100% sequence identity to SEQ ID NO: 10, provided that the variant Fc polypeptide comprises a set of mutations corresponding to Y296Q, P329K, M428L and N434S mutations, according to EU numbering.

[0173] -28-

[0174] IPTS / 200163498.2 DOCKET NO. SES-020WO PATENT

[0175] In some embodiments, a variant Fc polypeptide comprises an amino acid sequence having at least 50%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%, or 100% sequence identity to SEQ ID NO: 10, provided that the variant Fc polypeptide comprises a set of mutations corresponding to Y296Q, P329K, M252Y, S254T, and T256E mutations, according to EU numbering.

[0176] In some embodiments, a variant Fc polypeptide comprises an amino acid sequence having at least 50%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%, or 100% sequence identity to SEQ ID NO: 10, provided that the variant Fc polypeptide comprises a set of mutations corresponding to Y296Q, P329K, L309D, Q311H, and N434Y mutations, according to EU numbering.

[0177] In some embodiments, a variant Fc polypeptide comprises an amino acid sequence having at least 50%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%, or 100% sequence identity to SEQ ID NO: 10, provided that the variant Fc polypeptide comprises a set of mutations corresponding to Y296Q, P329K, L309D, Q311H, and N434S mutations, according to EU numbering.

[0178] In some embodiments, a variant Fc polypeptide comprises an amino acid sequence having at least 50%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%, or 100% sequence identity to SEQ ID NO: 10, provided that the variant Fc polypeptide comprises a set of mutations corresponding to Y296Q, P329K, V309D, Q311H, and N434Y mutations, according to EU numbering.

[0179] In some embodiments, a variant Fc polypeptide comprises an amino acid sequence having at least 50%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%, or 100% sequence identity to SEQ ID NO: 10, provided that the variant Fc polypeptide comprises a set of mutations corresponding to Y296Q, P329K, V309D, Q311H, and N434S mutations, according to EU numbering.

[0180] IPTS / 200163498.2 DOCKET NO. SES-020WO PATENT

[0181] In some embodiments, a variant Fc polypeptide comprises an amino acid sequence having at least 50%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%, or 100% sequence identity to SEQ ID NO: 10, provided that the variant Fc polypeptide comprises a set of mutations corresponding to L234A, L235A, G237A, Y296Q, P329K, M428L and N434S mutations, according to EU numbering.

[0182] In some embodiments, a variant Fc polypeptide comprises an amino acid sequence having at least 50%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%, or 100% sequence identity to SEQ ID NO: 10, provided that the variant Fc polypeptide comprises a set of mutations corresponding to L234A, L235A, G237A, Y296Q, P329K, M252Y, S254T, and T256E mutations, according to EU numbering.

[0183] In some embodiments, a variant Fc polypeptide comprises an amino acid sequence having at least 50%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%, or 100% sequence identity to SEQ ID NO: 10, provided that the variant Fc polypeptide comprises a set of mutations corresponding to L234A, L235A, G237A, Y296Q, P329K, L309D, Q311H, and N434Y mutations, according to EU numbering.

[0184] In some embodiments, a variant Fc polypeptide comprises an amino acid sequence having at least 50%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%, or 100% sequence identity to SEQ ID NO: 10, provided that the variant Fc polypeptide comprises a set of mutations corresponding to L234A, L235A, G237A, Y296Q, P329K, L309D, Q311H, and N434S mutations, according to EU numbering.

[0185] In some embodiments, a variant Fc polypeptide comprises an amino acid sequence having at least 50%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%, or 100% sequence identity to SEQ ID NO: 10, provided that the variant Fc polypeptide comprises a set of mutations corresponding to L234A, L235A, G237A, Y296Q, P329K, V309D, Q311H, and N434Y mutations, according to EU numbering.

[0186] -30-

[0187] IPTS / 200163498.2 DOCKET NO. SES-020WO PATENT

[0188] In some embodiments, a variant Fc polypeptide comprises an amino acid sequence having at least 50%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%, or 100% sequence identity to SEQ ID NO: 10, provided that the variant Fc polypeptide comprises a set of mutations corresponding to L234A, L235A, G237A, Y296Q, P329K, V309D, Q311H, and N434S mutations, according to EU numbering.

[0189] In some embodiments, a variant Fc polypeptide comprises an amino acid sequence having at least 50%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%, or 100% sequence identity to SEQ ID NO: 19, 20, 21, 22, 23, 24, or 25, provided that the variant Fc polypeptide comprises a set of mutations corresponding to Y296Q, M428L and N434S mutations, according to EU numbering.

[0190] In some embodiments, a variant Fc polypeptide comprises an amino acid sequence having at least 50%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%, or 100% sequence identity to SEQ ID NO: 19, 20, 21, 22, 23, 24, or 25, provided that the variant Fc polypeptide comprises a set of mutations corresponding to Y296Q, M252Y, S254T, and T256E mutations, according to EU numbering.

[0191] In some embodiments, a variant Fc polypeptide comprises an amino acid sequence having at least 50%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%, or 100% sequence identity to SEQ ID NO: 19, 20, 21, 22, 23, 24, or 25, provided that the variant Fc polypeptide comprises a set of mutations corresponding to Y296Q, L309D, Q31 1H, and N434Y mutations, according to EU numbering.

[0192] In some embodiments, a variant Fc polypeptide comprises an amino acid sequence having at least 50%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%, or 100% sequence identity to SEQ ID NO: 19, 20, 21, 22, 23, 24, or 25, provided that the variant Fc polypeptide comprises a set of mutations corresponding to Y296Q, L309D, Q31 1H, and N434S mutations, according to EU numbering.

[0193] -31-

[0194] IPTS / 200163498.2 DOCKET NO. SES-020WO PATENT

[0195] In some embodiments, a variant Fc polypeptide comprises an amino acid sequence having at least 50%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%, or 100% sequence identity to SEQ ID NO: 19, 20, 21, 22, 23, 24, or 25, provided that the variant Fc polypeptide comprises a set of mutations corresponding to Y296Q, V309D, Q311H, and N434Y mutations, according to EU numbering.

[0196] In some embodiments, a variant Fc polypeptide comprises an amino acid sequence having at least 50%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%, or 100% sequence identity to SEQ ID NO: 19, 20, 21, 22, 23, 24, or 25, provided that the variant Fc polypeptide comprises a set of mutations corresponding to Y296Q, V309D, Q31 1H, and N434S mutations, according to EU numbering.

[0197] In some embodiments, a variant Fc polypeptide comprises an amino acid sequence having at least 50%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%, or 100% sequence identity to SEQ ID NO: 19, 20, 21, 22, 23, 24, or 25, provided that the variant Fc polypeptide comprises a set of mutations corresponding to P329K, M428L and N434S mutations, according to EU numbering.

[0198] In some embodiments, a variant Fc polypeptide comprises an amino acid sequence having at least 50%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%, or 100% sequence identity to SEQ ID NO: 19, 20, 21, 22, 23, 24, or 25, provided that the variant Fc polypeptide comprises a set of mutations corresponding to P329K, M252Y, S254T, and T256E mutations, according to EU numbering.

[0199] In some embodiments, a variant Fc polypeptide comprises an amino acid sequence having at least 50%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%, or 100% sequence identity to SEQ ID NO: 19, 20, 21, 22, 23, 24, or 25, provided that the variant Fc polypeptide comprises a set of mutations corresponding to P329K, L309D, Q31 1H, and N434Y mutations, according to EU numbering.

[0200] -32-

[0201] IPTS / 200163498.2 DOCKET NO. SES-020WO PATENT

[0202] In some embodiments, a variant Fc polypeptide comprises an amino acid sequence having at least 50%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%, or 100% sequence identity to SEQ ID NO: 19, 20, 21, 22, 23, 24, or 25, provided that the variant Fc polypeptide comprises a set of mutations corresponding to P329K, L309D, Q31 1H, and N434S mutations, according to EU numbering.

[0203] In some embodiments, a variant Fc polypeptide comprises an amino acid sequence having at least 50%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%, or 100% sequence identity to SEQ ID NO: 19, 20, 21, 22, 23, 24, or 25, provided that the variant Fc polypeptide comprises a set of mutations corresponding to P329K, V309D, Q31 1H, and N434Y mutations, according to EU numbering.

[0204] In some embodiments, a variant Fc polypeptide comprises an amino acid sequence having at least 50%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%, or 100% sequence identity to SEQ ID NO: 19, 20, 21, 22, 23, 24, or 25, provided that the variant Fc polypeptide comprises a set of mutations corresponding to P329K, V309D, Q311H, and N434S mutations, according to EU numbering.

[0205] In some embodiments, a variant Fc polypeptide comprises an amino acid sequence having at least 50%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%, or 100% sequence identity to SEQ ID NO: 19, 20, 21, 22, 23, 24, or 25, provided that the variant Fc polypeptide comprises a set of mutations corresponding to Y296Q, P329K, M428L and N434S mutations, according to EU numbering.

[0206] In some embodiments, a variant Fc polypeptide comprises an amino acid sequence having at least 50%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%, or 100% sequence identity to SEQ ID NO: 19, 20, 21, 22, 23, 24, or 25, provided that the variant Fc polypeptide comprises a set of mutations corresponding to Y296Q, P329K, M252Y, S254T, and T256E mutations, according to EU numbering.

[0207] -33-

[0208] IPTS / 200163498.2 DOCKET NO. SES-020WO PATENT

[0209] In some embodiments, a variant Fc polypeptide comprises an amino acid sequence having at least 50%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%, or 100% sequence identity to SEQ ID NO: 19, 20, 21, 22, 23, 24, or 25, provided that the variant Fc polypeptide comprises a set of mutations corresponding to Y296Q, P329K, L309D, Q31 1H, and N434Y mutations, according to EU numbering.

[0210] In some embodiments, a variant Fc polypeptide comprises an amino acid sequence having at least 50%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%, or 100% sequence identity to SEQ ID NO: 19, 20, 21, 22, 23, 24, or 25, provided that the variant Fc polypeptide comprises a set of mutations corresponding to Y296Q, P329K, L309D, Q311H, and N434S mutations, according to EU numbering.

[0211] In some embodiments, a variant Fc polypeptide comprises an amino acid sequence having at least 50%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%, or 100% sequence identity to SEQ ID NO: 19, 20, 21, 22, 23, 24, or 25, provided that the variant Fc polypeptide comprises a set of mutations corresponding to Y296Q, P329K, V309D, Q311H, and N434Y mutations, according to EU numbering.

[0212] In some embodiments, a variant Fc polypeptide comprises an amino acid sequence having at least 50%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%, or 100% sequence identity to SEQ ID NO: 19, 20, 21, 22, 23, 24, or 25, provided that the variant Fc polypeptide comprises a set of mutations corresponding to Y296Q, P329K, V309D, Q311H, and N434S mutations, according to EU numbering.

[0213] In some embodiments, a variant Fc polypeptide comprises an amino acid sequence having at least 50%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%, or 100% sequence identity to SEQ ID NO: 19, 20, 21, 22, 23, 24, or 25, provided that the variant Fc polypeptide comprises a set of mutations corresponding to L234A, L235A, G237A, Y296Q, P329K, M428L and N434S mutations, according to EU numbering.

[0214] -34-

[0215] IPTS / 200163498.2 DOCKET NO. SES-020WO PATENT

[0216] In some embodiments, a variant Fc polypeptide comprises an amino acid sequence having at least 50%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%, or 100% sequence identity to SEQ ID NO: 19, 20, 21, 22, 23, 24, or 25, provided that the variant Fc polypeptide comprises a set of mutations corresponding to L234A, L235A, G237A, Y296Q, P329K, M252Y, S254T, and T256E mutations, according to EU numbering.

[0217] In some embodiments, a variant Fc polypeptide comprises an amino acid sequence having at least 50%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%, or 100% sequence identity to SEQ ID NO: 19, 20, 21, 22, 23, 24, or 25, provided that the variant Fc polypeptide comprises a set of mutations corresponding to L234A, L235A, G237A, Y296Q, P329K, L309D, Q311H, and N434Y mutations, according to EU numbering.

[0218] In some embodiments, a variant Fc polypeptide comprises an amino acid sequence having at least 50%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%, or 100% sequence identity to SEQ ID NO: 19, 20, 21, 22, 23, 24, or 25, provided that the variant Fc polypeptide comprises a set of mutations corresponding to L234A, L235A, G237A, Y296Q, P329K, L309D, Q311H, and N434S mutations, according to EU numbering.

[0219] In some embodiments, a variant Fc polypeptide comprises an amino acid sequence having at least 50%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%, or 100% sequence identity to SEQ ID NO: 19, 20, 21, 22, 23, 24, or 25, provided that the variant Fc polypeptide comprises a set of mutations corresponding to L234A, L235A, G237A, Y296Q, P329K, V309D, Q311H, and N434Y mutations, according to EU numbering.

[0220] In some embodiments, a variant Fc polypeptide comprises an amino acid sequence having at least 50%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least

[0221] -35-

[0222] IPTS / 200163498.2 DOCKET NO. SES-020WO PATENT

[0223] 98%, or at least 99%, or 100% sequence identity to SEQ ID NO: 19, 20, 21 , 22, 23, 24, or 25, provided that the variant Fc polypeptide comprises a set of mutations corresponding to L234A, L235A, G237A, Y296Q, P329K, V309D, Q311H, and N434S mutations, according to EU numbering.

[0224] In some embodiments, a variant Fc polypeptide comprises an amino acid sequence having at least 50%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%, or 100% sequence identity to SEQ ID NO: 23, provided that the variant Fc polypeptide comprises a set of mutations corresponding to Y296Q, M428L and N434S mutations, according to EU numbering.

[0225] In some embodiments, a variant Fc polypeptide comprises an amino acid sequence having at least 50%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%, or 100% sequence identity to SEQ ID NO: 23, provided that the variant Fc polypeptide comprises a set of mutations corresponding to Y296Q, M252Y, S254T, and T256E mutations, according to EU numbering.

[0226] In some embodiments, a variant Fc polypeptide comprises an amino acid sequence having at least 50%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%, or 100% sequence identity to SEQ ID NO: 23, provided that the variant Fc polypeptide comprises a set of mutations corresponding to Y296Q, L309D, Q311H, and N434Y mutations, according to EU numbering.

[0227] In some embodiments, a variant Fc polypeptide comprises an amino acid sequence having at least 50%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%, or 100% sequence identity to SEQ ID NO: 23, provided that the variant Fc polypeptide comprises a set of mutations corresponding to Y296Q, L309D, Q311H, and N434S mutations, according to EU numbering.

[0228] In some embodiments, a variant Fc polypeptide comprises an amino acid sequence having at least 50%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least

[0229] -36-

[0230] IPTS / 200163498.2 DOCKET NO. SES-020WO PATENT

[0231] 98%, or at least 99%, or 100% sequence identity to SEQ ID NO: 23, provided that the variant Fc polypeptide comprises a set of mutations corresponding to Y296Q, V309D, Q311H, and N434Y mutations, according to EU numbering.

[0232] In some embodiments, a variant Fc polypeptide comprises an amino acid sequence having at least 50%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%, or 100% sequence identity to SEQ ID NO: 23, provided that the variant Fc polypeptide comprises a set of mutations corresponding to Y296Q, V309D, Q311H, and N434S mutations, according to EU numbering.

[0233] In some embodiments, a variant Fc polypeptide comprises an amino acid sequence having at least 50%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%, or 100% sequence identity to SEQ ID NO: 23, provided that the variant Fc polypeptide comprises a set of mutations corresponding to P329K, M428L and N434S mutations, according to EU numbering.

[0234] In some embodiments, a variant Fc polypeptide comprises an amino acid sequence having at least 50%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%, or 100% sequence identity to SEQ ID NO: 23, provided that the variant Fc polypeptide comprises a set of mutations corresponding to P329K, M252Y, S254T, and T256E mutations, according to EU numbering.

[0235] In some embodiments, a variant Fc polypeptide comprises an amino acid sequence having at least 50%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%, or 100% sequence identity to SEQ ID NO: 23, provided that the variant Fc polypeptide comprises a set of mutations corresponding to P329K, L309D, Q311H, and N434Y mutations, according to EU numbering.

[0236] In some embodiments, a variant Fc polypeptide comprises an amino acid sequence having at least 50%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%, or 100% sequence identity to SEQ ID NO: 23, provided that the variant Fc

[0237] IPTS / 200163498.2 DOCKET NO. SES-020WO PATENT polypeptide comprises a set of mutations corresponding to P329K, L309D, Q31 1H, and N434S mutations, according to EU numbering.

[0238] In some embodiments, a variant Fc polypeptide comprises an amino acid sequence having at least 50%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%, or 100% sequence identity to SEQ ID NO: 23, provided that the variant Fc polypeptide comprises a set of mutations corresponding to P329K, V309D, Q311H, and N434Y mutations, according to EU numbering.

[0239] In some embodiments, a variant Fc polypeptide comprises an amino acid sequence having at least 50%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%, or 100% sequence identity to SEQ ID NO: 23, provided that the variant Fc polypeptide comprises a set of mutations corresponding to P329K, V309D, Q311H, and N434S mutations, according to EU numbering.

[0240] In some embodiments, a variant Fc polypeptide comprises an amino acid sequence having at least 50%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%, or 100% sequence identity to SEQ ID NO: 23, provided that the variant Fc polypeptide comprises a set of mutations corresponding to Y296Q, P329K, M428L and N434S mutations, according to EU numbering.

[0241] In some embodiments, a variant Fc polypeptide comprises an amino acid sequence having at least 50%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%, or 100% sequence identity to SEQ ID NO: 23, provided that the variant Fc polypeptide comprises a set of mutations corresponding to Y296Q, P329K, M252Y, S254T, and T256E mutations, according to EU numbering.

[0242] In some embodiments, a variant Fc polypeptide comprises an amino acid sequence having at least 50%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%, or 100% sequence identity to SEQ ID NO: 23, provided that the variant Fc

[0243] -38-

[0244] IPTS / 200163498.2 DOCKET NO. SES-020WO PATENT polypeptide comprises a set of mutations corresponding to Y296Q, P329K, L309D, Q31 1H, and N434Y mutations, according to EU numbering.

[0245] In some embodiments, a variant Fc polypeptide comprises an amino acid sequence having at least 50%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%, or 100% sequence identity to SEQ ID NO: 23, provided that the variant Fc polypeptide comprises a set of mutations corresponding to Y296Q, P329K, L309D, Q311H, and N434S mutations, according to EU numbering.

[0246] In some embodiments, a variant Fc polypeptide comprises an amino acid sequence having at least 50%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%, or 100% sequence identity to SEQ ID NO: 23, provided that the variant Fc polypeptide comprises a set of mutations corresponding to Y296Q, P329K, V309D, Q311H, and N434Y mutations, according to EU numbering.

[0247] In some embodiments, a variant Fc polypeptide comprises an amino acid sequence having at least 50%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%, or 100% sequence identity to SEQ ID NO: 23, provided that the variant Fc polypeptide comprises a set of mutations corresponding to Y296Q, P329K, V309D, Q311H, and N434S mutations, according to EU numbering.

[0248] In some embodiments, a variant Fc polypeptide comprises an amino acid sequence having at least 50%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%, or 100% sequence identity to SEQ ID NO: 23, provided that the variant Fc polypeptide comprises a set of mutations corresponding to L234A, L235A, G237A, Y296Q, P329K, M428L and N434S mutations, according to EU numbering.

[0249] In some embodiments, a variant Fc polypeptide comprises an amino acid sequence having at least 50%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%, or 100% sequence identity to SEQ ID NO: 23, provided that the variant Fc

[0250] -39-

[0251] IPTS / 200163498.2 DOCKET NO. SES-020WO PATENT polypeptide comprises a set of mutations corresponding to L234A, L235A, G237A, Y296Q, P329K, M252Y, S254T, and T256E mutations, according to EU numbering.

[0252] In some embodiments, a variant Fc polypeptide comprises an amino acid sequence having at least 50%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%, or 100% sequence identity to SEQ ID NO: 23, provided that the variant Fc polypeptide comprises a set of mutations corresponding to L234A, L235A, G237A, Y296Q, P329K, L309D, Q311H, and N434Y mutations, according to EU numbering.

[0253] In some embodiments, a variant Fc polypeptide comprises an amino acid sequence having at least 50%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%, or 100% sequence identity to SEQ ID NO: 23, provided that the variant Fc polypeptide comprises a set of mutations corresponding to L234A, L235A, G237A, Y296Q, P329K, L309D, Q311H, and N434S mutations, according to EU numbering.

[0254] In some embodiments, a variant Fc polypeptide comprises an amino acid sequence having at least 50%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%, or 100% sequence identity to SEQ ID NO: 23, provided that the variant Fc polypeptide comprises a set of mutations corresponding to L234A, L235A, G237A, Y296Q, P329K, V309D, Q311H, and N434Y mutations, according to EU numbering.

[0255] In some embodiments, a variant Fc polypeptide comprises an amino acid sequence having at least 50%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%, or 100% sequence identity to SEQ ID NO: 23, provided that the variant Fc polypeptide comprises a set of mutations corresponding to L234A, L235A, G237A, Y296Q, P329K, V309D, Q311H, and N434S mutations, according to EU numbering.

[0256] The mutations and positions of the Fc polypeptide, which can also be referred to as the Fc polypeptide, are according to EU numbering.

[0257] As used herein, a Fc polypeptide / domain comprising a mutation at a specific position is as compared to the wild-type Fc according the numbering system (EU index / EU numbering) as referenced herein.

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[0259] IPTS / 200163498.2 DOCKET NO. SES-020WO PATENT

[0260] In some embodiments, a Fc polypeptide comprises a mutation set of L234A, L235A, G237A, Y296Q, and P329K as compared to SEQ ID NO: 1, wherein the Fc polypeptide further comprises at least, about, or exactly 1-10, 1-20, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15,16, 17, 18, 19 or 20 mutations in addition to the mutation set as compared to SEQ ID NO: 1. Additionally, in some embodiments, the Fc molecule is as provided in the following table.

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[0264] IPTS / 200163498.2 DOCKET NO. SES-020WO PATENT

[0265] In some embodiments, a Fc polypeptide comprises an amino acid sequence having at least 50%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%, or 100% sequence identity to any sequence provided in Table 3. In some embodiments, a Fc polypeptide comprises an amino acid sequence having at least 50%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%, or 100% sequence identity to SEQ ID NO: 11, 12, 13, 19, 20, 21, 22, 23, 24, 25, 42, 43, 44, 45, 46, 47, 48, 49, 200, 201, or 202. In some embodiments, a Fc polypeptide comprises an amino acid sequence having at least 50%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%, or 100% sequence identity to SEQ ID NO: 23. In some embodiments, a Fc polypeptide comprises an amino acid sequence having at least 50%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%, or 100% sequence identity to SEQ ID NO: 24. In some embodiments, a Fc polypeptide comprises an amino acid sequence having at least 50%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%, or 100% sequence identity to SEQ ID NO: 25.

[0266] In some embodiments, a Fc polypeptide comprises an amino acid sequence having at least 50%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%, or 100% sequence identity to any sequence provided in Table 3, provided that the Fc polypeptide comprises one or more mutations at position 234, 235, 237, 265, 269, 296, 298, 329, or any combination thereof. In some embodiments, a Fc polypeptide comprises an amino acid sequence having at least 50%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%, or 100% sequence identity to SEQ ID NO: 19, 20, 21, 22, 23, 24, or 25, provided that the Fc polypeptide comprises one or more mutations at position 234, 235, 237, 265, 269, 296, 298, 329, or any combination thereof. In some embodiments, a Fc polypeptide comprises an amino acid sequence having at least 50%, at least 70%, at least 75%, at least 80%,

[0267] -43-

[0268] IPTS / 200163498.2 DOCKET NO. SES-020WO PATENT at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%, or 100% sequence identity to SEQ ID NO: 23, provided that the Fc polypeptide comprises one or more mutations at position 234, 235, 237, 265, 269, 296, 298, 329, or any combination thereof.

[0269] In some embodiments, a Fc polypeptide comprises an amino acid sequence having at least 50%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%, or 100% sequence identity to any sequence provided in Table 3, and further comprises one or more mutations of L234A, L235A, G237A, Y296Q, P329K, or any combination thereof. In some embodiments, a Fc polypeptide comprises an amino acid sequence having at least 50%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%, or 100% sequence identity to SEQ ID NO: 19, 20, 21, 22, 23, 24, or 25, provided that the Fc polypeptide comprises one or more mutations of L234A, L235A, G237A, Y296Q, P329K, or any combination thereof. In some embodiments, a Fc polypeptide comprises an amino acid sequence having at least 50%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%, or 100% sequence identity to SEQ ID NO: 23, provided that the Fc polypeptide comprises one or more mutations of L234A, L235A, G237A, Y296Q, P329K, or any combination thereof.

[0270] In some embodiments, a Fc polypeptide comprises an amino acid sequence having at least 50%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%, or 100% sequence identity to SEQ ID NO: 19, 20, 21, 22, 23, 24, or 25, provided that the Fc polypeptide comprises a set of mutations corresponding to L234A, L235A, G237A, Y296Q, and P329K mutations. In some embodiments, a Fc polypeptide comprises an amino acid sequence having at least 50%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%, or 100% sequence identity to SEQ ID NO: 23, provided that the Fc polypeptide comprises a set of mutations corresponding to L234A, L235A, G237A, Y296Q, and P329K mutations.

[0271] -44-

[0272] IPTS / 200163498.2 DOCKET NO. SES-020WO PATENT

[0273] In some embodiments, a Fc polypeptide comprises a mutation set of L234A, L235A, G237A, Y296Q, and P329K as compared to SEQ ID NO: 19, 20, 21, 22, 23, 24, or 25, wherein the Fc polypeptide further comprises at least, about, or exactly 1-10, 1-20, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15,16, 17, 18, 19 or 20 mutations in addition to the L234A, L235A, G237A, Y296Q, and P329K mutation set as compared to SEQ ID NO: 19, 20, 21, 22, 23, 24, or 25. In some embodiments, a Fc polypeptide comprises a mutation set of L234A, L235A, G237A, Y296Q, and P329K as compared to SEQ ID NO: 23, wherein the Fc polypeptide further comprises at least, about, or exactly 1-10, 1-20, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15,16, 17, 18, 19 or 20 mutations in addition to the L234A, L235A, G237A, Y296Q, and P329K mutation set as compared to SEQ ID NO: 23.

[0274] In some embodiments, a Fc polypeptide comprises an amino acid sequence as provided in Table 3. In some embodiments, a Fc polypeptide comprises an amino acid sequence of SEQ ID

[0275] NO: 11, 12, 13, 19, 20, 21, 22, 23, 24, 25, 42, 43 , 44, 45, 46, 47, 48, 49, 200, 201, or 202. In some embodiments, a Fc polypeptide comprises an amino acid sequence of SEQ ID NO: 11. In some embodiments, a Fc polypeptide comprises an amino acid sequence of SEQ ID NO: 12. In some embodiments, a Fc polypeptide comprises an amino acid sequence of SEQ ID NO: 13. In some embodiments, a Fc polypeptide comprises an amino acid sequence of SEQ ID NO: 19. In some embodiments, a Fc polypeptide comprises an amino acid sequence of SEQ ID NO: 20. In some embodiments, a Fc polypeptide comprises an amino acid sequence of SEQ ID NO: 21. In some embodiments, a Fc polypeptide comprises an amino acid sequence of SEQ ID NO: 22. In some embodiments, a Fc polypeptide comprises an amino acid sequence of SEQ ID NO: 23. In some embodiments, a Fc polypeptide comprises an amino acid sequence of SEQ ID NO: 24. In some embodiments, a Fc polypeptide comprises an amino acid sequence of SEQ ID NO: 25. In some embodiments, a Fc polypeptide comprises an amino acid sequence of SEQ ID NO: 42. In some embodiments, a Fc polypeptide comprises an amino acid sequence of SEQ ID NO: 43. In some embodiments, a Fc polypeptide comprises an amino acid sequence of SEQ ID NO: 44. In some embodiments, a Fc polypeptide comprises an amino acid sequence of SEQ ID NO: 45. In some embodiments, a Fc polypeptide comprises an amino acid sequence of SEQ ID NO: 46. In some embodiments, a Fc polypeptide comprises an amino acid sequence of SEQ ID NO: 47. In some embodiments, a Fc polypeptide comprises an amino acid sequence of SEQ ID NO: 48. In some embodiments, a Fc polypeptide comprises an amino acid sequence of SEQ ID NO: 49. In

[0276] -45-

[0277] IPTS / 200163498.2 DOCKET NO. SES-020WO PATENT some embodiments, a Fc polypeptide comprises an amino acid sequence of SEQ ID NO: 11 . In some embodiments, a Fc polypeptide comprises an amino acid sequence of SEQ ID NO: 12. In some embodiments, a Fc polypeptide comprises an amino acid sequence of SEQ ID NO: 13. In some embodiments, a Fc polypeptide comprises an amino acid sequence of SEQ ID NO: 200. In some embodiments, a Fc polypeptide comprises an amino acid sequence of SEQ ID NO: 201. In some embodiments, a Fc polypeptide comprises an amino acid sequence of SEQ ID NO: 202.

[0278] In some embodiments, a Fc polypeptide comprises an amino acid sequence having a C- terminal lysine (K). In some embodiments, a Fc polypeptide comprises an amino acid sequence not having a C-terminal lysine (K).

[0279] In some embodiments, the Fc polypeptide, such as those provided herein, is conjugated to another polypeptide. In some embodiments, the another polypeptide is an antibody. In some embodiments, the Fc polypeptide, such as those provided herein, is conjugated to an effector binding / modulating polypeptide or a tissue targeting polypeptide. In some embodiments, the Fc polypeptide, such as those provided herein, is conjugated to an antibody, or antigen-binding fragment thereof. In some embodiments, the Fc polypeptide, such as those, is conjugated to another polypeptide via a linker or a covalent bond. In some embodiments, the linker is a protein linker, such as those provided herein.

[0280] In some embodiments, the proteases and polypeptides provided for herein are linked to a half-life extension moiety.

[0281] In some embodiments, the half-life extension moiety is albumin, such as human serum albumin (HSA). In some embodiments, the albumin is as provided for in US20200392206A1, US20180200346A1, US20180265569A1, US20180265570A1, US20200102367A1, US8822417B2, US10633428B2. The albumin can be linked to the protease molecule, either directly or via a peptide linker, such as those, but not limited to, provided for herein. The albumin molecule could also be used in conjunction with a Fc molecule. However, in some embodiments, the half-life extension moiety does not comprise a Fc molecule, which can also be referred to as a Fc polypeptide domain.

[0282] In some embodiments, the half-life extension moiety is an antibody, such as a nanobody or single variable domain, which binds to HSA. Examples of such nanobodies include, but are not limited to, those that are provided for in PCT Publication No. WO2017085172, U.S. Publication No. 20220380445A1, U.S. Patent No. 11,414,480, PCT Publication No.

[0283] -46-

[0284] IPTS / 200163498.2 DOCKET NO. SES-020WO PATENT

[0285] WO2018104444A1, U.S. Publication No. 20230060574A1, PCT Publication No. WO2018134235A1, U.S. Patent No. 11,897,944, including Alb-8, Alb-23, or variants thereof, provided for therein, each of which is hereby incorporated by reference in its entirety. The HSA antibody can be linked to the protease in various formats. In some embodiments the antibody is linked to the protease through a Fc domain, such as those provided for herein. In some embodiments, the Fc is protease resistant, which are also described in U.S. Application No. 18 / 512,556, which is incorporated by reference in its entirety.

[0286] Proteases and Variants thereof

[0287] The present disclosure also provides for polypeptides, molecules, compounds, therapeutics, and compositions comprising a polypeptide having protease activity, which can be covalently or non-covalently connected to a Fc polypeptide domain or other domain (e g. halflife extension moiety), such as those provided for herein. In some embodiments, the polypeptide having protease activity is IdeS, IdeSsuis, IdeZ, IdeE, IdeE2, IdeZ2, or IdeC. The protease can be referred to as an immunoglobulin protease.

[0288] Streptococcus pyogenes is a significant bacterial pathogen that secretes two enzymes showing remarkable specificity for IgG; EndoS and IdeS. EndoS (Endoglycosidase in Streptococcus pyogenes) specifically hydrolyzes the functionally important N-linked glycan of IgG, and treatment with EndoS abrogates the pathogenic activity of IgG in mouse models of autoimmune disease. (Collin M, Olsen A. EndoS, a novel secreted protein from Streptococcus pyogenes with endoglycosidase activity on human IgG. Embo J. 2001;20:3046-3055; Nandakumar KS, Collin M, Olsen A, Nimmerjahn F, Blom AM, et al. Endoglycosidase treatment abrogates IgG arthritogenicity: importance of IgG glycosylation in arthritis. Eur J Immunol. 2007;37:2973-2982) IdeS (Immunoglobulin G-degrading enzyme of Streptococcus pyogenes) is a cysteine proteinase which cleaves IgG with a unique degree of specificity in the hinge region. (Wenig K, Chatwell L, von Pawel-Rammingen U, Bjbrck L, Huber R, et al. Structure of the streptococcal endopeptidase IdeS, a cysteine proteinase with strict specificity for IgG. Proc Natl Acad Sci USA. 2004; 101 : 17371-17376) IdeS is extremely specific for IgG, which is hydrolyzed in the hinge region after glycine residue 236 in both heavy chains, which generates one F(ab')2 and two monomeric Fc fragments. IdeE is a homolog of the secreted IgG-

[0289] -47-

[0290] IPTS / 200163498.2 DOCKET NO. SES-020WO PATENT specific protease IdeS / Mac of Streptococcus pyogenes. The activity of IdeE is comparable with the activity of IdeZ, the corresponding enzyme of the closely related S. equi ssp. zooepidemicus .

[0291] The full sequence of IdeS is publicly available as NCBI Reference Sequence no.

[0292] WP_010922160.1 and is provided herein as SEQ ID NO: 35:

[0293] MRKRCYSTSAAVLAAVTLFVLSVDRGVIADSFSANQEIRYSEVTPYHVTSVWTKGVTPP ANFTQGEDVFHAPYVANQGWYDITKTFNGKDDLLCGAATAGNMLHWWFDQNKDQIKRYL EEHPEKQKINFNGEQMFDVKEAIDTKNHQLDSKLFEYFKEKAFPYLSTKHLGVFPDHVI DMFINGYRLSLTNHGPTPVKEGSKDPRGGI FDAVFTRGDQSKLLTSRHDFKEKNLKEIS DLIKKELTEGKALGLSHTYANVRINHVINLWGADFDSNGNLKAIYVTDSDSNASIGMKK YFVGVNSAGKVAISAKEIKEDNIGAQVLGLFTLSTGQDSWNQTN ( SEQ ID NO : 35 ) .

[0294] This sequence includes an N-terminus methionine followed by a 28 amino acid secretion signal sequence. The N-terminus methionine and the signal sequence (a total of 29 amino acids at the N terminus) are typically removed to form the mature IdeS protein, the amino acid sequence of which is publicly available as UniProt Identifier Q9F1R7 STRPY and is provided herein as SEQ ID NO: 36. Another variant of an IdeS protease that can be used as a reference sequence can be as illustrated in SEQ ID NO: 171, which is similar to the amino acid sequence of SEQ ID NO: 36, but has an additional 4 amino acid residues at the N-terminus, which are DSFS (SEQ ID NO: 220).

[0295] IdeZ is an IgG cysteine protease produced by Streptococcus equi ssp. zooepidemicus, a bacterium predominantly found in horses. As IdeZ is not a human pathogen, human subjects do not typically have antibodies against this protein in their plasma. However, IdeZ has a level of IgG cysteine protease activity against human IgG which is considerably lower than that of IdeS. The full sequence of IdeZ is publicly available as UniProt Identifier Q0PIW1 and is provided herein as SEQ ID NO: 107 below:

[0296] MKTIAYPNKPHSLSAGLLTAIAI FSLASSNITYADDYQRNAAEVYAKEVPHQIT SVWTKGVTPLTPEQFRYNNEDVIHAPYLAHQGWYDITKVFDGKDNLLCGAATAG NMLHWWFDQNKTEIEAYLSKHPEKQKI IFNNQELFDLKAAIDTKDSQTNSQLFN YFRDKAFPNLSARQLGVMPDLVLDMFINGYYLNVFKTQSTDVNRPYQDKDKRGG I FDAVFTRGDQTTLLTARHDLKNKGLNDI S T I IKQELTEGRALALSHT YANVS I S HVINLWGAD FNAE GNLEAI YVT DS DANAS I GMKKY FVG I NAHGHVAI S AKK I E GENIGAQVLGLFTLSSGKDIWQKLS (SEQ ID NO: 107).

[0297] In some embodiments, IdeZ is publicly available as NCBI Reference Sequence no WP_014622780.1 and is provided herein as SEQ ID NO: 108 below:

[0298] -48-

[0299] IPTS / 200163498.2 DOCKET NO. SES-020WO PATENT

[0300] MKTIAYPNKPHSLSAGLLTAIAI FSLASSNITYADDYQRNATEAYAKEVPHQIT SVWTKGVIPLIPEQFRYNNEDVIHAPYLAHQGWYDITKAFDGKDNLLCGAATAG NMLHWWFDQNKTEIEAYLSKHPEKQKI IFNNQELFDLKAAIDTKDSQINSQLFN YFRDKAFPNLSARQLGVMPDLVLDMFINGYYLNVFKIQSTDVNRPYQDKDKRGG I FDAVFIRGDQTILLTARHDLKNKGLNDISTI IKQELTEGRALALSHIYANVS I S HVINLWGAD FNAE GNLEAI YVI DS DANAS I GMKKY FVG I NAHGHVAI S AKK I E GENIGAQVLGLFTLSSGKDIWQKLS (SEQ ID NO: 108).

[0301] The mature sequence of IdeZ is publicly available as UniProt Identifier A0A0D0YKS5 STRSZ and is provided herein as SEQ ID NO: 38.

[0302] In some embodiments, the polypeptide having protease activity is IdeS, IdeSsuis, IdeZ, IdeE, IdeE2, IdeZ2, Ide85, or IdeC. In some embodiments, the IdeS, IdeZ, IdeE, IdeE2, IdeZ2, Ide85, or IdeC protease has an amino acid sequence such as those provided herein. The mature sequence of IdeSsuis is publicly available as UniProt Identifier C5W022 and is provided herein as SEQ ID NO: 55. The mature sequence of IdeE is publicly available as UniProt Identifier C0M8U6 STRE4 and is provided herein as SEQ ID NO: 37. The mature sequence of IdeE2 is publicly available as UniProt Identifier C7B615 9STRE and is provided herein as SEQ ID NO: 39. The mature sequence of IdeZ2 is publicly available as UniProt Identifier B4U2F7 STREM and is provided herein as SEQ ID NO: 40. The mature sequence of IdeC is publicly available as UniProt Identifier A0A3P5YAY8 STRCB and is provided herein as SEQ ID NO: 41. In some embodiments, the protease is an IgG degrading protease. In some embodiments, the protease is an IgM degrading protease.

[0303] -49-

[0304] IPTS / 200163498.2 DOCKET NO. SES-020WO PATENT

[0305] -50-

[0306] IPTS / 200163498.2 DOCKET NO. SES-020WO PATENT

[0307] In some embodiments, the protease provided for herein does not comprises the amino acid sequence of the polypeptides in Table 4.

[0308] In some embodiments, a polypeptide having protease activity comprises an amino acid sequence having at least 50%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%, or 100% sequence identity to any sequence provided in Table 4. In some embodiments, the polypeptide having protease activity is IdeS. In some embodiments, the polypeptide having protease activity is IdeSsuis. In some embodiments, the polypeptide having protease activity is IdeE. In some embodiments, the polypeptide having protease activity is IdeZ.

[0309] -51-

[0310] IPTS / 200163498.2 DOCKET NO. SES-020WO PATENT

[0311] In some embodiments, the polypeptide having protease activity is IdeE2. In some embodiments, the polypeptide having protease activity is IdeZ2. In some embodiments, the polypeptide having protease activity is Ide85. In some embodiments, the polypeptide having protease activity is IdeC. In some embodiments, a polypeptide having protease activity comprises an amino acid sequence having at least 50%, 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%, or 100% sequence identity to SEQ ID NO: 26, 28, 29, 31, 36, 37, 38, 39, 40, 41, 50, 51, 52, or 55.

[0312] In some embodiments, a polypeptide having protease activity comprises an amino acid sequence as provided in Table 4. In some embodiments, the polypeptide having protease activity is IdeS. In some embodiments, the polypeptide having protease activity is IdeSsuis. In some embodiments, the polypeptide having protease activity is IdeE. In some embodiments, the polypeptide having protease activity is IdeZ. In some embodiments, the polypeptide having protease activity is IdeE2. In some embodiments, the polypeptide having protease activity is IdeZ2. In some embodiments, the polypeptide having protease activity is Ide85. In some embodiments, the polypeptide having protease activity is IdeC. In some embodiments, a polypeptide having protease activity comprises an amino acid sequence of SEQ ID NO: 26, 28, 29, 31, 36, 37, 38, 39, 40, 41, 50, 51, 52, or 55.

[0313] In some embodiments, the protease comprises an amino acid sequence that comprises a signal sequence. In some embodiments, the protease comprises an amino acid sequence that does not comprise a signal sequence. In some embodiments, the signal sequence may have the amino acid sequence of METDTLLLWVLLLWVPGSTG (SEQ ID NO: 109). Thus, in some embodiments, the mature protein does not comprise a signal sequence, such as the amino acid sequence of SEQ ID NO: 109.

[0314] In some embodiments, the polypeptide or protease has a sequence selected from Table 5.

[0315] -52-

[0316] IPTS / 200163498.2 DOCKET NO. SES-020WO PATENT

[0317] -53-

[0318] IPTS / 200163498.2 DOCKET NO. SES-020WO PATENT

[0319] In some embodiments, the protease, such as an IgG protease, comprises a mutation at position 163 as compared to SEQ ID NO: 35. In some embodiments, the protease comprises a Y163D mutation, which can be referred to as an aspartic acid (D) at position 163 as compared to SEQ ID NO: 35. In some embodiments, the mutation is a Y163E mutation. In some embodiments, a pharmaceutical composition is provided comprising an IgG protease comprising the Y163D mutation, wherein the IgG protease does not aggregate or produce high molecular weight complexes as compared to an IgG protease that does not have a mutation at position 163, such as, but not limited to Y163D. In some embodiments, the mutation can be Y163N. In some embodiments, the pharmaceutical composition has less than 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, or 1% high molecular weight complexes, which can represent aggregates. Accordingly, in some embodiments, an IgG protease is provided that has a set of mutations as provided for herein, and optionally a Y163D mutation. In some embodiments, the protease comprises a sequence that is at least 50%, at least 60%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identical to SEQ ID NO: 35 and comprises a mutation at position 163, including but not limited to Y163D or Y163N. In some embodiments, the mutation can be Y163E. Although certain protein variants provided for herein may comprise a different mutation or no mutation at all at position 163, the embodiments provided for herein also provide for such variants comprising a Y163D, Y163N, or Y163E mutation. In some embodiments, a protease variant is provided with reduced aggregation as compared to its reference sequence. The reference sequence can be any Ig protease provided for herein or known to one of skill in the art. Accordingly, in some embodiments, the 1g protease (e.g., IgG protease) can comprise a mutation that corresponds to a mutation at position 163 of SEQ ID NO: 35. In

[0320] -54-

[0321] IPTS / 200163498.2 DOCKET NO. SES-020WO PATENT some embodiments, the mutation is a Y163D mutation. In some embodiments, the mutation is a Y163E mutation. In some embodiments, the mutation is a Y163N mutation. The reference sequence can also be as provided for in U.S. Patent No. 12,359,183, U.S. Patent No. 12,129,499, and PCT Publication No. WO2024206953, each of which is hereby incorporated by reference in its entirety. In some embodiments, the protease is at least 50%, at least 60%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identical to the proteases provided for in U.S. Patent No. 12,359,183, U.S. Patent No. 12,129,499, and PCT Publication No. WO2024206953, each of which is hereby incorporated by reference in its entirety.

[0322] In some embodiments, the polypeptide or protease has an amino acid sequence having at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to any one amino acid sequence selected from SEQ ID NO: 57, 58, 59, 60, 189, 190, 191, 192, 193, 194, 195, 196, 197, 198, or 199. Additional support for specific embodiments of polypeptides having an amino acid sequence having at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to any one amino acid sequence selected from SEQ ID NO: 57, 58, 59, 60, 189, 190, 191, 192, 193, 194, 195, 196, 197, 198, or 199 may be found in U.S. Provisional Application No. 63 / 701,666, filed October 1, 2024, U.S. Provisional Application No. 63 / 478,789, filed January 6, 2023, U.S. Provisional Application No. 63 / 483,142, filed February 3, 2023, U.S. Provisional Application No. 63 / 493,142, filed March 31, 2023, U.S. Provisional Application No. 63 / 506,539, filed June 6, 2023, and / or U.S. Provisional Application No. 63 / 600,157, filed November 17, 2023, each of which is hereby incorporated by reference in its entirety.

[0323] In some embodiments, the polypeptide has an amino acid sequence having at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to the amino acid sequence of SEQ ID NO: 57. In some embodiments, the polypeptide has an amino acid sequence having at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least

[0324] -55-

[0325] IPTS / 200163498.2 DOCKET NO. SES-020WO PATENT

[0326] 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to the amino acid sequence of SEQ ID NO: 58. In some embodiments, the polypeptide has an amino acid sequence having at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to the amino acid sequence of SEQ ID NO: 59. In some embodiments, the polypeptide has an amino acid sequence having at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to the amino acid sequence of SEQ ID NO: 60. In some embodiments, the polypeptide has an amino acid sequence having at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to the amino acid sequence of SEQ ID NO: 189. In some embodiments, the polypeptide has an amino acid sequence having at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to the amino acid sequence of SEQ ID NO: 190. In some embodiments, the polypeptide has an amino acid sequence having at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to the amino acid sequence of SEQ ID NO: 191. In some embodiments, the polypeptide has an amino acid sequence having at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to the amino acid sequence of SEQ ID NO: 192. In some embodiments, the polypeptide has an amino acid sequence having at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to the amino acid sequence of SEQ ID NO: 193. In some embodiments, the polypeptide has an amino acid sequence having at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to the amino acid sequence of SEQ ID NO: 194. In some embodiments, the polypeptide

[0327] -56-

[0328] IPTS / 200163498.2 DOCKET NO. SES-020WO PATENT has an amino acid sequence having at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to the amino acid sequence of SEQ ID NO: 195. In some embodiments, the polypeptide has an amino acid sequence having at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to the amino acid sequence of SEQ ID NO: 196. In some embodiments, the polypeptide has an amino acid sequence having at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to the amino acid sequence of SEQ ID NO: 197. In some embodiments, the polypeptide has an amino acid sequence having at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to the amino acid sequence of SEQ ID NO: 198. In some embodiments, the polypeptide has an amino acid sequence having at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to the amino acid sequence of SEQ ID NO: 199.

[0329] In some embodiments, the polypeptide has an amino acid sequence having at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to the amino acid sequence of SEQ ID NO: 57, provided that the polypeptide comprises a threonine (T) at position 60, an aspartic acid (D) at position 61, an isoleucine (I) at position 63, a tyrosine (Y) at position 64, a leucine (L) at position 69, a lysine (K) at position 74, a glycine (G) at position 76, a leucine (L) at position 85, a glutamic acid (E) at position 112, an asparagine (N) at position 116, a lysine (K) at position 119, a lysine (K) at position 120, a tyrosine (Y) at position 121, a glycine (G) at position 130, a leucine (L) at position 134, an aspartic acid (D) at position 145, a serine (S) at position 146, a threonine (T) at position 148, an aspartic acid (D) at position 154, an aspartic acid (D) at position 190, an arginine (R) at position 198, an asparagine (N) at position 223, a tyrosine (Y) at position 225, a leucine (L) at position 227, a threonine (T) at position 231, a lysine (K) at position 241, an isoleucine (I) at position

[0330] -57-

[0331] IPTS / 200163498.2 DOCKET NO. SES-020WO PATENT

[0332] 251 , a glycine (G) at position 261, an alanine (A) at position 273, a glutamic acid (E) at position 274, a glutamic acid (E) at position 278, a proline (P) at position 289, a leucine (L) at position 303, a glutamic acid (E) at position 314, and an isoleucine (I) at position 334, wherein the positions correspond to the numbering as set forth in SEQ ID NO: 35.

[0333] In some embodiments, the polypeptide has an amino acid sequence having at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to the amino acid sequence of SEQ ID NO: 58, provided that the polypeptide comprises a threonine (T) at position 60, an aspartic acid (D) at position 61, an isoleucine (I) at position 63, a tyrosine (Y) at position 64, a leucine (L) at position 69, a lysine (K) at position 74, a glycine (G) at position 76, a leucine (L) at position 85, a glutamic acid (E) at position 112, an asparagine (N) at position 116, a lysine (K) at position 119, a lysine (K) at position 120, a tyrosine (Y) at position 121, a glycine (G) at position 130, a leucine (L) at position 134, an aspartic acid (D) at position 145, a serine (S) at position 146, a threonine (T) at position 148, an aspartic acid (D) at position 154, an aspartic acid (D) at position 163, an aspartic acid (D) at position 190, an arginine (R) at position 198, an asparagine (N) at position 223, a tyrosine (Y) at position 225, a leucine (L) at position 227, a threonine (T) at position 231, a lysine (K) at position 241, an isoleucine (I) at position 251, a glycine (G) at position 261, an alanine (A) at position 273, a glutamic acid (E) at position 274, a glutamic acid (E) at position 278, a proline (P) at position 289, a leucine (L) at position 303, a glutamic acid (E) at position 314, and an isoleucine (I) at position 334, wherein the positions correspond to the numbering as set forth in SEQ ID NO: 35.

[0334] In some embodiments, the polypeptide has an amino acid sequence having at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to the amino acid sequence of SEQ ID NO: 59, provided that the polypeptide comprises a threonine (T) at position 60, an aspartic acid (D) at position 61, an isoleucine (I) at position 63, a tyrosine (Y) at position 64, a leucine (L) at position 69, a lysine (K) at position 74, a glycine (G) at position 76, a leucine (L) at position 85, a glutamic acid (E) at position 112, an asparagine (N) at position 116, a lysine (K) at position 119, a lysine (K) at position 120, a tyrosine (Y) at position 121, a glycine (G) at position 130, a leucine (L) at position 134, an

[0335] -58-

[0336] IPTS / 200163498.2 DOCKET NO. SES-020WO PATENT aspartic acid (D) at position 145, a serine (S) at position 146, a threonine (T) at position 148, an aspartic acid (D) at position 154, an asparagine (N) at position 163, an alanine (A) at position 166, an arginine (R) at position 167, a glutamine (Q) at position 168, an aspartic acid (D) at position 190, an arginine (R) at position 198, an asparagine (N) at position 223, a tyrosine (Y) at position 225, a leucine (L) at position 227, a threonine (T) at position 231, a lysine (K) at position 241, an isoleucine (I) at position 251, a glycine (G) at position 261, an alanine (A) at position 273, a glutamic acid (E) at position 274, a glutamic acid (E) at position 278, a proline (P) at position 289, a leucine (L) at position 303, a glutamic acid (E) at position 314, and an isoleucine (I) at position 334, wherein the positions correspond to the numbering as set forth in SEQ ID NO: 35.

[0337] In some embodiments, the polypeptide has an amino acid sequence having at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to the amino acid sequence of SEQ ID NO: 60, provided that the polypeptide comprises a threonine (T) at position 60, an aspartic acid (D) at position 61, an isoleucine (I) at position 63, a tyrosine (Y) at position 64, a leucine (L) at position 69, a lysine (K) at position 74, a glycine (G) at position 76, a leucine (L) at position 85, a glutamic acid (E) at position 112, an asparagine (N) at position 116, a lysine (K) at position 119, a lysine (K) at position 120, a tyrosine (Y) at position 121, a glycine (G) at position 130, a leucine (L) at position 134, an aspartic acid (D) at position 145, a serine (S) at position 146, a threonine (T) at position 148, an aspartic acid (D) at position 154, an asparagine (N) at position 163, an alanine (A) at position 166, an arginine (R) at position 167, a glutamine (Q) at position 168, an isoleucine (I) at position 169, an aspartic acid (D) at position 190, an arginine (R) at position 198, an asparagine (N) at position 223, a tyrosine (Y) at position 225, a leucine (L) at position 227, a threonine (T) at position 231, a lysine (K) at position 241, an isoleucine (I) at position 251, a glycine (G) at position 261, an alanine (A) at position 273, a glutamic acid (E) at position 274, a glutamic acid (E) at position 278, a proline (P) at position 289, a leucine (L) at position 303, a glutamic acid (E) at position 314, and an isoleucine (I) at position 334, wherein the positions correspond to the numbering as set forth in SEQ ID NO: 35.

[0338] In some embodiments, the polypeptide has an amino acid sequence selected from SEQ ID NO: 57, 58, 59, 60, 189, 190, 191, 192, 193, 194, 195, 196, 197, 198, or 199. In some

[0339] -59-

[0340] IPTS / 200163498.2 DOCKET NO. SES-020WO PATENT embodiments, the polypeptide has an amino acid sequence of SEQ ID NO: 57. Tn some embodiments, the polypeptide has an amino acid sequence of SEQ ID NO: 58. In some embodiments, the polypeptide has an amino acid sequence of SEQ ID NO: 59. In some embodiments, the polypeptide has an amino acid sequence of SEQ ID NO: 60. In some embodiments, the polypeptide has an amino acid sequence of SEQ ID NO: 189. In some embodiments, the polypeptide has an amino acid sequence of SEQ ID NO: 190. In some embodiments, the polypeptide has an amino acid sequence of SEQ ID NO: 191. In some embodiments, the polypeptide has an amino acid sequence of SEQ ID NO: 192. In some embodiments, the polypeptide has an amino acid sequence of SEQ ID NO: 193. In some embodiments, the polypeptide has an amino acid sequence of SEQ ID NO: 194. In some embodiments, the polypeptide has an amino acid sequence of SEQ ID NO: 195. In some embodiments, the polypeptide has an amino acid sequence of SEQ ID NO: 196. In some embodiments, the polypeptide has an amino acid sequence of SEQ ID NO: 197. In some embodiments, the polypeptide has an amino acid sequence of SEQ ID NO: 198. In some embodiments, the polypeptide has an amino acid sequence of SEQ ID NO: 199.

[0341] In some embodiments, the protease and polypeptides provided for herein comprise a leader sequence that is present at the N-terminus of the polypeptide. Without being bound to any particular theory, leader sequences can facilitate the expression of polypeptides in eukaryotic cells. Any suitable leader sequence can be utilized. For example, leader sequences comprising the sequence of METDTLLLWVLLLWVPGSTG (SEQ ID NO: 109) may be used. These are non-limiting examples of leader sequences. The leader sequences are processed (removed) from the polypeptide as it is being translated and expressed in the cell leaving the polypeptides provided for herein. Thus, in some embodiments, when present in a pharmaceutical composition or as an isolated polypeptide, the polypeptides provided for herein do not comprise a leader sequence. In some embodiments, polypeptides provided for herein comprise an amino acid sequence having at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to any one amino acid sequence selected from SEQ ID NO: 57, 58, 59, 60, 189, 190, 191, 192, 193, 194, 195, 196, 197, 198, or 199, and further comprises a leader amino acid sequence. In some embodiments, the protease has an amino acid sequence having at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%,

[0342] -60-

[0343] IPTS / 200163498.2 DOCKET NO. SES-020WO PATENT at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to any one amino acid sequence selected from SEQ ID NO: 57, 58, 59, 60, 189, 190, 191, 192, 193, 194, 195, 196, 197, 198, or 199, and further comprises a leader amino acid sequence having an amino acid sequence of METDTLLLWVLLLWVPGSTG (SEQ ID NO: 109). In some embodiments, the leader amino acid sequence is N-terminal to the protease. In some embodiments, the leader amino acid sequence facilitates expression of the protease. In some embodiments, the leader amino acid sequence is processed and removed from the protease during expression ex vivo or in vivo.

[0344] In some embodiments, the protease comprises the polypeptide. In some embodiments, the polypeptide comprises a mutation at any position as compared to SEQ ID NO: 35. In some embodiments, the mutation is a substitution, a deletion, or an insertion. In some embodiments, the mutation is a substitution.

[0345] In some embodiments, the polypeptide comprises the amino acid sequence having at least 50%, at least 60%, at least 70%, at least 80%, at least 81%, at least 82%, at least 83%, at least

[0346] 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least

[0347] 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least

[0348] 98%, or at least 99% sequence identity to SEQ ID NO: 35, and having a mutation at a position selected from 60, 61, 63, 64, 69, 74, 76, 85, 112, 116, 119, 120, 121, 130, 134, 145, 146, 148, 154, 190, 198, 223, 225, 227, 231, 251, 261, 273, 274, 278, 289, 303, 314, 334, but not position 241, or any combination thereof, as compared to SEQ ID NO: 35.

[0349] In some embodiments, the polypeptide comprises the amino acid sequence having at least 50%, at least 60%, at least 70%, at least 80%, at least 81%, at least 82%, at least 83%, at least

[0350] 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least

[0351] 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least

[0352] 98%, or at least 99% sequence identity to SEQ ID NO: 35, and having a set of mutations at positions 60, 61, 63, 64, 69, 74, 76, 85, 112, 116, 119, 120, 121, 130, 134, 145, 146, 148, 154, 163, 166, 167, 168, 169, 190, 198, 223, 225, 227, 231, 251, 261, 273, 274, 278, 289, 303, 314, and 334, but not position 241, as compared to SEQ ID NO: 35.

[0353] In some embodiments, the polypeptide comprises the amino acid sequence having at least 50%, at least 60%, at least 70%, at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least

[0354] -61-

[0355] IPTS / 200163498.2 DOCKET NO. SES-020WO PATENT

[0356] 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 35, and having a set of mutations at positions 60, 61, 63, 64, 69, 74, 76, 85, 112, 116, 119, 120, 121, 130, 134, 145, 146, 148, 154, 163, 190, 198, 223, 225, 227, 231, 251, 261, 273, 274, 278, 289, 303, 314, and 334, but not position 241, as compared to SEQ ID NO: 35.

[0357] In some embodiments, the polypeptide comprises the amino acid sequence having at least 50%, at least 60%, at least 70%, at least 80%, at least 81%, at least 82%, at least 83%, at least

[0358] 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least

[0359] 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least

[0360] 98%, or at least 99% sequence identity to SEQ ID NO: 35, and having a set of mutations at positions 60, 61, 63, 64, 69, 74, 76, 85, 112, 116, 119, 120, 121, 130, 134, 145, 146, 148, 154, 163, 166, 167, 168, 190, 198, 223, 225, 227, 231, 251, 261, 273, 274, 278, 289, 303, 314, and 334, but not position 241, as compared to SEQ ID NO: 35.

[0361] In some embodiments, the polypeptide comprises the amino acid sequence having at least 50%, at least 60%, at least 70%, at least 80%, at least 81%, at least 82%, at least 83%, at least

[0362] 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least

[0363] 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least

[0364] 98%, or at least 99% sequence identity to SEQ ID NO: 35, and having a set of mutations at positions 60, 61, 63, 64, 69, 74, 76, 85, 112, 116, 119, 120, 121, 130, 134, 145, 146, 148, 154, 163, 166, 167, 168, 169, 190, 198, 223, 225, 227, 231, 251, 261, 273, 274, 278, 289, 303, 314, and 334, but not position 241, as compared to SEQ ID NO: 35.

[0365] In some embodiments, the polypeptide has an amino acid sequence having at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to the amino acid sequence of SEQ ID NO: 35, provided that the polypeptide comprises a threonine (T) at position 60, an aspartic acid (D) at position 61, an isoleucine (I) at position 63, a tyrosine (Y) at position 64, a leucine (L) at position 69, a lysine (K) at position 74, a glycine (G) at position 76, a leucine (L) at position 85, a glutamic acid (E) at position 112, an asparagine (N) at position 116, a lysine (K) at position 119, a lysine (K) at position 120, a tyrosine (Y) at position 121, a glycine (G) at position 130, a leucine (L) at position 134, an aspartic acid (D) at position 145, a serine (S) at position 146, a threonine (T) at position 148, an

[0366] -62-

[0367] IPTS / 200163498.2 DOCKET NO. SES-020WO PATENT aspartic acid (D) at position 154, an aspartic acid (D) at position 190, an arginine (R) at position 198, an asparagine (N) at position 223, a tyrosine (Y) at position 225, a leucine (L) at position 227, a threonine (T) at position 231, a lysine (K) at position 241, an isoleucine (I) at position 251, a glycine (G) at position 261, an alanine (A) at position 273, a glutamic acid (E) at position 274, a glutamic acid (E) at position 278, a proline (P) at position 289, a leucine (L) at position 303, a glutamic acid (E) at position 314, and an isoleucine (I) at position 334, wherein the positions correspond to the numbering as set forth in SEQ ID NO: 35.

[0368] In some embodiments, the polypeptide has an amino acid sequence having at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to the amino acid sequence of SEQ ID NO: 35, provided that the polypeptide comprises a threonine (T) at position 60, an aspartic acid (D) at position 61, an isoleucine (I) at position 63, a tyrosine (Y) at position 64, a leucine (L) at position 69, a lysine (K) at position 74, a glycine (G) at position 76, a leucine (L) at position 85, a glutamic acid (E) at position 112, an asparagine (N) at position 116, a lysine (K) at position 119, a lysine (K) at position 120, a tyrosine (Y) at position 121, a glycine (G) at position 130, a leucine (L) at position 134, an aspartic acid (D) at position 145, a serine (S) at position 146, a threonine (T) at position 148, an aspartic acid (D) at position 154, an aspartic acid (D) at position 163, an aspartic acid (D) at position 190, an arginine (R) at position 198, an asparagine (N) at position 223, a tyrosine (Y) at position 225, a leucine (L) at position 227, a threonine (T) at position 231, a lysine (K) at position 241, an isoleucine (I) at position 251, a glycine (G) at position 261, an alanine (A) at position 273, a glutamic acid (E) at position 274, a glutamic acid (E) at position 278, a proline (P) at position 289, a leucine (L) at position 303, a glutamic acid (E) at position 314, and an isoleucine (I) at position 334, wherein the positions correspond to the numbering as set forth in SEQ ID NO: 35.

[0369] In some embodiments, the polypeptide has an amino acid sequence having at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to the amino acid sequence of SEQ ID NO: 35, provided that the polypeptide comprises a threonine (T) at position 60, an aspartic acid (D) at position 61, an isoleucine (I) at position 63, a tyrosine (Y) at position 64, a leucine (L) at position 69, a lysine (K) at position 74,

[0370] -63-

[0371] IPTS / 200163498.2 DOCKET NO. SES-020WO PATENT a glycine (G) at position 76, a leucine (L) at position 85, a glutamic acid (E) at position 112, an asparagine (N) at position 116, a lysine (K) at position 119, a lysine (K) at position 120, a tyrosine (Y) at position 121, a glycine (G) at position 130, a leucine (L) at position 134, an aspartic acid (D) at position 145, a serine (S) at position 146, a threonine (T) at position 148, an aspartic acid (D) at position 154, an asparagine (N) at position 163, an alanine (A) at position 166, an arginine (R) at position 167, a glutamine (Q) at position 168, an aspartic acid (D) at position 190, an arginine (R) at position 198, an asparagine (N) at position 223, a tyrosine (Y) at position 225, a leucine (L) at position 227, a threonine (T) at position 231, a lysine (K) at position 241, an isoleucine (I) at position 251, a glycine (G) at position 261, an alanine (A) at position 273, a glutamic acid (E) at position 274, a glutamic acid (E) at position 278, a proline (P) at position 289, a leucine (L) at position 303, a glutamic acid (E) at position 314, and an isoleucine (I) at position 334, wherein the positions correspond to the numbering as set forth in SEQ ID NO: 35.

[0372] In some embodiments, the polypeptide has an amino acid sequence having at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to the amino acid sequence of SEQ ID NO: 35, provided that the polypeptide comprises a threonine (T) at position 60, an aspartic acid (D) at position 61, an isoleucine (I) at position 63, a tyrosine (Y) at position 64, a leucine (L) at position 69, a lysine (K) at position 74, a glycine (G) at position 76, a leucine (L) at position 85, a glutamic acid (E) at position 112, an asparagine (N) at position 116, a lysine (K) at position 119, a lysine (K) at position 120, a tyrosine (Y) at position 121, a glycine (G) at position 130, a leucine (L) at position 134, an aspartic acid (D) at position 145, a serine (S) at position 146, a threonine (T) at position 148, an aspartic acid (D) at position 154, an asparagine (N) at position 163, an alanine (A) at position 166, an arginine (R) at position 167, a glutamine (Q) at position 168, an isoleucine (I) at position 169, an aspartic acid (D) at position 190, an arginine (R) at position 198, an asparagine (N) at position 223, a tyrosine (Y) at position 225, a leucine (L) at position 227, a threonine (T) at position 231, a lysine (K) at position 241, an isoleucine (I) at position 251, a glycine (G) at position 261, an alanine (A) at position 273, a glutamic acid (E) at position 274, a glutamic acid (E) at position 278, a proline (P) at position 289, a leucine (L) at position 303, a glutamic acid

[0373] -64-

[0374] IPTS / 200163498.2 DOCKET NO. SES-020WO PATENT

[0375] (E) at position 314, and an isoleucine (I) at position 334, wherein the positions correspond to the numbering as set forth in SEQ ID NO: 35.

[0376] In some embodiments, the polypeptide comprises a lysine (K) at position 241, as compared to SEQ ID NO: 35.

[0377] In some embodiments, the polypeptide does not comprise the amino acid sequence of DSFSANQEIRYSEVTPYHVT (SEQ ID NO: 110). In some embodiments, the polypeptide does not include the amino acid sequence of DSFSANQEIRYSEVTPYHVT (SEQ ID NO: 110). In some embodiments, the polypeptide does not include the contiguous amino acid sequence of DSFSANQEIRYSEVTPYHVT (SEQ ID NO: 110) in its entirety. In some embodiments, the polypeptide does not include the contiguous amino acid sequence of DSFSANQEIRYSEVTPYHVT (SEQ ID NO: 110) in its entirety, or the dipeptide of VT at the N-terminus of the polypeptide. As used herein, the N-terminus of the polypeptide in reference to the contiguous amino acid sequence of DSFSANQEIRYSEVTPYHVT (SEQ ID NO: 110), or the dipeptide of VT means the N-terminus of the protease, which may be C-terminal to a linker or another polypeptide if the protease is linked to another polypeptide, such as the Fc domains provided for herein. In some embodiments, the polypeptide does not include the dipeptide of VT at the N-terminus of the polypeptide. In some embodiments, the polypeptide does not comprise the amino acid sequence of DSFSANQEIRYSEVTPYH (SEQ ID NO: 111). In some embodiments, the polypeptide does not comprise the amino acid sequence of ANQEIRYSEVTPYHVT (SEQ ID NO: 112). In some embodiments, the polypeptide does not comprise the amino acid sequence of ANQEIRYSEVTPYH (SEQ ID NO: 113). In some embodiments, the polypeptide does not comprise the amino acid sequence of NQTN (SEQ ID NO: 119). In some embodiments, the polypeptide does not comprise the contiguous amino acid sequence of NQTN (SEQ ID NO: 119). In some embodiments, the polypeptide does not comprise the contiguous amino acid sequence of NQTN (SEQ ID NO: 119) at the C-terminus of the polypeptide. As used herein, the C-terminus of the polypeptide in reference to the contiguous amino acid sequence of NQTN (SEQ ID NO: 119) means the C-terminus of the protease, which may be N-terminal to a linker or another polypeptide if the protease is linked to another polypeptide, such as the Fc domains provided for herein. In some embodiments, the polypeptide comprises the sequence of QQTN (SEQ ID NO: 120) at the C-terminus of the polypeptide instead of the sequence of NQTN (SEQ ID NO: 119).

[0378] -65-

[0379] IPTS / 200163498.2 DOCKET NO. SES-020WO PATENT

[0380] In some embodiments, the polypeptide has a cysteine (C) at the position in said polypeptide which corresponds to position 94 of SEQ ID NO: 35. In some embodiments, the polypeptide has, at the positions in said variant sequence which correspond to positions 84, 262, 284 and 286 of SEQ ID NO: 35, a lysine (K), a histidine (H), an aspartic acid (D) and an aspartic acid (D), respectively. In some embodiments, the polypeptide has Ig protease activity. In some embodiments, the polypeptide has IgG protease activity. In some embodiments, the polypeptide is wherein is less immunogenic than an IdeS polypeptide, such as an IdeS polypeptide that comprises the amino acid sequence of SEQ ID NO: 35, SEQ ID NO: 36 or SEQ ID NO: 171. In some embodiments, the polypeptide produces greater quantity of IgG cleavage fragments than IdeS (e.g., SEQ ID NO: 35, SEQ ID NO: 36, or SEQ ID NO: 171). In some embodiments, the polypeptide has a positively charged amino acid at the position in said variant which corresponds to position 130 of SEQ ID NO: 35, such as, arginine (R) or lysine (K). In some embodiments, the polypeptide has a positively charged amino acid at the position in said variant which corresponds to position 131 of SEQ ID NO: 35, such as arginine (R) or lysine (K).

[0381] In some embodiments, the polypeptide comprises a mutation at position 163, wherein the position corresponds to the numbering as set forth in SEQ ID NO: 35. In some embodiments, the polypeptide having a mutation at position 163 exhibits improved physical properties. In some embodiments, the polypeptide having a mutation at position 163 exhibits improved manufacturability. In some embodiments, the polypeptide having a mutation at position 163 exhibits improved aggregation, thermostability, self-association, polyreactivity, and binding to pre-existing ADA. In some embodiments, the polypeptide, such as those provided for herein, having a mutation at position 163 exhibits improved aggregation, thermostability, selfassociation, polyreactivity, and binding to pre-existing ADA. Accordingly, in some embodiments, the polypeptide having the amino acid sequence of any one of SEQ ID NO: 58, 59, or 60 exhibits improved physical properties. In some embodiments, the polypeptide having the amino acid sequence of any one of SEQ ID NO: 58, 59, or 60 exhibits improved manufacturability. In some embodiments, the polypeptide having the amino acid sequence of any one of SEQ ID NO: 58, 59, or 60 exhibits improved aggregation, thermostability, selfassociation, polyreactivity, and binding to pre-existing ADA.

[0382] In some embodiments, the polypeptide is linked to a protease resistant Fc domain. In some embodiments, the protease resistant Fc domain is linked to the N-terminus or the C-

[0383] -66-

[0384] IPTS / 200163498.2 DOCKET NO. SES-020WO PATENT terminus of the polypeptide. In some embodiments, the protease resistant Fc domain is linked to the N-terminus of the polypeptide. In some embodiments, the protease resistant Fc domain is linked to the C-terminus of the polypeptide. In some embodiments, the protease resistant Fc domain is linked to the polypeptide through a linker. In some embodiments, the protease resistant Fc domain is linked to the polypeptide through a peptide linker. In some embodiments, the peptide linker is a charged peptide linker. As used herein, “charged peptide linker” is meant to refer to a peptide linker having at least one positively charged amino acid residue, such as, but not limited to, a lysine (K), an arginine (R), or a histidine (H). In some embodiments, the protease resistant Fc domain is as provided for herein. In some embodiments, the protease resistant Fc domain comprises the amino acid sequence of any one of SEQ ID NOs: 11, 12, 13, 19, 20, 21, 22, 23, 24, 25, 42, 43, 44, 45, 46, 47, 48, 49, 200, 201, or 202. In some embodiments, the protease resistant Fc domain comprises the amino acid sequence of SEQ ID NO: 23. In some embodiments, the polypeptide comprises a polypeptide from N-terminus to C-terminus a protease resistant Fc domain, a peptide linker, and a polypeptide, wherein the C-terminus of the protease resistant Fc domain is linked to the N-terminus of the peptide linker and the C-terminus of the peptide linker is linked to the N-terminus of the polypeptide.

[0385] In some embodiments, the polypeptide comprises the amino acid sequence as set forth in Table 7 in the column labeled “Full sequence” of VRT-1 through VRT-4. In some embodiments, the polypeptide has Ig protease activity, such as IgG protease activity. In some embodiments, the polypeptide is less immunogenic than an IdeS polypeptide, wherein the IdeS polypeptide optionally comprises the amino acid sequence of SEQ ID NO: 35, SEQ ID NO: 36 or SEQ ID NO: 171. In some embodiments, the polypeptide produces greater quantity of IgG cleavage fragments than IdeS (e.g., SEQ ID NO: 35, SEQ ID NO: 36, or SEQ ID NO: 171). In some embodiments, the polypeptide of SEQ ID NO: 171 produces greater quantity of IgG cleavage fragments than IdeS.

[0386] In some embodiments, the polypeptide comprises any mutation, or set of mutation, as provided in PCT Publication Nos. WO2016128558, and WO2016128559, or U.S. Patent Nos. 10,696,959, 11,214,784, 10,758,597, or 11,524,057, each of which is hereby incorporated by reference in its entirety.

[0387] IdeS (Immunoglobulin G-degrading enzyme of Streptococcus pyogenes) is an enzyme derived from the bacterium Streptococcus pyogenes that cleaves IgG antibodies and has been

[0388] -67-

[0389] IPTS / 200163498.2 DOCKET NO. SES-020WO PATENT explored for its potential therapeutic applications, particularly in the field of transplantation immunology. The immunogenicity of IdeS is a significant consideration for its clinical use, as the human immune system can potentially recognize the enzyme as a foreign protein and generate an immune response. This response could lead to the production of anti-drug antibodies (AD As) that may neutralize the enzymatic activity of IdeS or alter its pharmacokinetics, potentially reducing its efficacy or increasing the risk of adverse effects. Furthermore, repeated administration of IdeS could enhance this immunogenic response, limiting its long-term utility in chronic conditions. Thus, for any given subject, the problems associated with the immunogenicity of IdeS are likely to present a barrier to the use of IdeS as a treatment. These problems may require increases to the dose of IdeS and / or preclude treatment with IdeS entirely, particularly if repeat administrations are required.

[0390] Existing approaches to problems of this type involve, for example, PEGylation of a therapeutic agent to reduce immunogenicity or co-admini strati on of the therapeutic agent with an immune-suppressive agent. Some other approaches involve identification of specific positions within the sequence of IdeS which, when modified, lead to polypeptides for which the problems associated with immunogenicity are reduced as compared to IdeS. Thus, some modifications may increase the efficacy at cleaving IgG of the polypeptide of the disclosure relative to IdeS, thereby indirectly reducing immunogenicity by permitting the use of a lower dose or concentration to achieve the same effect. Alternatively, or in addition, other modifications may directly reduce immunogenicity by reducing the ability of IdeS-specific antibodies to recognize the polypeptide of the disclosure relative to IdeS.

[0391] The present disclosure provides IdeS variants that have, amongst other properties, lower immunogenicity as compared to wild-type IdeS. This can enable the variants to be administered to patients more than one time. The variants provided herein, can also comprises mutations or changes as compared to the wild-type sequence that impact post-translational modifications, aggregation, glycosylation, impurities, and formulation components), reduce B cell epitopes, thereby reducing the likelihood for creating antibody epitopes that would be produce antibodies in vivo against the variants, reduce T cell epitopes (linear peptide sequences that can be displayed on MHCII and lead to de novo immune responses), and reduce the likelihood of ADA (anti-drug antibodies). Thus, in some embodiments, a polypeptide having an IgG protease activity has one more B cell epitopes removed. In some embodiments, a polypeptide having an IgG protease

[0392] -68-

[0393] IPTS / 200163498.2 DOCKET NO. SES-020WO PATENT activity has one or more T cell epitopes removed. Tn some embodiments, a polypeptide having an IgG protease activity does not have, or does not have significant, chemical liabilities. In some embodiments, a polypeptide having an IgG protease activity has one or more B cell epitopes removed, one more T cell epitopes removed, and does not have, or does not have significant, chemical liabilities. Without being bound to any particular theory, a polypeptide having an IgG protease activity, such as those provided herein, can be utilized in a way that can be administered more than once, and thus can be used in a non-dose limiting manner or method..

[0394] In some embodiments, the polypeptide having IgG protease activity is as effective, or more effective at cleaving IgG than the wild-type protease, and / or is less immunogenic as the wild-type protease. In some embodiments, the polypeptide having IgG protease activity is as effective at cleaving IgG as compared to the wild-type protease. In some embodiments, the polypeptide having IgG protease activity is more effective at cleaving IgG than the wild-type protease. In some embodiments, the polypeptide having IgG protease activity is less effective at cleaving IgG than the wild-type protease, but is also less immunogenic and / or has fewer chemical liabilities as compared to the wild-type protease. This properties can be as compared to the wild-type IdeS or the wild-type Ides that is fused to a Fc molecule, such as those provided for herein. In some embodiments, the polypeptide having IgG protease activity is less immunogenic than the wild-type protease. In some embodiments, the polypeptide having IgG protease activity is as immunogenic as the wild-type protease.

[0395] In some embodiments, the polypeptide having IgG protease activity is as effective, or more effective at cleaving IgG than the wild-type IdeS, and / or is less immunogenic, or as immunogenic, as the wild-type IdeS. In some embodiments, the polypeptide having IgG protease activity is as effective at cleaving IgG than the wild-type IdeS. In some embodiments, the polypeptide having IgG protease activity is more effective at cleaving IgG than the wild-type IdeS. In some embodiments, the polypeptide having IgG protease activity is less immunogenic than the wild-type IdeS. In some embodiments, the polypeptide having IgG protease activity is as immunogenic as the wild-type IdeS.

[0396] In some embodiments, PRT-1 through PRT-4 is as effective, or more effective at cleaving IgG than the wild-type protease, and / or is less immunogenic, or as immunogenic, as the wild-type protease. In some embodiments, PRT-1 through PRT-4 is as effective at cleaving IgG than the wild-type protease. In some embodiments, PRT-1 through PRT-4 is more effective at

[0397] -69-

[0398] IPTS / 200163498.2 DOCKET NO. SES-020WO PATENT cleaving IgG than the wild-type protease. In some embodiments, PRT-1 through PRT-4 is less immunogenic than the wild-type protease. In some embodiments, PRT-1 through PRT-4 is as immunogenic as the wild-type protease.

[0399] In some embodiments, PRT-1 through PRT-4 is as effective, or more effective at cleaving IgG than the wild-type protease, and / or is less immunogenic, or as immunogenic, as the wild-type IdeS. In some embodiments, PRT-1 through PRT-4 is as effective at cleaving IgG than the wild-type IdeS. In some embodiments, PRT-1 through PRT-4 is more effective at cleaving IgG than the wild-type IdeS. In some embodiments, PRT-1 through PRT-4 is less immunogenic than the wild-type IdeS. In some embodiments, PRT-1 through PRT-4 is as immunogenic as the wild-type IdeS.

[0400] In some embodiments, VRT-1 through VRT-4 is as effective, or more effective at cleaving IgG than the wild-type protease, and / or is less immunogenic, or as immunogenic, as the wild-type protease. In some embodiments, VRT-1 through VRT-4 is as effective at cleaving IgG than the wild-type protease. In some embodiments, VRT-1 through VRT-4 is more effective at cleaving IgG than the wild-type protease. In some embodiments, VRT-1 through VRT-4 is less immunogenic than the wild-type protease. In some embodiments, VRT-1 through VRT-4 is as immunogenic as the wild-type protease.

[0401] In some embodiments, VRT-1 through VRT-4 is as effective, or more effective at cleaving IgG than the wild-type protease, and / or is less immunogenic, or as immunogenic, as the wild-type IdeS. In some embodiments, VRT-1 through VRT-4 is as effective at cleaving IgG than the wild-type IdeS. In some embodiments, VRT-1 through VRT-4 is more effective at cleaving IgG than the wild-type IdeS. In some embodiments, VRT-1 through VRT-4 is less immunogenic than the wild-type IdeS. In some embodiments, VRT-1 through VRT-4 is as immunogenic as the wild-type IdeS.

[0402] Fc Fusion Molecules

[0403] Provided herein are therapeutic compounds, e.g., therapeutic protein molecules, e.g., fusion proteins, including an effector binding / modulating moiety and a polypeptide having protease activity. Also provided are methods of using and making the therapeutic compounds. In some embodiments, the effector binding / modulating moiety is a Fc polypeptide, such as, but not limited to, those provided herein.

[0404] -70-

[0405] IPTS / 200163498.2 DOCKET NO. SES-020WO PATENT

[0406] The disclosure provides for, for example, a polypeptide comprising a polypeptide having protease activity and a Fc polypeptide domain, wherein the polypeptide is covalently or non- covalently connected to the Fc polypeptide domain. In some embodiments, the Fc polypeptide domain is a Fc polypeptide, such as those provided herein. Accordingly, in some embodiments, a polypeptide comprising a polypeptide having protease activity and a Fc polypeptide domain is provided, wherein the polypeptide is covalently or non-covalently connected to the Fc polypeptide, is provided. The polypeptide having protease activity can be fused or linked (i.e., conjugated) to the Fc polypeptide. In some embodiments, the polypeptide having protease activity is non-covalently associated with the Fc polypeptide domain.

[0407] In some embodiments, the Fc polypeptide domain is an IgG Fc polypeptide, such as a Fc polypeptide that is, or derived from an IgGi, IgG2, IgGs, or IgG4 Fc polypeptide. In some embodiments, the Fc polypeptide domain is a Fc polypeptide domain, such as the variants provided for herein. In some embodiments, the Fc polypeptide domain is protease resistant. The Fc polypeptide can be resistant to protease to which it is connected to, thus resistant to protease cleavage and / or binding by the protease. In some embodiments, the Fc polypeptide, such as those provided herein, comprises a mutation that renders the IgG Fc polypeptide resistant to cleavage by a protease, or a variant thereof. In some embodiments, the Fc polypeptide, such as those provided herein, comprises a mutation that renders the IgG Fc polypeptide resistant to binding by a protease, or a variant thereof. In some embodiments, the Fc polypeptide, such as those provided herein, comprises a set of mutations that render the IgG Fc polypeptide resistant to cleavage by a protease, or a variant thereof. In some embodiments, the Fc polypeptide, such as those provided herein, comprises a set of mutations that render the IgG Fc polypeptide resistant to binding by a protease, or a variant thereof. In some embodiments, the Fc polypeptide, such as those provided herein, comprises a mutation that renders the IgG Fc polypeptide resistant to cleavage by a protease, or a variant thereof, and / or binding by a protease, or a variant thereof. In some embodiments, the Fc polypeptide, such as those provided herein, comprises a set of mutations that render the IgG Fc polypeptide resistant to cleavage by a protease, or a variant thereof, and / or binding by a protease, or a variant thereof. In some embodiments, the Fc polypeptide, such as those provided herein, is resistant to cleavage by a protease, or a variant thereof. In some embodiments, the Fc polypeptide, such as those provided herein, is resistant to binding by a protease, or a variant thereof. In some embodiments, the Fc polypeptide, such as

[0408] -71-

[0409] IPTS / 200163498.2 DOCKET NO. SES-020WO PATENT those provided herein, is resistant to cleavage by a protease, or a variant thereof, and / or binding by a protease, or a variant thereof. It can also be resistant to other proteases that the Fc polypeptide domain is not connected to. Fc polypeptides can self-associate with one another to form a dimer. The dimer can be a homodimer where the Fc polypeptide is the same in each polypeptide molecule or can be a heterodimer where the Fc polypeptide is different in each polypeptide molecule that forms the dimer.

[0410] In some embodiments, the Fc polypeptide is covalently or non-covalently conjugated to a polypeptide having IgG protease activity, wherein the polypeptide having IgG protease activity can be conjugated to N-terminus or C-terminus of the Fc polypeptide. In some embodiments, the Fc polypeptide is covalently or non-covalently conjugated to a polypeptide having IgG protease activity, wherein the polypeptide having IgG protease activity can be conjugated to the N- terminus of the Fc polypeptide. In some embodiments, the Fc polypeptide is covalently or non- covalently conjugated to a polypeptide having IgG protease activity, wherein the polypeptide having IgG protease activity can be conjugated to the C-terminus of the Fc polypeptide.

[0411] The Fc polypeptide and the polypeptide having protease activity can be physically tethered, covalently or non-covalently, directly or through a linker entity, to one another, e.g., as a domain of the same linear primary amino acid sequence in a single polypeptide. The polypeptides can associate with one another through the Fc polypeptide, such that a dimer is created having a first and second polypeptide. In some embodiments, the first polypeptide comprises a polypeptide having protease activity linked to a Fc polypeptide domain,. In some embodiments, the second polypeptide comprises a Fc polypeptide domain, such as those provided for herein. In some embodiments, the second polypeptide does not comprise a protease domain. In some embodiments, the second polypeptide comprises a protease domain.

[0412] This multi-domain molecule can be referred to as a therapeutic protein molecule. In some embodiments, the protease and the IgG Fc molecule are provided in a therapeutic protein molecule, e.g., a fusion protein.

[0413] In some embodiments, a therapeutic compound or compound comprises a polypeptide comprising a polypeptide having protease activity covalently or non-covalently conjugated to a Fc polypeptide. In some embodiments, a therapeutic molecule comprises a fusion protein comprising a polypeptide having protease activity fused, e.g., directly or through a linking moiety comprising one or more amino acid residues, to a Fc polypeptide. In some embodiments,

[0414] IPTS / 200163498.2 DOCKET NO. SES-020WO PATENT a therapeutic compound or compound comprises a polypeptide comprising a polypeptide having protease activity linked by a non-covalent bond or a covalent bond, e.g., a covalent bond other than a peptide bond, e.g., a sulfhydryl bond, to a Fc polypeptide.

[0415] In some embodiments, the protease is an IgG cleaving protease. In some embodiments, the IgG cleaving protease is selected from IdeS, IdeSsuis, IdeE, IdeZ, IdeE2, IdeZ2, Ide85, and IdeC. In some embodiments, the IgG cleaving protease is IdeS. In some embodiments, the IgG cleaving protease is IdeSsuis. In some embodiments, the IgG cleaving protease is IdeE. In some embodiments, the IgG cleaving protease is IdeZ. In some embodiments, the IgG cleaving protease is IdeE2. In some embodiments, the IgG cleaving protease is IdeZ2. In some embodiments, the IgG cleaving protease is Ide85. In some embodiments, the IgG cleaving protease is IdeC. The IgG protease can be a variant protease of any of the foregoing. Examples of IgG proteases and variant proteases include, but are not limited to, those that are described in W02003051914, WO2006131347, WO2016128558, WO2016128559, WO2016012285, W02008071418, WO2012119983, W02010057626, WO2015184325, WO2021026264, WO2021021989, WO2018034346, W02010089126, W0200908027, WO2008136735, WO2015181356, WO2018093868, WO2013037824, WO2017134274, W02016046220, W02015040125, W02009033670, W02004096157, WO2010123885, WO2010118337, W02019075360, W02007019376, and U.S. Patent No. 10,836,815, each of which are hereby incorporated by reference in their entirety. Non-limiting examples of proteases include, but are not limited to proteases comprise an amino acid sequence having at least 50%, 60%, 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%, or 100% sequence identity to, or is identical to, a sequence provided in Table 4, or Table 5.

[0416] In some embodiments, a polypeptide is covalently or non-covalently conjugated to the N- terminus of a Fc polypeptide. In some embodiments, a polypeptide is covalently or non- covalently conjugated to the C-terminus of a Fc polypeptide. In some embodiments, there can be a linker moiety, such as a peptide moiety or a non-peptide linkage, between the polypeptide and the N-terminus or the C-terminus of the Fc polypeptide.

[0417] In some embodiments, a polypeptide is covalently or non-covalently conjugated to a nanobody and a Fc polypeptide, wherein the C-terminus of the polypeptide is covalently or non- covalently conjugated to the N-terminus of a nanobody, and the N-terminus of the polypeptide is

[0418] IPTS / 200163498.2 DOCKET NO. SES-020WO PATENT covalently or non-covalently conjugated to the C-terminus of a Fc polypeptide. In some embodiments, a polypeptide is covalently or non-covalently conjugated to a nanobody and a Fc polypeptide, wherein the N-terminus of a polypeptide is covalently or non-covalently conjugated to the C-terminus of a nanobody, and the C-terminus of the polypeptide is covalently or non- covalently conjugated to the N-terminus of a Fc polypeptide. In some embodiments, the polypeptide is as provided herein. In some embodiments, the variant IgG Fc is as provided herein. In some embodiments, the nanobody is as provided herein.

[0419] In some embodiments, a polypeptide comprising a polypeptide having protease activity covalently or non-covalently conjugated to a Fc polypeptide further comprises a tag, such as a purification tag or detection tag. The tag can be used to facilitate the purification, isolation, or detection of the polypeptide. A non-limiting example of such a tag is a histidine tag, which is a plurality of hisitidines (e g., 6 or more histidines). In some embodiments, a polypeptide comprising a polypeptide having protease activity covalently or non-covalently conjugated to a Fc polypeptide further comprises a histidine tag, wherein the C-terminus of the polypeptide having protease activity is covalently or non-covalently conjugated to the N-terminus of a Fc polypeptide, and wherein the C-terminus of the Fc polypeptide is covalently or non-covalently conjugated to the N-terminus of the tag. In some embodiments, a polypeptide comprising a polypeptide having protease activity covalently or non-covalently conjugated to a nanobody and a Fc polypeptide further comprises a tag. In some embodiments, a polypeptide comprising a polypeptide having protease activity covalently or non-covalently conjugated to a nanobody and a Fc polypeptide further comprises a tag, wherein the N-terminus of the polypeptide having protease activity is covalently or non-covalently conjugated to the C-terminus of the nanobody, wherein the C-terminus of the polypeptide having protease activity is covalently or non- covalently conjugated to the N-terminus of a Fc polypeptide, and wherein the C-terminus of the Fc polypeptide is covalently or non-covalently conjugated to the N-terminus of the tag. In some embodiments, a polypeptide comprising a polypeptide having protease activity covalently or non-covalently conjugated to a nanobody and a Fc polypeptide further comprises a histidine tag, wherein the N-terminus of the polypeptide having protease activity is covalently or non- covalently conjugated to the C-terminus of a Fc polypeptide, wherein the C-terminus of the polypeptide having protease activity is covalently or non-covalently conjugated to the N-

[0420] IPTS / 200163498.2 DOCKET NO. SES-020WO PATENT terminus of the nanobody, and wherein the C-terminus of the nanobody is covalently or non- covalently conjugated to the N-terminus of the tag.

[0421] In some embodiments, the Fc polypeptide comprises an amino acid sequence having at least 50%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%, or 100% sequence identity, or is identical, to a sequence provided in Table 3. In some embodiments, a polypeptide is covalently or non-covalently conjugated to a Fc polypeptide, wherein the Fc polypeptide comprises an amino acid sequence having at least 50%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%, or 100% sequence identity, or is identical to SEQ ID NO: 11, 12, 13, 19, 20, 21, 22, 23, 24, 25, 42, 43, 44, 45, 46, 47, 48, 49, 200, 201, or 202. In some embodiments, a polypeptide is covalently or non- covalently conjugated to a Fc polypeptide, wherein the Fc polypeptide comprises an amino acid sequence having at least 50%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%, or 100% sequence identity to, or is identical to, SEQ ID NO: 23.

[0422] As used herein, the positions referenced in the Fc polypeptides as positions for a mutation refer to EU numbering system. The Fc polypeptides can aligned with a wild-type sequence to determine the positions that are mutated according to the numbering system.

[0423] As used herein in reference to a polypeptide that has a % identity to a reference sequence and further comprises a mutation at one or more positions, means a polypeptide that has the recited % identity to the reference sequence and also has one or more of the mutations at the recited positions.

[0424] In some embodiments, a polypeptide is covalently or non-covalently conjugated to a Fc polypeptide, wherein the Fc polypeptide comprises an amino acid sequence of SEQ ID NO: 11, 12, 13, 19, 20, 21, 22, 23, 24, 25, 42, 43, 44, 45, 46, 47, 48, 49, 200, 201, or 202. In some embodiments, a polypeptide is covalently or non-covalently conjugated to a Fc polypeptide, wherein the Fc polypeptide comprises an amino acid sequence of SEQ ID NO: 11. In some embodiments, a polypeptide is covalently or non-covalently conjugated to a Fc polypeptide, wherein the Fc polypeptide comprises an amino acid sequence of SEQ ID NO: 12. In some embodiments, a polypeptide is covalently or non-covalently conjugated to a Fc polypeptide,

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[0426] IPTS / 200163498.2 DOCKET NO. SES-020WO PATENT wherein the Fc polypeptide comprises an amino acid sequence of SEQ ID NO: 13. Tn some embodiments, a polypeptide is covalently or non-covalently conjugated to a Fc polypeptide, wherein the Fc polypeptide comprises an amino acid sequence of SEQ ID NO: 19. In some embodiments, a polypeptide is covalently or non-covalently conjugated to a Fc polypeptide, wherein the Fc polypeptide comprises an amino acid sequence of SEQ ID NO: 20. In some embodiments, a polypeptide is covalently or non-covalently conjugated to a Fc polypeptide, wherein the Fc polypeptide comprises an amino acid sequence of SEQ ID NO: 21. In some embodiments, a polypeptide is covalently or non-covalently conjugated to a Fc polypeptide, wherein the Fc polypeptide comprises an amino acid sequence of SEQ ID NO: 22. In some embodiments, a polypeptide is covalently or non-covalently conjugated to a Fc polypeptide, wherein the Fc polypeptide comprises an amino acid sequence of SEQ ID NO: 23. In some embodiments, a polypeptide is covalently or non-covalently conjugated to a Fc polypeptide, wherein the Fc polypeptide comprises an amino acid sequence of SEQ ID NO: 24. In some embodiments, a polypeptide is covalently or non-covalently conjugated to a Fc polypeptide, wherein the Fc polypeptide comprises an amino acid sequence of SEQ ID NO: 25. In some embodiments, a polypeptide is covalently or non-covalently conjugated to a Fc polypeptide, wherein the Fc polypeptide comprises an amino acid sequence of SEQ ID NO: 42. In some embodiments, a polypeptide is covalently or non-covalently conjugated to a Fc polypeptide, wherein the Fc polypeptide comprises an amino acid sequence of SEQ ID NO: 43. In some embodiments, a polypeptide is covalently or non-covalently conjugated to a Fc polypeptide, wherein the Fc polypeptide comprises an amino acid sequence of SEQ ID NO: 44. In some embodiments, a polypeptide is covalently or non-covalently conjugated to a Fc polypeptide, wherein the Fc polypeptide comprises an amino acid sequence of SEQ ID NO: 45. In some embodiments, a polypeptide is covalently or non-covalently conjugated to a Fc polypeptide, wherein the Fc polypeptide comprises an amino acid sequence of SEQ ID NO: 46. In some embodiments, a polypeptide is covalently or non-covalently conjugated to a Fc polypeptide, wherein the Fc polypeptide comprises an amino acid sequence of SEQ ID NO: 47. In some embodiments, a polypeptide is covalently or non-covalently conjugated to a Fc polypeptide, wherein the Fc polypeptide comprises an amino acid sequence of SEQ ID NO: 48. In some embodiments, a polypeptide is covalently or non-covalently conjugated to a Fc polypeptide, wherein the Fc polypeptide comprises an amino acid sequence of SEQ ID NO: 49. In some

[0427] -76-

[0428] IPTS / 200163498.2 DOCKET NO. SES-020WO PATENT embodiments, a polypeptide is covalently or non-covalently conjugated to a Fc polypeptide, wherein the Fc polypeptide comprises an amino acid sequence of SEQ ID NO: 200. In some embodiments, a polypeptide is covalently or non-covalently conjugated to a Fc polypeptide, wherein the Fc polypeptide comprises an amino acid sequence of SEQ ID NO: 201. In some embodiments, a polypeptide is covalently or non-covalently conjugated to a Fc polypeptide, wherein the Fc polypeptide comprises an amino acid sequence of SEQ ID NO: 202.

[0429] In some embodiments, a polypeptide is covalently or non-covalently conjugated to a Fc polypeptide, wherein the Fc polypeptide comprises an amino acid sequence having a C-terminal lysine (K). In some embodiments, a polypeptide is covalently or non-covalently conjugated to a Fc polypeptide, wherein the Fc polypeptide comprises an amino acid sequence that does not have (comprise) a C-terminal lysine (K).

[0430] In some embodiments, the protease present in the polypeptides provided for herein comprise an amino acid sequence having at least 50%, at least 60%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%, or 100% sequence identity to, or is identical to, a sequence provided in Table 5, wherein the protease is covalently or non- covalently conjugated to a Fc polypeptide, and wherein the Fc polypeptide comprises an amino acid sequence having at least 50%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%, or 100% sequence identity to any sequence provided in Table 3.

[0431] In some embodiments, a polypeptide comprises an amino acid sequence having at least 50%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%, or 100% sequence identity to SEQ ID NO: 57, 58, 59, 60, 189, 190, 191, 192, 193, 194, 195, 196, 197, 198, or 199, wherein the polypeptide is covalently or non-covalently conjugated to a Fc polypeptide, and wherein the Fc polypeptide comprises an amino acid sequence having at least 50%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%, or 100% sequence identity to SEQ ID NO: 11, 12, 13, 19, 20, 21, 22, 23, 24, 25, 42, 43, 44, 45, 46, 47, 48, 49, 200, 201, or 202.

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[0433] IPTS / 200163498.2 DOCKET NO. SES-020WO PATENT

[0434] In some embodiments, a polypeptide comprises an amino acid sequence having at least 50%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%, or 100% sequence identity to SEQ ID NO: 57, 58, 59, 60, 189, 190, 191, 192, 193, 194, 195, 196, 197, 198, or 199, wherein the polypeptide is covalently or non-covalently conjugated to a Fc polypeptide, and the Fc polypeptide comprises an amino acid sequence having at least 50%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%, or 100% sequence identity to SEQ ID NO: 23.

[0435] In some embodiments, a polypeptide is covalently or non-covalently conjugated to a Fc polypeptide, wherein the Fc polypeptide comprises an amino acid sequence as provided in Table 3. In some embodiments, a polypeptide is covalently or non-covalently conjugated to a Fc polypeptide, wherein the Fc polypeptide comprises an amino acid sequence of SEQ ID NO: 11, 12, 13, 19, 20, 21, 22, 23, 24, 25, 42, 43, 44, 45, 46, 47, 48, 49, 200, 201, or 202. In some embodiments, a polypeptide is covalently or non-covalently conjugated to a Fc polypeptide, wherein the Fc polypeptide comprises an amino acid sequence of SEQ ID NO: 11. In some embodiments, a polypeptide is covalently or non-covalently conjugated to a Fc polypeptide, wherein the Fc polypeptide comprises an amino acid sequence of SEQ ID NO: 12. In some embodiments, a polypeptide is covalently or non-covalently conjugated to a Fc polypeptide, wherein the Fc polypeptide comprises an amino acid sequence of SEQ ID NO: 13. In some embodiments, a polypeptide is covalently or non-covalently conjugated to a Fc polypeptide, wherein the Fc polypeptide comprises an amino acid sequence of SEQ ID NO: 19. In some embodiments, a polypeptide is covalently or non-covalently conjugated to a Fc polypeptide, wherein the Fc polypeptide comprises an amino acid sequence of SEQ ID NO: 20. In some embodiments, a polypeptide is covalently or non-covalently conjugated to a Fc polypeptide, wherein the Fc polypeptide comprises an amino acid sequence of SEQ ID NO: 21. In some embodiments, a polypeptide is covalently or non-covalently conjugated to a Fc polypeptide, wherein the Fc polypeptide comprises an amino acid sequence of SEQ ID NO: 22. In some embodiments, a polypeptide is covalently or non-covalently conjugated to a Fc polypeptide, wherein the Fc polypeptide comprises an amino acid sequence of SEQ ID NO: 23. In some embodiments, a polypeptide is covalently or non-covalently conjugated to a Fc polypeptide,

[0436] -78-

[0437] IPTS / 200163498.2 DOCKET NO. SES-020WO PATENT wherein the Fc polypeptide comprises an amino acid sequence of SEQ ID NO: 24. Tn some embodiments, a polypeptide is covalently or non-covalently conjugated to a Fc polypeptide, wherein the Fc polypeptide comprises an amino acid sequence of SEQ ID NO: 25. In some embodiments, a polypeptide is covalently or non-covalently conjugated to a Fc polypeptide, wherein the Fc polypeptide comprises an amino acid sequence of SEQ ID NO: 42. In some embodiments, a polypeptide is covalently or non-covalently conjugated to a Fc polypeptide, wherein the Fc polypeptide comprises an amino acid sequence of SEQ ID NO: 43. In some embodiments, a polypeptide is covalently or non-covalently conjugated to a Fc polypeptide, wherein the Fc polypeptide comprises an amino acid sequence of SEQ ID NO: 44. In some embodiments, a polypeptide is covalently or non-covalently conjugated to a Fc polypeptide, wherein the Fc polypeptide comprises an amino acid sequence of SEQ ID NO: 45. In some embodiments, a polypeptide is covalently or non-covalently conjugated to a Fc polypeptide, wherein the Fc polypeptide comprises an amino acid sequence of SEQ ID NO: 46. In some embodiments, a polypeptide is covalently or non-covalently conjugated to a Fc polypeptide, wherein the Fc polypeptide comprises an amino acid sequence of SEQ ID NO: 47. In some embodiments, a polypeptide is covalently or non-covalently conjugated to a Fc polypeptide, wherein the Fc polypeptide comprises an amino acid sequence of SEQ ID NO: 48. In some embodiments, a polypeptide is covalently or non-covalently conjugated to a Fc polypeptide, wherein the Fc polypeptide comprises an amino acid sequence of SEQ ID NO: 49. In some embodiments, a polypeptide is covalently or non-covalently conjugated to a Fc polypeptide, wherein the Fc polypeptide comprises an amino acid sequence of SEQ ID NO: 200. In some embodiments, a polypeptide is covalently or non-covalently conjugated to a Fc polypeptide, wherein the Fc polypeptide comprises an amino acid sequence of SEQ ID NO: 201. In some embodiments, a polypeptide is covalently or non-covalently conjugated to a Fc polypeptide, wherein the Fc polypeptide comprises an amino acid sequence of SEQ ID NO: 202.

[0438] In some embodiments, a polypeptide comprises any sequence provided in Table 5, wherein the polypeptide is covalently or non-covalently conjugated to a Fc polypeptide, wherein the Fc polypeptide comprises an amino acid sequence as provided in Table 3. In some embodiments, a polypeptide comprises an amino acid sequence of SEQ ID NO: 57, 58, 59, 60, 189, 190, 191, 192, 193, 194, 195, 196, 197, 198, or 199, wherein the polypeptide is covalently or non-covalently conjugated to a Fc polypeptide, wherein the Fc polypeptide comprises an

[0439] -79-

[0440] IPTS / 200163498.2 DOCKET NO. SES-020WO PATENT amino acid sequence of SEQ ID NO: 11 , 12, 13, 19, 20, 21, 22, 23, 24, 25, 42, 43, 44, 45, 46, 47, 48, 49, 200, 201, or 202.

[0441] In some embodiments, a polypeptide comprises an amino acid sequence of SEQ ID NO: 57, 58, 59, 60, 189, 190, 191, 192, 193, 194, 195, 196, 197, 198, or 199, wherein the polypeptide is covalently or non-covalently conjugated to an Fc polypeptide, wherein the Fc polypeptide comprises an amino acid sequence of SEQ ID NO: 23. In some embodiments, a polypeptide comprises an amino acid sequence of SEQ ID NO: 57, 58, 59, 60, 189, 190, 191, 192, 193, 194, 195, 196, 197, 198, or 199, wherein the polypeptide is conjugated via a linker, such as those provided herein, to an Fc polypeptide having an amino acid sequence of SEQ ID NO: 23. In some embodiments, a polypeptide comprises an amino acid sequence of SEQ ID NO: 57, 58, 59, 60, 189, 190, 191, 192, 193, 194, 195, 196, 197, 198, or 199, wherein the polypeptide is conjugated via a linker, such as a glycine / serine linker, to an Fc polypeptide having an amino acid sequence of SEQ ID NO: 23. In some embodiments, a polypeptide comprises an amino acid sequence of SEQ ID NO: 57, 58, 59, 60, 189, 190, 191, 192, 193, 194, 195, 196, 197, 198, or 199, wherein the polypeptide is conjugated via a linker having the amino acid sequence of GGGSKGGGSK (SEQ ID NO: 121), to an Fc polypeptide having an amino acid sequence of SEQ ID NO: 23.

[0442] In some embodiments, a polypeptide is covalently or non-covalently conjugated to a Fc polypeptide, wherein the Fc polypeptide comprises an amino acid sequence having a C-terminal lysine (K).

[0443] In some embodiments, a polypeptide is covalently or non-covalently conjugated to a Fc polypeptide, wherein the Fc polypeptide comprises an amino acid sequence not having a C- terminal lysine (K).

[0444] In some embodiments, a polypeptide has the formula from N-terminus to C-terminus: Pl — variant Fc, wherein,

[0445] Pl comprises a polypeptide having protease activity; and variant Fc is any variant IgG Fc provided herein.

[0446] In some embodiments, a polypeptide has the formula from N-terminus to C-terminus: variant Fc — Pl, wherein,

[0447] -80-

[0448] IPTS / 200163498.2 DOCKET NO. SES-020WO PATENT

[0449] P1 comprises a polypeptide having protease activity; and variant Fc is any variant IgG Fc provided herein.

[0450] In some embodiments, a polypeptide has the formula from N-terminus to C-terminus: Pl — variant Fc, wherein,

[0451] Pl comprises a polypeptide having protease activity; variant Fc is any variant IgG Fc provided herein; and optionally wherein the protease is a glycosylation-resistant protease.

[0452] In some embodiments, a polypeptide has the formula from N-terminus to C-terminus: variant Fc — Pl, wherein,

[0453] Pl comprises a polypeptide having protease activity; variant Fc is any variant IgG Fc provided herein; and optionally wherein the protease is a glycosylation-resistant protease.

[0454] In some embodiments, Pl comprises a polypeptide having protease activity comprising an amino acid sequence having at least 50%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least

[0455] 96%, at least 97%, at least 98%, or at least 99%, or 100% sequence identity to any sequence provided in Table 5. In some embodiments, Pl comprises a polypeptide having protease activity comprising an amino acid sequence having at least 50%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%, or 100% sequence identity to SEQ ID NO: 57, 58, 59, 60, 189, 190, 191, 192, 193, 194, 195, 196, 197, 198, or 199.

[0456] In some embodiments, Pl comprises a polypeptide having protease activity comprising any sequence provided in Table 5. In some embodiments, P l comprises a polypeptide having protease activity comprising an amino acid sequence of SEQ ID NO: 57, 58, 59, 60, 189, 190, 191, 192, 193, 194, 195, 196, 197, 198, or 199.

[0457] In some embodiments, Pl comprises an IdeS, IdeSsuis, IdeE, IdeZ, IdeZ2, IdeE2, Ide85, or IdeC protease. In some embodiments, Pl comprises an IdeS, IdeSsuis, IdeE, IdeZ, IdeZ2, IdeE2, Ide85, or IdeC protease variant. In some embodiments, Pl comprises an IdeS, IdeSsuis, IdeE, IdeZ, IdeZ2, IdeE2, Ide85, or IdeC protease variant comprising an amino acid sequence

[0458] -81-

[0459] IPTS / 200163498.2 DOCKET NO. SES-020WO PATENT having at least 50%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%, or 100% sequence identity to SEQ ID NO: 57, 58, 59, 60, 189, 190, 191, 192, 193, 194, 195, 196, 197, 198, or 199.

[0460] In some embodiments, Pl comprises an IdeS, IdeSsuis, IdeE, IdeZ, IdeZ2, IdeE2, Ide85, or IdeC protease variant comprising an amino acid sequence of SEQ ID NO: 57, 58, 59, 60, 189, 190, 191, 192, 193, 194, 195, 196, 197, 198, or 199.

[0461] In some embodiments, the variant Fc comprises a Fc polypeptide. In some embodiments, the Fc polypeptide is such as those provided herein. In some embodiments, the Fc polypeptide comprises an amino acid sequence having at least 50%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%, or 100% sequence identity to SEQ ID NO: 11, 12, 13, 19, 20, 21, 22, 23, 24, 25, 42, 43, 44, 45, 46, 47, 48, 49, 200, 201, or 202. In some embodiments, the Fc polypeptide comprises an amino acid sequence having at least 50%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%, or 100% sequence identity to SEQ ID NO: 23.

[0462] In some embodiments, the Fc polypeptide comprises an amino acid sequence of SEQ ID NO: 11, 12, 13, 19, 20, 21, 22, 23, 24, 25, 42, 43, 44, 45, 46, 47, 48, 49, 200, 201, or 202. In some embodiments, the Fc polypeptide comprises an amino acid sequence of SEQ ID NO: 11. In some embodiments, the Fc polypeptide comprises an amino acid sequence of SEQ ID NO: 12. In some embodiments, the Fc polypeptide comprises an amino acid sequence of SEQ ID NO: 13. In some embodiments, the Fc polypeptide comprises an amino acid sequence of SEQ ID NO: 19. In some embodiments, the Fc polypeptide comprises an amino acid sequence of SEQ ID NO: 20. In some embodiments, the Fc polypeptide comprises an amino acid sequence of SEQ ID NO: 21. In some embodiments, the Fc polypeptide comprises an amino acid sequence of SEQ ID NO: 22. In some embodiments, the Fc polypeptide comprises an amino acid sequence of SEQ ID NO: 23. In some embodiments, the Fc polypeptide comprises an amino acid sequence of SEQ ID NO: 24. In some embodiments, the Fc polypeptide comprises an amino acid sequence of SEQ ID NO: 25. In some embodiments, the Fc polypeptide comprises an amino acid sequence of SEQ ID NO: 42. In some embodiments, the Fc polypeptide comprises an amino acid sequence of SEQ ID NO: 43. In

[0463] -82-

[0464] IPTS / 200163498.2 DOCKET NO. SES-020WO PATENT some embodiments, the Fc polypeptide comprises an amino acid sequence of SEQ ID NO: 44. In some embodiments, the Fc polypeptide comprises an amino acid sequence of SEQ ID NO: 45. In some embodiments, the Fc polypeptide comprises an amino acid sequence of SEQ ID NO: 46. In some embodiments, the Fc polypeptide comprises an amino acid sequence of SEQ ID NO: 47. In some embodiments, the Fc polypeptide comprises an amino acid sequence of SEQ ID NO: 48. In some embodiments, the Fc polypeptide comprises an amino acid sequence of SEQ ID NO: 49. In some embodiments, the Fc polypeptide comprises an amino acid sequence of SEQ ID NO: 200. In some embodiments, the Fc polypeptide comprises an amino acid sequence of SEQ ID NO: 201. In some embodiments, the Fc polypeptide comprises an amino acid sequence of SEQ ID NO: 202.

[0465] In some embodiments, a molecule comprises a dimer Fc molecule comprising a first Fc polypeptide having an amino acid sequence of SEQ ID NO: 23, and a second Fc polypeptide having an amino acid sequence of SEQ ID NO: 23, wherein the first Fc polypeptide and the second Fc polypeptide associate with each other to form the dimer Fc molecule; and a polypeptide having an amino acid sequence of any one, or any combination of SEQ ID NO: 57, 58, 59, 60, 189, 190, 191, 192, 193, 194, 195, 196, 197, 198, or 199, wherein the polypeptide is linked or conjugated to N- or C-terminus of the first Fc polypeptide and / or the second Fc polypeptide.

[0466] In some embodiments, a polypeptide comprises an amino acid sequence, and N-terminus to C-terminus orientation, as provided in Table 7. In some embodiments, the polypeptide comprises a protease moiety, wherein the protease moiety is a polypeptide having protease activity, such as those provided herein. In some embodiments, the polypeptide comprises an Fc moiety, wherein the Fc moiety is a Fc variant such as those provided herein. The Fc variant can also be modified to include mutations that extend the half-life. Examples of such mutations are provided for herein.

[0467] -83-

[0468] IPTS / 200163498.2 DOCKET NO. SES-020WO PATENT

[0469] -84-

[0470] IPTS / 200163498.2 DOCKET NO. SES-020WO PATENT

[0471] -85-

[0472] IPTS / 200163498.2 DOCKET NO. SES-020WO PATENT

[0473] -86-

[0474] IPTS / 200163498.2 DOCKET NO. SES-020WO PATENT

[0475] -87-

[0476] IPTS / 200163498.2 DOCKET NO. SES-020WO PATENT

[0477] -88-

[0478] IPTS / 200163498.2 DOCKET NO. SES-020WO PATENT

[0479] -89-

[0480] IPTS / 200163498.2 DOCKET NO. SES-020WO PATENT

[0481] -90-

[0482] IPTS / 200163498.2 DOCKET NO. SES-020WO PATENT

[0483] In some embodiments, a polypeptide comprises an amino acid sequence having at least 50%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%, or 100% sequence identity, or is identical, to any one amino acid sequence selected from SEQ ID NO: 114, 115, 116, 117, 118, 203, 204, 205, 206, 207, 208, 209, 210, 211, 212, 213, 214, 215, 216, or 217.

[0484] In some embodiments, a polypeptide comprises an amino acid sequence having at least 50%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%, or 100% sequence identity, or is identical, to the amino acid sequence of SEQ ID NO: 114. In some embodiments, a polypeptide comprises an amino acid sequence having at least 50%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%, or 100% sequence identity, or is identical, to the amino acid sequence of SEQ ID NO: 1 15. In some embodiments, a polypeptide comprises an amino acid sequence having at least 50%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%, or 100% sequence identity, or is identical, to the amino acid sequence of SEQ ID NO: 1 16. In some embodiments, a polypeptide comprises an amino acid sequence having at least 50%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%, or 100% sequence identity, or is identical, to the amino acid sequence of SEQ ID NO: 1 17. In some embodiments, a polypeptide comprises an amino acid sequence having at least 50%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%, or 100% sequence identity, or is identical, to the amino acid sequence of SEQ ID NO: 1 18. In some embodiments, a polypeptide comprises an amino acid sequence having at least 50%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%, or 100% sequence identity, or is identical, to the amino acid sequence of SEQ ID NO: 203. In some embodiments, a polypeptide comprises an amino acid sequence having at least 50%, at least

[0485] -91-

[0486] IPTS / 200163498.2 DOCKET NO. SES-020WO PATENT

[0487] 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%, or 100% sequence identity, or is identical, to the amino acid sequence of SEQ ID NO: 204. In some embodiments, a polypeptide comprises an amino acid sequence having at least 50%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%, or 100% sequence identity, or is identical, to the amino acid sequence of SEQ ID NO: 205. In some embodiments, a polypeptide comprises an amino acid sequence having at least 50%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%, or 100% sequence identity, or is identical, to the amino acid sequence of SEQ ID NO: 206. In some embodiments, a polypeptide comprises an amino acid sequence having at least 50%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%, or 100% sequence identity, or is identical, to the amino acid sequence of SEQ ID NO: 207. In some embodiments, a polypeptide comprises an amino acid sequence having at least 50%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%, or 100% sequence identity, or is identical, to the amino acid sequence of SEQ ID NO: 208. In some embodiments, a polypeptide comprises an amino acid sequence having at least 50%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%, or 100% sequence identity, or is identical, to the amino acid sequence of SEQ ID NO: 209. In some embodiments, a polypeptide comprises an amino acid sequence having at least 50%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%, or 100% sequence identity, or is identical, to the amino acid sequence of SEQ ID NO: 210. In some embodiments, a polypeptide comprises an amino acid sequence having at least 50%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%, or 100% sequence identity, or is identical, to the amino acid sequence of SEQ ID NO: 211. In some

[0488] -92-

[0489] IPTS / 200163498.2 DOCKET NO. SES-020WO PATENT embodiments, a polypeptide comprises an amino acid sequence having at least 50%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%, or 100% sequence identity, or is identical, to the amino acid sequence of SEQ ID NO: 212. In some embodiments, a polypeptide comprises an amino acid sequence having at least 50%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%, or 100% sequence identity, or is identical, to the amino acid sequence of SEQ ID NO: 213. In some embodiments, a polypeptide comprises an amino acid sequence having at least 50%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%, or 100% sequence identity, or is identical, to the amino acid sequence of SEQ ID NO: 214. In some embodiments, a polypeptide comprises an amino acid sequence having at least 50%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%, or 100% sequence identity, or is identical, to the amino acid sequence of SEQ ID NO: 215. In some embodiments, a polypeptide comprises an amino acid sequence having at least 50%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%, or 100% sequence identity, or is identical, to the amino acid sequence of SEQ ID NO: 216. In some embodiments, a polypeptide comprises an amino acid sequence having at least 50%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%, or 100% sequence identity, or is identical, to the amino acid sequence of SEQ ID NO: 217.

[0490] In some embodiments, the polypeptide has an amino acid sequence having at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to the amino acid sequence of SEQ ID NO: 114, provided that the polypeptide comprises a threonine (T) at position 60, an aspartic acid (D) at position 61, an isoleucine (I) at position 63, a tyrosine (Y) at position 64, a leucine (L) at position 69, a lysine (K) at position 74, a glycine (G) at position 76, a leucine (L) at position 85, a glutamic acid (E) at position 112, an

[0491] -93-

[0492] IPTS / 200163498.2 DOCKET NO. SES-020WO PATENT asparagine (N) at position 1 16, a lysine (K) at position 119, a lysine (K) at position 120, a tyrosine (Y) at position 121, a glycine (G) at position 130, a leucine (L) at position 134, an aspartic acid (D) at position 145, a serine (S) at position 146, a threonine (T) at position 148, an aspartic acid (D) at position 154, an aspartic acid (D) at position 190, an arginine (R) at position 198, an asparagine (N) at position 223, a tyrosine (Y) at position 225, a leucine (L) at position 227, a threonine (T) at position 231, a lysine (K) at position 241, an isoleucine (I) at position 251, a glycine (G) at position 261, an alanine (A) at position 273, a glutamic acid (E) at position 274, a glutamic acid (E) at position 278, a proline (P) at position 289, a leucine (L) at position 303, a glutamic acid (E) at position 314, and an isoleucine (I) at position 334, wherein the positions correspond to the numbering as set forth in SEQ ID NO: 35.

[0493] In some embodiments, the polypeptide has an amino acid sequence having at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to the amino acid sequence of SEQ ID NO: 115, provided that the polypeptide comprises a threonine (T) at position 60, an aspartic acid (D) at position 61, an isoleucine (I) at position 63, a tyrosine (Y) at position 64, a leucine (L) at position 69, a lysine (K) at position 74, a glycine (G) at position 76, a leucine (L) at position 85, a glutamic acid (E) at position 112, an asparagine (N) at position 116, a lysine (K) at position 119, a lysine (K) at position 120, a tyrosine (Y) at position 121, a glycine (G) at position 130, a leucine (L) at position 134, an aspartic acid (D) at position 145, a serine (S) at position 146, a threonine (T) at position 148, an aspartic acid (D) at position 154, an aspartic acid (D) at position 163, an aspartic acid (D) at position 190, an arginine (R) at position 198, an asparagine (N) at position 223, a tyrosine (Y) at position 225, a leucine (L) at position 227, a threonine (T) at position 231, a lysine (K) at position 241, an isoleucine (I) at position 251, a glycine (G) at position 261, an alanine (A) at position 273, a glutamic acid (E) at position 274, a glutamic acid (E) at position 278, a proline (P) at position 289, a leucine (L) at position 303, a glutamic acid (E) at position 314, and an isoleucine (I) at position 334, wherein the positions correspond to the numbering as set forth in SEQ ID NO: 35.

[0494] In some embodiments, the polypeptide has an amino acid sequence having at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%

[0495] -94-

[0496] IPTS / 200163498.2 DOCKET NO. SES-020WO PATENT sequence identity to the amino acid sequence of SEQ ID NO: 116, provided that the polypeptide comprises a threonine (T) at position 60, an aspartic acid (D) at position 61, an isoleucine (I) at position 63, a tyrosine (Y) at position 64, a leucine (L) at position 69, a lysine (K) at position 74, a glycine (G) at position 76, a leucine (L) at position 85, a glutamic acid (E) at position 112, an asparagine (N) at position 116, a lysine (K) at position 119, a lysine (K) at position 120, a tyrosine (Y) at position 121, a glycine (G) at position 130, a leucine (L) at position 134, an aspartic acid (D) at position 145, a serine (S) at position 146, a threonine (T) at position 148, an aspartic acid (D) at position 154, an asparagine (N) at position 163, an alanine (A) at position 166, an arginine (R) at position 167, a glutamine (Q) at position 168, an aspartic acid (D) at position 190, an arginine (R) at position 198, an asparagine (N) at position 223, a tyrosine (Y) at position 225, a leucine (L) at position 227, a threonine (T) at position 231, a lysine (K) at position 241, an isoleucine (I) at position 251, a glycine (G) at position 261, an alanine (A) at position 273, a glutamic acid (E) at position 274, a glutamic acid (E) at position 278, a proline (P) at position 289, a leucine (L) at position 303, a glutamic acid (E) at position 314, and an isoleucine (I) at position 334, wherein the positions correspond to the numbering as set forth in SEQ ID NO: 35.

[0497] In some embodiments, the polypeptide has an amino acid sequence having at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%>, at least 97%, at least 98%, or at least 99% sequence identity to the amino acid sequence of SEQ ID NO: 117, provided that the polypeptide comprises a threonine (T) at position 60, an aspartic acid (D) at position 61, an isoleucine (I) at position 63, a tyrosine (Y) at position 64, a leucine (L) at position 69, a lysine (K) at position 74, a glycine (G) at position 76, a leucine (L) at position 85, a glutamic acid (E) at position 112, an asparagine (N) at position 116, a lysine (K) at position 119, a lysine (K) at position 120, a tyrosine (Y) at position 121, a glycine (G) at position 130, a leucine (L) at position 134, an aspartic acid (D) at position 145, a serine (S) at position 146, a threonine (T) at position 148, an aspartic acid (D) at position 154, an asparagine (N) at position 163, an alanine (A) at position 166, an arginine (R) at position 167, a glutamine (Q) at position 168, an isoleucine (I) at position 169, an aspartic acid (D) at position 190, an arginine (R) at position 198, an asparagine (N) at position 223, a tyrosine (Y) at position 225, a leucine (L) at position 227, a threonine (T) at position 231, a lysine (K) at position 241, an isoleucine (I) at position 251, a glycine (G) at

[0498] -95-

[0499] IPTS / 200163498.2 DOCKET NO. SES-020WO PATENT position 261, an alanine (A) at position 273, a glutamic acid (E) at position 274, a glutamic acid (E) at position 278, a proline (P) at position 289, a leucine (L) at position 303, a glutamic acid (E) at position 314, and an isoleucine (I) at position 334, wherein the positions correspond to the numbering as set forth in SEQ ID NO: 35.

[0500] In some embodiments, the polypeptide has an amino acid sequence having at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to the amino acid sequence of SEQ ID NO: 118, provided that the polypeptide comprises a threonine (T) at position 60, an aspartic acid (D) at position 61, an isoleucine (I) at position 63, a tyrosine (Y) at position 64, a leucine (L) at position 69, a lysine (K) at position 74, a glycine (G) at position 76, a leucine (L) at position 85, a glutamic acid (E) at position 112, an asparagine (N) at position 116, a lysine (K) at position 119, a lysine (K) at position 120, a tyrosine (Y) at position 121, a glycine (G) at position 130, a leucine (L) at position 134, an aspartic acid (D) at position 145, a serine (S) at position 146, a threonine (T) at position 148, an aspartic acid (D) at position 154, an aspartic acid (D) at position 163, an aspartic acid (D) at position 190, an arginine (R) at position 198, an asparagine (N) at position 223, a tyrosine (Y) at position 225, a leucine (L) at position 227, a threonine (T) at position 231, a lysine (K) at position 241, an isoleucine (I) at position 251, a glycine (G) at position 261, an alanine (A) at position 273, a glutamic acid (E) at position 274, a glutamic acid (E) at position 278, a proline (P) at position 289, a leucine (L) at position 303, a glutamic acid (E) at position 314, and an isoleucine (I) at position 334, wherein the positions correspond to the numbering as set forth in SEQ ID NO: 35.

[0501] In some embodiments, a polypeptide comprises an amino acid sequence selected from SEQ ID NO: 114, 115, 116, 117, 118, 203, 204, 205, 206, 207, 208, 209, 210, 211, 212, 213, 214, 215, 216, or 217. In some embodiments, a polypeptide comprises an amino acid sequence of SEQ ID NO: 114. In some embodiments, a polypeptide comprises an amino acid sequence of SEQ ID NO: 115. In some embodiments, a polypeptide comprises an amino acid sequence of

[0502] SEQ ID NO: 116. In some embodiments, a polypeptide comprises an amino acid sequence of

[0503] SEQ ID NO: 117. In some embodiments, a polypeptide comprises an amino acid sequence of

[0504] SEQ ID NO: 118. In some embodiments, a polypeptide comprises an amino acid sequence of

[0505] SEQ ID NO: 203. In some embodiments, a polypeptide comprises an amino acid sequence of

[0506] -96-

[0507] IPTS / 200163498.2 DOCKET NO. SES-020WO PATENT

[0508] SEQ ID NO: 204. In some embodiments, a polypeptide comprises an amino acid sequence of SEQ ID NO: 205. In some embodiments, a polypeptide comprises an amino acid sequence of SEQ ID NO: 206. In some embodiments, a polypeptide comprises an amino acid sequence of SEQ ID NO: 207. In some embodiments, a polypeptide comprises an amino acid sequence of SEQ ID NO: 208. In some embodiments, a polypeptide comprises an amino acid sequence of SEQ ID NO: 209. In some embodiments, a polypeptide comprises an amino acid sequence of SEQ ID NO: 210. In some embodiments, a polypeptide comprises an amino acid sequence of SEQ ID NO: 211. In some embodiments, a polypeptide comprises an amino acid sequence of SEQ ID NO: 212. In some embodiments, a polypeptide comprises an amino acid sequence of SEQ ID NO: 213. In some embodiments, a polypeptide comprises an amino acid sequence of SEQ ID NO: 214. In some embodiments, a polypeptide comprises an amino acid sequence of SEQ ID NO: 215. In some embodiments, a polypeptide comprises an amino acid sequence of SEQ ID NO: 216. In some embodiments, a polypeptide comprises an amino acid sequence of SEQ ID NO: 217.

[0509] In some embodiments, the lysine (K) at position 241, wherein the positions correspond to the numbering as set forth in SEQ ID NO: 35, is not mutated. In some embodiments, the lysine (K) at position 241, wherein the positions correspond to the numbering as set forth in SEQ ID NO: 35, is a lysine (K).

[0510] In some embodiments, the polypeptide comprises a polypeptide comprising the sequence of GLSARRI (SEQ ID NO: 122). In some embodiments, the polypeptide comprises a sequence of TDSKLFDYFKEKAFPGLSARRI (SEQ ID NO: 123) or a variant thereof. In some embodiments, the polypeptide comprises a sequence of NLSARQ. In some embodiments, the polypeptide comprises a sequence of TDSKLFDYFKEKAFPNLSARQ (SEQ ID NO: 124), or a variant thereof. The variants of TDSKLFDYFKEKAFPGLSARRI (SEQ ID NO: 123) or TDSKLFDYFKEKAFPNLSARQ (SEQ ID NO: 124) can comprise 1, 2, 3, 4, or 5 substitutions. In some embodiments, variant comprises the peptide with a substitution as provided for herein, provided that a threonine (T) at position 148, an aspartic acid (D) at position 154, an asparagine (N) at position 163, an alanine (A) at position 166, an arginine (R) at position 167, and a glutamine (Q) at position 168, wherein the positions correspond to the numbering as set forth in SEQ ID NO: 35, are not mutated. In some embodiments, variant comprises the peptide with a substitution as provided for herein, provided that a threonine (T) at position 148, an aspartic acid

[0511] -97-

[0512] IPTS / 200163498.2 DOCKET NO. SES-020WO PATENT

[0513] (D) at position 154, an asparagine (N) at position 163, an alanine (A) at position 166, an arginine (R) at position 167, a glutamine (Q) at position 168, and an isoleucine (I) at position 169, wherein the positions correspond to the numbering as set forth in SEQ ID NO: 35, are not mutated.

[0514] As provided for herein, the polypeptides can also comprise a leader sequence attached to the N-terminus of the protein. For example, in some embodiments, the polypeptide has an amino acid sequence having at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to any one amino acid sequence selected from SEQ ID NO: 114, 115, 116, 117, 118, 203, 204, 205, 206, 207, 208, 209, 210, 211, 212, 213, 214, 215, 216, or 217, and further comprises a leader amino acid sequence. In some embodiments, the polypeptide has an amino acid sequence having at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to any one amino acid sequence selected from SEQ ID NO: 114, 115, 116, 117, 118, 203, 204, 205, 206, 207, 208, 209, 210, 211, 212, 213, 214, 215, 216, or 217, and further comprises a leader amino acid sequence having an amino acid sequence of METDTLLLWVLLLWVPGSTG (SEQ ID NO: 109). In some embodiments, the leader amino acid sequence is N-terminal to the polypeptide. In some embodiments, the leader amino acid sequence facilitates expression of the polypeptide. In some embodiments, the leader amino acid sequence is processed and removed from the polypeptide during expression ex vivo or in vivo.

[0515] In some embodiments, the polypeptides or proteases provided for herein do not comprises a protease sequence as illustrated in the following table:

[0516] -98-

[0517] IPTS / 200163498.2 DOCKET NO. SES-020WO PATENT

[0518] -99-

[0519] IPTS / 200163498.2 DOCKET NO. SES-020WO PATENT

[0520] -100-

[0521] IPTS / 200163498.2 DOCKET NO. SES-020WO PATENT

[0522] -101-

[0523] IPTS / 200163498.2 DOCKET NO. SES-020WO PATENT

[0524] -102-

[0525] IPTS / 200163498.2 DOCKET NO. SES-020WO PATENT

[0526] -103-

[0527] IPTS / 200163498.2 DOCKET NO. SES-020WO PATENT

[0528] Thus, in some embodiments, the protease, molecule, or polypeptide does not comprise a protease sequence as provided for in PROV-1, PROV-2, PROV-3, PROV-4, PROV-5, PROV-6,

[0529] PROV-7, PROV-8, PROV-9, PROV-IO, PROV-11, PROV-12, PROV-13, PROV-14, PROV-15,

[0530] PROV-16, PROV-17, PROV-18, PROV-19, PROV-20, PROV-21, PROV-22, PROV-23, PROV-24, PROV-25, PROV-26, PROV-27, PROV-28, PROV-29, PROV-30, PROV-31, PROV-32, PROV-33, PROV-34, PROV-35, PROV-36, PROV-37, PROV-38, PROV-39, PROV-40, PROV-41, PROV-42, PROV-43, PROV-44, PROV-45, PROV-46, PROV-47, PROV-48, PROV-49, PROV-50, PROV-51, PROV-52, PROV-53, PROV-54, PROV-55, PROV-56, PROV-57, PROV-58, or PROV-59. In some embodiments, the protease, molecule, or polypeptide does not comprise an amino acid sequence of any one of SEQ ID NO: 15, 16, 17, 18, 27, 30, 32, 33, 34, 53, 54, 56, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, or 106. In some embodiments, the protease, molecule, or polypeptide does not comprise a protease polypeptide amino acid sequence (domain) as provided for in U.S. Application No. 18 / 405,861, fded January 5, 2024, which is hereby incorporated by reference in its entirety.

[0531] -104-

[0532] IPTS / 200163498.2 DOCKET NO. SES-020WO PATENT

[0533] In some embodiments, a polypeptide comprises an amino acid sequence having at least 50%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%, or 100% sequence identity, or is identical, to any one sequence of VRT-1, VRT-2, VRT-3, or VRT-4.

[0534] In some embodiments, the polypeptide that is less than 100% identical to the reference sequence retains the protease activity and the Fc polypeptide is protease resistant. In some embodiments, the protease resistant Fc polypeptide comprise a Y296Q and / or a P329K mutation. In some embodiments, the protease resistant Fc polypeptide comprises a L234A, L235A, and / or G237A mutation. In some embodiments, the protease resistant Fc polypeptide comprises one, two, or each of L234A, L235A, and / or G237A mutations and one or both of Y296Q and / or a P329K mutations. In some embodiments, the protease resistant Fc polypeptide comprises a set of L234A, L235A, G237A, Y296Q and P329K mutations.

[0535] In some embodiments, a polypeptide comprises an amino acid sequence of any one of VRT-1, VRT-2, VRT-3, or VRT-4.

[0536] In some embodiments, two (or more) polypeptides associate, either covalently or non- covalently, e.g., to form a hetero or homo-dimeric therapeutic compound. In some embodiments, the linker can comprise an Fc region and two Fc regions associate with one another. In some embodiments of a therapeutic compound comprising two linker regions, the linker regions can self-associate, e g., as two identical Fc regions. In some embodiments of a therapeutic compound comprising two linker regions, the linker regions are not capable of, or not capable of substantial, self-association, e.g., the two Fc regions can be members of a knob and hole pair.

[0537] In some embodiments, a polypeptide can associate with another polypeptide. In some embodiments, the polypeptide associated with another polypeptide forms a dimer molecule. In some embodiments, the polypeptide comprises a first polypeptide and a second polypeptide. In some embodiments, the first polypeptide comprises a knob mutation, and the second polypeptide comprises a hole mutation. In some embodiments, the first polypeptide comprises a hole mutation, and the second polypeptide comprises a knob mutation. In some embodiments, the knob mutation is such as those provided herein. In some embodiments, the hole mutation is such as those provided herein. In some embodiments, the knob mutation is any knob mutation known in the art. In some embodiments, the hole mutation is any hole mutation known in the art.

[0538] -105-

[0539] IPTS / 200163498.2 DOCKET NO. SES-020WO PATENT

[0540] In some embodiments, non-limiting examples of knob mutations include T366W, T366Y, Y407R, or S354C mutations as compared to SEQ ID NO: 1, according to EU numbering.

[0541] In some embodiments, non-limiting examples of hole mutations include Y349C, E357T, Y407A, Y407D, or Y407S mutations as compared to SEQ ID NO: 1, according to EU numbering.

[0542] In some embodiments, the Fc region comprises a set of at least one knob mutation selected from T366W, T366Y, Y407R, S354C mutations, or any know mutation known in the art, as compared to SEQ ID NO: 1, according to EU numbering, and at least one hole mutation selected from Y349C, E357T, Y407A, Y407D, Y407S mutations, or any hole mutation known in the art, as compared to SEQ ID NO: 1, according to EU numbering.

[0543] In some embodiments, the knob mutation is such as those provided in SEQ ID NO: 24, or

[0544] DKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMI SRTPEVTCVWDVSHEDPEVKFNWYVDGVEVHNAKTKP REEQQNSTYRWSVLTVLHQDWLNGKEYKCKVSNKALKAPIEKTI SKAKGQPREPQVYTLPPCRDELTKNQ VSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALH NHYTQKSLSLSPG ( SEQ ID NO : 218 ) .

[0545] In some embodiments, the hole mutation is such as those provided in SEQ ID NO: 25, or DKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMI SRTPEVTCVWDVSHEDPEVKFNWYVDGVEVHNAKTKP REEQQNSTYRWSVLTVLHQDWLNGKEYKCKVSNKALKAPIEKTI SKAKGQPREPQVCTLPPSRDELTKNQ VSLSCAVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSKLTVDKSRWQQGNVFSCSVMHEALH NHYTQKSLSLSPG ( SEQ ID NO : 219 ) .

[0546] In some embodiments, a first Fc polypeptide comprises a T366W substitution and the second Fc polypeptide comprises T366S, L368A, and Y407V substitutions. The numbering referenced here in Fc domains is according to EU numbering, which is a standard used for numbering / identifying residues in Fc domains. In some embodiments, the first and second polypeptides that form a heterodimer also comprise cysteine substitutions (a residue on each is changed to a cysteine). In some embodiments, the substitution is at position 354 (EU numbering) and / or position 349 (EU numbering). In some embodiments, the substitution is a S354C substitution. In some embodiments, the substitution is Y349C substitution. Without being bound to any theory, the presence of cysteines on the first and second polypeptides can facilitate the formation of the dimer (homodimer or heterodimer by allowing the formation of disulfide bond. Thus, in some embodiments, the first polypeptide comprises S354C and T366W substitutions and the second polypeptide comprises Y349C, T366S, L368A, and Y407V substitutions. Other

[0547] -106-

[0548] IPTS / 200163498.2 DOCKET NO. SES-020WO PATENT knob-in-hole Fc heterodimer mutations can be used and the cysteine substitutions can also be made in combination with those known knob-in-hole formats.

[0549] Thus, in some embodiments, a dimer molecule comprises a first polypeptide and a second polypeptide, which can be the same or different. If different, the dimer can be referred to as a heterodimer. If the Fc domains are the same, the dimer can be referred to as a heterodimer. In some embodiments, the first molecule comprises a protease conjugated to an Fc region, or an Fc region. In some embodiments, the second molecule comprises a protease conjugated to an Fc region, or an Fc region. In some embodiments, the first polypeptide comprises a protease conjugated to an Fc region, and the second polypeptide comprises an Fc region. Thus, in some embodiments, the second polypeptide comprising an Fc does not contain or promise a protease, such as those provided for herein. A molecule that has two Fc polypeptides and only one protease polypeptide linked to one of the Fc molecules is considered to be a monovalent protease molecule because it only has one protease domain or polypeptide. Thus, in some embodiments, a monovalent protease polypeptide is provided for herein, wherein the polypeptide comprises two Fc domains, which can be as provided for herein, and one protease polypeptide domain linked to one of the Fc domains. In some embodiments, the first polypeptide comprises an Fc region, and the second polypeptide comprises a protease conjugated to an Fc region. In some embodiments, the Fc region comprises knob in hole mutations. In some embodiments, the Fc region forms a dimer. In some embodiments, the Fc region of the first polypeptide and the second polypeptide form a dimer. A protease molecule that is monovalent is illustrated in a non-limiting embodiment in FIG. 1.

[0550] In reference to FIG. 1, the left panel illustrates a non-limiting example of a bivalent protease, which is formed by having a Fc homodimer, where each member of the homodimer is a Fc polypeptide (10). In the homodimer the Fc polypeptides are identical, which is illustrated by them both being labeled as Fci (the subscript for an illustrated Fc domain, whether it is Fci or FC2, is for notation purposes only to show that the Fc domains form a homodimer or heterodimer and is not meant to make a reference to different immunoglobulin subclasses or allotypes). The Ig Protease (20) is linked to the Fc domain, which can be direct, or as illustrated above and provided for herein, via a peptide linker (25). The left panel illustrates the bivalent protease having two Ig Protease domains (20). As used herein, the term “bivalent protease” refers to a polypeptide molecule having two protease polypeptide domains. In the right panel, a non-

[0551] -107-

[0552] IPTS / 200163498.2 DOCKET NO. SES-020WO PATENT limiting example of a monovalent protease polypeptide is illustrated. As illustrated in the right panel, the monovalent protease polypeptide comprises a heterodimer Fc polypeptide, where the heterodimer is made up of a first Fc domain, Fci (10), and a second, different, Fc domain Fc2 (15). The right panel illustrates one Ig protease domain (20) being linked to only one of the Fc polypeptides. As provided for herein, this linkage can be via a peptide linker (25). In some embodiments, the peptide linker (25) is any peptide linker, such as but not limited to those provided herein. Additionally, although as illustrated in FIG. 1, and in certain examples provided for herein, the protease is shown linked to one Fc polypeptide, such as the one containing what are referred to as “knob” substitutions, the protease could also be linked to the other Fc polypeptide, which can be referred to as having “hole” substitutions. Examples of such knob and hole substitutions are provided for herein and are known to one of skill in the art. Thus, the protease polypeptide domain in a monovalent protease can be linked to either of the Fc polypeptides that form the Fc heterodimer.

[0553] Although illustrated as being linked to the C-terminus of the Fc domain in FIG. 1 and in the table below, the Ig protease polypeptide could be linked to the N-terminus of the Fc domain directly or also through a peptide linker as provided for herein.

[0554] In some embodiments, a first polypeptide comprises an amino acid sequence having at least 50%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%, or 100% sequence identity to any one sequence of SEQ ID NO: 57, 58, 59, 60, 189, 190, 191, 192, 193, 194, 195, 196, 197, 198, or 199, wherein said amino acid sequence is conjugated to an Fc region comprising knob in hole mutations, such as those provided herein, for example, and without limitation, SEQ ID NO: 24, 25, 218, and / or 219. In some embodiments, a second polypeptide comprises an Fc region comprising knob in hole mutations, such as those provided herein, for example, and without limitation, SEQ ID NO: 24, 25, 218, and / or 219.

[0555] In some embodiments, a first polypeptide comprises an Fc region comprising knob in hole mutations, such as those provided herein, for example, and without limitation, SEQ ID NO: 24, 25, 218, and / or 219. In some embodiments, a second polypeptide comprises an amino acid sequence having at least 50%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%, or 100% sequence identity to any one sequence of SEQ ID NO: 57,

[0556] -108-

[0557] IPTS / 200163498.2 DOCKET NO. SES-020WO PATENT

[0558] 58, 59, 60, 189, 190, 191, 192, 193, 194, 195, 196, 197, 198, or 199, wherein said amino acid sequence is conjugated to an Fc region comprising knob in hole mutations, such as those provided herein, for example, and without limitation, SEQ ID NO: 24, 25, 218, and / or 219.

[0559] In some embodiments, a dimer molecule comprises a first polypeptide and a second polypeptide. In some embodiments, a first polypeptide comprises an amino acid sequence having at least 50%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%, or 100% sequence identity to any one sequence of SEQ ID NO: 114, 115, 116, 117, 118, 203, 204, 205, 206, 207, 208, 209, 210, 211, 212, 213, 214, 215, 216, or 217. In some embodiments, a second polypeptide comprises an amino acid sequence having at least 50%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%, or 100% sequence identity to any one sequence of SEQ ID NO: 114, 115, 116, 117, 118, 203, 204,

[0560] 205, 206, 207, 208, 209, 210, 211, 212, 213, 214, 215, 216, or 217. In some embodiments, a dimer molecule comprises a first polypeptide comprising an amino acid sequence having at least 50%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%, or 100% sequence identity to any one sequence of SEQ ID NO: 114, 115, 116, 117, or 118; and a second polypeptide comprising an amino acid sequence having at least 50%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%, or 100% sequence identity to any one sequence of SEQ ID NO: 114, 115, 116, 117, 118, 203, 204, 205,

[0561] 206, 207, 208, 209, 210, 211, 212, 213, 214, 215, 216, or 217.

[0562] In some embodiments, a dimer molecule comprises a first polypeptide and a second polypeptide. In some embodiments, a first polypeptide comprises an amino acid sequence of any one sequence of SEQ ID NO: 114, 115, 116, 117, 118, 203, 204, 205, 206, 207, 208, 209, 210, 211, 212, 213, 214, 215, 216, or 217. In some embodiments, a second polypeptide comprises an amino acid sequence of any one sequence of SEQ ID NO: 114, 115, 116, 117, or 118. In some embodiments, a dimer molecule comprises a first polypeptide comprising an amino acid sequence of any one sequence of SEQ ID NO: 114, 115, 116, 117, or 118; and a second

[0563] -109-

[0564] IPTS / 200163498.2 DOCKET NO. SES-020WO PATENT polypeptide comprising an amino acid sequence of any one sequence of SEQ ID NO: 114, 115, 116, 117, 118, 203, 204, 205, 206, 207, 208, 209, 210, 211, 212, 213, 214, 215, 216, or 217.

[0565] In some embodiments, a dimer molecule comprises a first polypeptide and a second polypeptide, wherein the first polypeptide comprises an amino acid sequence having at least 50%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%, or 100% sequence identity to any one sequence of SEQ ID NO: 114, 115, 116, 117, 118, 203, 204, 205, 206, 207, 208, 209, 210, 211, 212, 213, 214, 215, 216, or 217; and the second polypeptide comprises an amino acid sequence of any one of SEQ ID NO: 24, 25, 218, and / or 219. In some embodiments, a dimer molecule comprises a first polypeptide and a second polypeptide, wherein the first polypeptide comprises an amino acid sequence having at least 50%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%, or 100% sequence identity to any one sequence of SEQ ID NO: 118; and the second polypeptide comprises an amino acid sequence of any one of SEQ ID NO: 25. In some embodiments, a dimer molecule comprises a first polypeptide and a second polypeptide, wherein the first polypeptide comprises an amino acid sequence of SEQ ID NO: 118; and the second polypeptide comprises an amino acid sequence of any one of SEQ ID NO: 25.

[0566] In some embodiments, a dimer molecule comprises a first polypeptide and a second polypeptide. In some embodiments, a first polypeptide comprises an amino acid sequence having at least 50%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%, or 100% sequence identity to any one sequence of VRT-1, VRT-2, VRT-3, VRT-4, VRT-5, VRT-6, VRT-7, VRT-8, VRT-9, VRT-10, VRT-11, VRT-12, VRT-13, VRT-14, VRT- 15, VRT-16, VRT-17, VRT-18, VRT-19, or VRT-20. In some embodiments, a second polypeptide comprises an amino acid sequence having at least 50%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%, or 100% sequence identity to any one sequence of VRT-1, VRT-2, VRT-3, VRT-4, VRT-5, VRT-6, VRT-7, VRT-8, VRT-9, VRT-10, VRT-11, VRT-12, VRT-13, VRT-14, VRT-15, VRT-16, VRT-17, VRT-18, VRT-19, or VRT-20. In some embodiments, a dimer molecule comprises a first polypeptide comprising an

[0567] -110-

[0568] IPTS / 200163498.2 DOCKET NO. SES-020WO PATENT amino acid sequence having at least 50%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%, or 100% sequence identity to any one sequence of VRT- 1, VRT-2, VRT-3, VRT-4, VRT-5, VRT-6, VRT-7, VRT-8, VRT-9, VRT-10, VRT-11, VRT-12, VRT-13, VRT-14, VRT-15, VRT-16, VRT-17, VRT-18, VRT-19, or VRT-20; and a second polypeptide comprising an amino acid sequence having at least 50%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%, or 100% sequence identity to any one sequence of VRT-1, VRT-2, VRT-3, VRT-4, VRT-5, VRT-6, VRT-7, VRT-8, VRT-9, VRT-10, VRT-11, VRT-12, VRT-13, VRT-14, VRT-15, VRT-16, VRT-17, VRT-18, VRT-19, or VRT-20.

[0569] In some embodiments, a dimer molecule comprises a first polypeptide and a second polypeptide. In some embodiments, a first polypeptide comprises an amino acid sequence of any one sequence of VRT-1, VRT-2, VRT-3, VRT-4, VRT-5, VRT-6, VRT-7, VRT-8, VRT-9, VRT-10, VRT-11, VRT-12, VRT-13, VRT-14, VRT-15, VRT-16, VRT-17, VRT-18, VRT-19, or VRT-20.

[0570] In some embodiments, a second polypeptide comprises an amino acid sequence of any one sequence of VRT-1, VRT-2, VRT-3, VRT-4, VRT-5, VRT-6, VRT-7, VRT-8, VRT-9, VRT-10, VRT-11, VRT-12, VRT-13, VRT-14, VRT-15, VRT-16, VRT-17, VRT-18, VRT-19, or VRT-20.

[0571] In some embodiments, a dimer molecule comprises a first polypeptide comprising an amino acid sequence of any one sequence of VRT-1, VRT-2, VRT-3, VRT-4, VRT-5, VRT-6, VRT-7, VRT-8, VRT-9, VRT-10, VRT-11, VRT-12, VRT-13, VRT-14, VRT-15, VRT-16, VRT-17, VRT-18, VRT-19, or VRT-20; and a second polypeptide comprising an amino acid sequence of any one sequence of VRT-1, VRT-2, VRT-3, VRT-4, VRT-5, VRT-6, VRT-7, VRT-8, VRT-9, VRT-10, VRT-11, VRT-12, VRT-13, VRT-14, VRT-15, VRT-16, VRT-17, VRT-18, VRT-19, or VRT-20.

[0572] In some embodiments, the dimer is a homodimer molecule. In some embodiments, the dimer is a heterodimer molecule.

[0573] As used herein, the term “non-covalently conjugated” can mean that a polypeptide is tethered to another polypeptide through a linker. In some embodiments, the linker is a peptide

[0574] -111-

[0575] IPTS / 200163498.2 DOCKET NO. SES-020WO PATENT linker. Non-limiting examples of peptide linkers that can be used are known in the art and are provide for herein.

[0576] As discussed herein the different domains, molecules, or polypeptide can be linked together with a linker domain or region. Any linker region described herein can be used as a linker. Linkers can be for example, gly cine / serine linkers. In some embodiments, the linker can comprise one or more repeats of GGGGS (SEQ ID NO: 174). In some embodiments, the linker comprises 1, 2, 3, 4, or 5 repeats. In some embodiments, the linker comprises the sequence of GGGGSGGGGS (SEQ ID NO: 175). In some embodiments, the linker comprises the sequence of GGGGSGGGGSGGGGS (SEQ ID NO: 176). In some embodiments, the linker comprises the sequence of GGGGSGGGGSGGGGSG (SEQ ID NO: 169). In some embodiments, the linker comprises the amino acid sequence of GGGSK (SEQ ID NO: 172, which can be repeated 1-4 times, which can be represented as (GGGSK)n, where n is 1-4 (SEQ ID NO: 172, SEQ ID NO: 121, SEQ ID NO: 173, SEQ ID NO: 186, respectively). Thus, in some embodiments, the linker can comprise the amino acid sequence of GGGSKGGGSK (SEQ ID NO: 121), GGGSKGGGSKGGGSK (SEQ ID NO: 173), or GGGSKGGGSKGGGSKGGGSK (SEQ ID NO: 186). In some embodiments, the linker comprises the sequence of GGGGSGGGGSGGGGSGGGGSG (SEQ ID NO: 187). In some embodiments, the linker comprises the sequence of GGGGSGGGGSG (SEQ ID NO: 188). In some embodiments, the linker comprises: GGGGS (SEQ ID NO: 174), (GGGGS)3(SEQ ID NO: 176), (GGGGS)n (n=l, 2, 3, 4) (SEQ ID NO: 174, SEQ ID NO: 175, SEQ ID NO: 176, SEQ ID NO: 177, respectively) , (Gly)s (SEQ ID NO: 178), (Gly)6(SEQ ID NO: 179), (EAAAK)3(SEQ ID NO: 180), (EAAK)n (n=l-3) (SEQ ID NO: 181, SEQ ID NO: 182, SEQ ID NO: 183, respectively), A(EAAAK)4ALEA(EAAAK)4A (SEQ ID NO: 184), or AEAAAKEAAAKA (SEQ ID NO: 185). These linkers can be used in any of the compounds or compositions provided herein.

[0577] Antibody molecule, as that term is used herein, refers to a polypeptide, e.g., an immunoglobulin chain or fragment thereof, comprising at least one functional immunoglobulin variable domain sequence. An antibody molecule encompasses antibodies (e.g., full-length antibodies) and antibody fragments. In some embodiments, an antibody molecule comprises an antigen binding or functional fragment of a full-length antibody, or a full length immunoglobulin chain. For example, a full-length antibody is an immunoglobulin (Ig) molecule (e.g., an IgG antibody) that is naturally occurring or formed by normal immunoglobulin gene fragment

[0578] -112-

[0579] IPTS / 200163498.2 DOCKET NO. SES-020WO PATENT recombinatorial processes). Tn embodiments, an antibody molecule refers to an immunologically active, antigen-binding portion of an immunoglobulin molecule, such as an antibody fragment. An antibody fragment, e.g., functional fragment, comprises a portion of an antibody, e.g., Fab, Fab1, F(ab')2, F(ab)2, variable fragment (Fv), domain antibody (dAb), or single chain variable fragment (scFv). A functional antibody fragment binds to the same antigen as that recognized by the intact (e.g., full-length) antibody. The terms “antibody fragment” or “functional fragment” also include isolated fragments consisting of the variable regions, such as the “Fv” fragments consisting of the variable regions of the heavy and light chains or recombinant single chain polypeptide molecules in which light and heavy variable regions are connected by a peptide linker (“scFv proteins”). In some embodiments, an antibody fragment does not include portions of antibodies without antigen binding activity, such as Fc fragments or single amino acid residues. Exemplary antibody molecules include full length antibodies and antibody fragments, e.g., dAb (domain antibody), single chain, Fab, Fab’, and F(ab’)2 fragments, and single chain variable fragments (scFvs).

[0580] The term “antibody molecule” also encompasses whole or antigen binding fragments of domain, or single domain, antibodies, which can also be referred to as “sdAb” or “VHH.” Domain antibodies comprise either VH or VL that can act as stand-alone, antibody fragments. Additionally, domain antibodies include heavy-chain-only antibodies (HCAbs). Domain antibodies also include a CH2 domain of an IgG as the base scaffold into which CDR loops are grafted. It can also be generally defined as a polypeptide or protein comprising an amino acid sequence that is comprised of four framework regions interrupted by three complementarity determining regions. This is represented as FP1- CDP1 -FR2-CDR2-FN1-CDN1-FR4. sdAbs can be produced in camelids such as llamas, but can also be synthetically generated using techniques that are well known in the art. The numbering of the amino acid residues of a sdAb or polypeptide is according to the general numbering for VH domains given by Kabat et al. ("Sequence of proteins of immunological interest," US Public Health Services, NIH Bethesda, MD, Publication No. 91, which is hereby incorporated by reference). According to this numbering, FP1 of a sdAb comprises the amino acid residues at positions 1-30, CDP1 of a sdAb comprises the amino acid residues at positions 31-36, FR2 of a sdAb comprises the amino acids at positions 36-49, CDR2 of a sdAb comprises the amino acid residues at positions 50-65, FN1 of a sdAb comprises the amino acid residues at positions 66- 94, CDN1 of a sdAb comprises the

[0581] -113-

[0582] IPTS / 200163498.2 DOCKET NO. SES-020WO PATENT amino acid residues at positions 95-102, and FR4 of a sdAb comprises the amino acid residues at positions 103-113. Domain antibodies are also described in W02004041862 and WO2016065323, each of which is hereby incorporated by reference. The domain antibodies can be a targeting moiety as described herein.

[0583] Antibody molecules can be monospecific (e.g., monovalent or bivalent), bispecific (e.g., bivalent, trivalent, tetravalent, pentavalent, or hexavalent), trispecific (e.g., trivalent, tetravalent, pentavalent, hexavalent), or with higher orders of specificity (e.g, tetraspecific) and / or higher orders of valency beyond hexavalency. An antibody molecule can comprise a functional fragment of a light chain variable region and a functional fragment of a heavy chain variable region, or heavy and light chains may be fused together into a single polypeptide.

[0584] Effector, as that term is used herein, refers to an entity, e.g., a cell or molecule, e.g., a soluble or cell surface molecule, which mediates an immune response. In some embodiments, the effector is an antibody. In some embodiments, the effectors binding domains as provided for herein, refers to a polypeptide (e.g.) that has sufficient binding specificity that it can bind the effector with sufficient specificity that it can serve as an effector binding / modulating molecule. In some embodiments, it binds to effector with at least 10, 20, 30, 40, 50, 60, 70, 80, 90, or 95% of the affinity of the naturally occurring counter-ligand. In some embodiments, it has at least 60, 70, 80, 90, 95, 99, or 100% sequence identity ,or substantial sequence identity, with a naturally occurring counter-ligand for the effector.

[0585] Elevated risk, as used herein, refers to the risk of a disorder in a subject, wherein the subject has one or more of a medical history of the disorder or a symptom of the disorder, a biomarker associated with the disorder or a symptom of the disorder, or a family history of the disorder or a symptom of the disorder.

[0586] The domains can have similarity to those as provided for herein or those that are incorporated by reference. Sequence identity, percentage identity, and related terms, as those terms are used herein, refer to the relatedness of two sequences, e.g., two nucleic acid sequences or two amino acid or polypeptide sequences. In the context of an amino acid sequence, the term "substantially identical" is used herein to refer to a first amino acid that contains a sufficient or minimum number of amino acid residues that are i) identical to, or ii) conservative substitutions of aligned amino acid residues in a second amino acid sequence such that the first and second amino acid sequences can have a common structural domain and / or common functional activity.

[0587] -114-

[0588] IPTS / 200163498.2 DOCKET NO. SES-020WO PATENT

[0589] For example, amino acid sequences that contain a common structural domain having at least about 85%, 90%. 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to a reference sequence, e.g., an amino acid sequence provided herein.

[0590] In the context of nucleotide sequence, such as those encoding for the domains, the term "substantially identical" is used herein to refer to a first nucleic acid sequence that contains a sufficient or minimum number of nucleotides that are identical to aligned nucleotides in a second nucleic acid sequence such that the first and second nucleotide sequences encode a polypeptide having common functional activity, or encode a common structural polypeptide domain or a common functional polypeptide activity. For example, nucleotide sequences having at least about 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to a reference sequence, e.g., an amino acid sequence provided herein.

[0591] The term “functional variant” refers to polypeptides that have a substantially identical amino acid sequence to the naturally-occurring sequence, or are encoded by a substantially identical nucleotide sequence, and are capable of having one or more activities of the naturally- occurring sequence. For example, a Fc variant can have the amino acid sequence of a Fc polypeptide but comprise a mutation that prevents or disrupts cleavage by a protease.

[0592] Calculations of homology or sequence identity between sequences (the terms are used interchangeably herein) can be performed as follows.

[0593] To determine the percent identity of two amino acid sequences, or of two nucleic acid sequences, the amino acid sequences are aligned for optimal comparison purposes (e.g., gaps can be introduced in one or both of a first and a second amino acid or nucleic acid sequence for optimal alignment and non-homologous sequences can be disregarded for comparison purposes). In a preferred embodiment, the length of a reference sequence aligned for comparison purposes is at least 30%, preferably at least 40%, more preferably at least 50%, 60%, and even more preferably at least 70%, 80%, 90%, 100% of the length of the reference sequence. The amino acid residues or nucleotides at corresponding amino acid positions or nucleotide positions are then compared. When a position in the first sequence is occupied by the same amino acid residue or nucleotide as the corresponding position in the second sequence, then the molecules are identical at that position (as used herein amino acid or nucleic acid "identity" is equivalent to amino acid or nucleic acid "homology").

[0594] -115-

[0595] IPTS / 200163498.2 DOCKET NO. SES-020WO PATENT

[0596] The percent identity between the two sequences is a function of the number of identical positions shared by the amino acid sequences, taking into account the number of gaps, and the length of each gap, which need to be introduced for optimal alignment of the two sequences.

[0597] The comparison of sequences and determination of percent identity between two sequences can be accomplished using a mathematical algorithm. In some embodiments, the percent identity between two amino acid sequences is determined using the Needleman and Wunsch ((1970) J. Mol. Biol. 48:444-453 ) algorithm which has been incorporated into the GAP program in the GCG software package (available at http: / / www.gcg.com), using either a Blossum 62 matrix or a PAM250 matrix, and a gap weight of 16, 14, 12, 10, 8, 6, or 4 and a length weight of 1, 2, 3, 4, 5, or 6. In some embodiments, the percent identity between two nucleotide sequences is determined using the GAP program in the GCG software package (available at http: / / www.gcg.com), using a NWSgapdna.CMP matrix and a gap weight of 40, 50, 60, 70, or 80 and a length weight of 1, 2, 3, 4, 5, or 6. A particularly preferred set of parameters (and the one that should be used unless otherwise specified) are a Blossum 62 scoring matrix with a gap penalty of 12, a gap extend penalty of 4, and a frameshift gap penalty of 5.

[0598] The percent identity between two amino acid or nucleotide sequences can be determined using the algorithm of E. Meyers and W. Miller ((1989) CABIOS, 4: 11-17) which has been incorporated into the ALIGN program (version 2.0), using a PAM120 weight residue table, a gap length penalty of 12 and a gap penalty of 4.

[0599] The nucleic acid and protein sequences described herein can be used as a "query sequence" to perform a search against public databases to, for example, identify other family members or related sequences. Such searches can be performed using the NBLAST and XBLAST programs (version 2.0) of Altschul, et al. (1990) J. Mol. Biol. 215:403-10. BLAST nucleotide searches can be performed with the NBLAST program, score = 100, wordlength = 12 to obtain nucleotide sequences homologous to for example any a nucleic acid sequence provided herein. BLAST protein searches can be performed with the XBLAST program, score = 50, wordlength = 3 to obtain amino acid sequences homologous to protein molecules provided herein. To obtain gapped alignments for comparison purposes, Gapped BLAST can be utilized as described in Altschul et al., (1997) Nucleic Acids Res. 25:3389-3402. When utilizing BLAST and Gapped BLAST programs, the default parameters of the respective programs (e.g., XBLAST and NBLAST) can be used. See http: / / www.ncbi.nlm.nih.gov.

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[0601] IPTS / 200163498.2 DOCKET NO. SES-020WO PATENT

[0602] As used herein, the term “hybridizes under low stringency, medium stringency, high stringency, or very high stringency conditions” describes conditions for hybridization and washing. Guidance for performing hybridization reactions can be found in Current Protocols in Molecular Biology, John Wiley & Sons, N.Y. (1989), 6.3.1-6.3.6, which is incorporated by reference. Aqueous and nonaqueous methods are described in that reference and either can be used. Specific hybridization conditions referred to herein are as follows: 1) low stringency hybridization conditions in 6X sodium chloride / sodium citrate (SSC) at about 45°C, followed by two washes in 0.2X SSC, 0.1% SDS at least at 50°C (the temperature of the washes can be increased to 55°C for low stringency conditions); 2) medium stringency hybridization conditions in 6X SSC at about 45°C, followed by one or more washes in 0.2X SSC, 0.1% SDS at 60°C; 3) high stringency hybridization conditions in 6X SSC at about 45°C, followed by one or more washes in 0.2X SSC, 0.1% SDS at 65°C; and preferably 4) very high stringency hybridization conditions are 0.5M sodium phosphate, 7% SDS at 65°C, followed by one or more washes at 0.2X SSC, 1% SDS at 65°C. Very high stringency conditions (4) are the preferred conditions and the ones that should be used unless otherwise specified.

[0603] It is understood that the molecules of the present embodiments may have additional conservative or non-essential amino acid substitutions, which do not have a substantial effect on their functions.

[0604] The term "amino acid" is intended to embrace all molecules, whether natural or synthetic, which include both an amino functionality and an acid functionality and capable of being included in a polymer of naturally-occurring amino acids. Exemplary amino acids include naturally-occurring amino acids; analogs, derivatives and congeners thereof; amino acid analogs having variant side chains; and all stereoisomers of any of any of the foregoing. As used herein the term "amino acid" includes both the D- or L- optical isomers and peptidomimetics.

[0605] A "conservative amino acid substitution" is one in which the amino acid residue is replaced with an amino acid residue having a similar side chain. Families of amino acid residues having similar side chains have been defined in the art. These families include amino acids with basic side chains (e.g., lysine, arginine, histidine), acidic side chains (e.g., aspartic acid, glutamic acid), uncharged polar side chains (e.g., glycine, asparagine, glutamine, serine, threonine, tyrosine, cysteine), nonpolar side chains (e.g., alanine, valine, leucine, isoleucine, proline,

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[0607] IPTS / 200163498.2 DOCKET NO. SES-020WO PATENT phenylalanine, methionine, tryptophan), beta-branched side chains (e.g., threonine, valine, isoleucine) and aromatic side chains (e.g., tyrosine, phenylalanine, tryptophan, histidine).

[0608] The molecules and polypeptides provided for herein can be used to treat diseases and conditions mediated by IgG. Thus, embodiments are provided for methods of treating an IgG mediated disease or disorder in a subject. In some embodiments, the IgG mediated disease or disorder is an autoimmune disease or disorder. In some embodiments, the methods comprise administering to the subject a polypeptide, molecule, compound, or compositions as provided for herein. In some embodiments, the subject has or is at risk of having an IgG mediated disease or disorder. In some embodiments, the subject has or is at risk of having an autoimmune disease or disorder.

[0609] Antibody-mediated rejection (AMR) causes severe and rapid dysfunction and loss of allografts. Without wishing to be bound to a particular theory, the most common mechanism underlying AMR is an anamnestic response that originates from previous antigenic exposure. These donor specific antibody (DSA) responses are usually robust and result in the rapid production of high levels of DSA and acute allograft dysfunction. The mechanism of injury in AMR involves antigens that initiate the production of DSAs resulting in antigen-antibody interactions, complement activation and inflammation, and the resultant donor tissue damage. The impact of AMR on graft survival is dramatic and continues long after the initial inflammatory condition has resolved as was recently demonstrated in a study by LeFaucheur and Glotz. In this single center study of a large cohort of sensitized recipients, the investigators compared allograft survival for recipients successfully treated for AMR versus those that never experienced AMR. In some embodiments, the molecules and polypeptides provided for herein can be used to treat AMR.

[0610] In some embodiments, methods of treating a disease or disorder in a subject are provided. In some embodiments, the disease or disorder is an Ig mediated disease or disorder. In some embodiments, the disease or disorder is an autoimmune disease or disorder.

[0611] In some embodiments, methods of decreasing circulating immunoglobulins in a subject are provided. In some embodiments, the circulating immunoglobulins are selected from IgG (such as IgGl, IgG2, IgG3, IgG4), IgA, IgM, IgE, or IgD. In some embodiments, methods of decreasing circulating IgG immunoglobulins are provided. In some embodiments, methods of decreasing circulating IgGl immunoglobulins are provided. In some embodiments, methods of

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[0613] IPTS / 200163498.2 DOCKET NO. SES-020WO PATENT decreasing circulating TgG2 immunoglobulins are provided. In some embodiments, methods of decreasing circulating IgG3 immunoglobulins are provided. In some embodiments, methods of decreasing circulating IgG4 immunoglobulins are provided. In some embodiments, methods of decreasing circulating IgA immunoglobulins are provided. In some embodiments, methods of decreasing circulating IgM immunoglobulins are provided. In some embodiments, methods of decreasing circulating IgE immunoglobulins are provided. In some embodiments, methods of decreasing circulating IgD immunoglobulins are provided. In some embodiments, methods of decreasing circulating immunoglobulins in a subject are provided, wherein the subject is a subject with an autoimmune disease or disorder.

[0614] In some embodiments, a method of reducing immunoglobulins is provided. In some embodiments, a method of reducing immunoglobulins, such as IgG, in a subject, is provided. In some embodiments, the method of reducing immunoglobulins, such as IgG, in a subject, comprises administering to the subject any one of the polypeptides or compositions provided for herein. In some embodiments, the method of reducing immunoglobulins, such as IgG, in a subject, comprises administering to the subject the polypeptide comprising the amino acid sequence of any one of SEQ ID NO: 57, 58, 59, 60, 189, 190, 191, 192, 193, 194, 195, 196, 197, 198, or 199, or the composition thereof. In some embodiments, the reduction of immunoglobulins may be about 5%, about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, or about 100% reduction. In some embodiments, the reduction of immunoglobulins may be at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, or at least 95% reduction. In some embodiments, the reduction of immunoglobulins may be a 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% reduction.

[0615] In some embodiments, a method of treating a disease or disorder in a subject is provided. In some embodiments, the method of treating a disease or disorder in a subject, comprises administering to the subject any one of the polypeptides or compositions provided for herein. In some embodiments, the method of treating a disease or disorder in a subject comprises administering to the subject the polypeptide comprising the amino acid sequence of any one of

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[0617] IPTS / 200163498.2 DOCKET NO. SES-020WO PATENT

[0618] SEQ ID NO: 57, 58, 59, 60, 189, 190, 191, 192, 193, 194, 195, 196, 197, 198, or 199, or the composition thereof.

[0619] In some embodiments, a method of treating a transplant subject is provided. In some embodiments, the method of treating a transplant subject comprises administering to the subject any one of the polypeptides or compositions provided for herein. In some embodiments, the method of treating a transplant subject comprises administering to the subject the polypeptide comprising the amino acid sequence of any one of SEQ ID NO: 57, 58, 59, 60, 189, 190, 191, 192, 193, 194, 195, 196, 197, 198, or 199, or the composition thereof.

[0620] In some embodiments, a method of improving a gene-therapy in subject is provided. In some embodiments, the method of improving a gene-therapy in subject comprises administering to the subject any one of the polypeptides or compositions provided for herein. In some embodiments, the method of improving a gene-therapy in subject comprises administering to the subject the polypeptide comprising the amino acid sequence of any one of SEQ ID NO: 57, 58, 59, 60, 189, 190, 191, 192, 193, 194, 195, 196, 197, 198, or 199, or the composition thereof.

[0621] In some embodiments, a method of treating a subject having, or at risk, or elevated risk, for having, an IgG mediated disease or disorder is provided. In some embodiments, the method of treating a subject having, or at risk, or elevated risk, for having, an IgG mediated disease or disorder comprises administering to the subject any one of the polypeptides or compositions provided for herein. In some embodiments, the method of treating a subject having, or at risk, or elevated risk, for having, an IgG mediated disease or disorder comprises administering to the subject the polypeptide comprising the amino acid sequence of any one of SEQ ID NO: 57, 58, 59, 60, 189, 190, 191, 192, 193, 194, 195, 196, 197, 198, or 199, or the composition thereof.

[0622] In some embodiments, a method of cleaving B cell receptor in a subject is provided. In some embodiments, the method of cleaving B cell receptor in a subject comprises administering to the subject any one of the polypeptides or compositions provided for herein. In some embodiments, the method of cleaving B cell receptor in a subject comprises administering to the subject the polypeptide comprising the amino acid sequence of any one of SEQ ID NO: 57, 58, 59, 60, 189, 190, 191, 192, 193, 194, 195, 196, 197, 198, or 199, or the composition thereof.

[0623] Non-limiting examples of diseases and disorders, such as autoimmune diseases or disorders, that can be treated with the molecules and polypeptides described herein include, but are not limited to, Addison's disease, Anti-GBM glomerulonephritis, anti -neutrophil cytoplasmic

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[0625] IPTS / 200163498.2 DOCKET NO. SES-020WO PATENT antibody-associated vasculitis, ANCA associated vasculitis, granulomatous polyangiitis (Wegener granulomatosis), eosinophilic granulomatosis with polyangiitis (Churg-Strauss syndrome), microscopic polyangiitis, anti-NMDAR encephalitis, anti-phospholipid antibody syndrome (APS) and catastrophic APS, autoimmune bullous skin diseases, pemphigus, pemphigus foliaceus (PF), fogo selvage (FS), pemphigus vulgaris (PV), autoimmune hemolytic anemia (AIHA), autoimmune hepatitis (AIH), autoimmune neutropenia (AIN), bullous pemphigoid (BP), Celiac’s disease, chronic urticaria, complete congenital heart block (CCHB), diabetes type 1A (T1DM), epidermolysis bullosa acquisita (EBA), essential mixed cryoglobulinemia, Goodpasture’s syndrome, anti-glomerular basement membrane disease, Graves’ disease, Goitre, hyperthyroidism, infiltrative exophthalmos, infiltrative dermopathy, Guillain-Barre syndrome (GBS), acute inflammatory demyelinating polyneuropathy (AIDP), acute motor axonal neuropathy (AMAN), hemophilia, acquired FVII deficiency, idiopathic thrombocytopenic purpura (ITP), Lambert-Eaton myasthenic syndrome (LEMS), mixed connective tissue disease (MCTD), multiple myeloma, myasthenia gravis, myasthenic crisis, myocarditis, dilated cardiomyopathy (DCM), congestive cardiomyopathy, neuromyelitis optica (NMO), primary biliary cirrhosis (PBC), primary progressive multiple sclerosis (PPMS), relapsing-rheumatic multiple sclerosis, acute exacerbation of multiple sclerosis, rheumatic heart disease (RHD), rheumatoid arthritis (RA), serum-sickness, immune complex hypersensitivity (type III), Sjogren’s syndrome (SS), lupus nephritis, systemic lupus erythematosus (SLE), cutaneous lupus (CLE), transplant rejection, thrombotic thrombocytopenic purpura (TTP), idiopathic inflammatory myopathies, polymyositis, dermatomyositis, inclusion body myositis, uveitis, scleritis, ankylosing spondylitis, axial spondylarthritis, reactive arthritis, psoriatic arthritis, IBD-associated arthritis, antiphospholipid syndrome, systemic sclerosis, giant cell arteritis, polymyalgia rheumatic, Takayasu’s arteritis, anti-neutrophil cytoplasmic antibody associated vasculitis, Henoch-Schonlein purpura, polyarteritis nodosa, Cogan’s syndrome, Buerger’s disease, Susan’s disease, immune complex vasculitis, primary angiitis of the central nervous system (CNS), Behcet’s disease, relapsing polychondritisjuvenile idiopathic arthritis, juvenile dermatomyositis, autoimmune brain disease, sarcoidosis, neurosarcoidosis, IgG4-related diseases, autoimmune complications of immune checkpoint inhibitors (IRAEs), adult onset Still’s disease, warm antibody hemolytic anemia (wAIHA), immune thrombocytopenia, immune thrombotic thrombocytopenia, thrombic thrombocytopenia, pernicious anemia, aplastic anemia,

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[0627] IPTS / 200163498.2 DOCKET NO. SES-020WO PATENT

[0628] Evan’s syndrome, autoimmune neutropenia, acquired von Willibrand syndrome, recurring fetal loss, Rh mismatch, ulcerative colitis, autoimmune hepatitis, autoimmune pancreatitis, eosinophilic esophagitis / gastritis, primary sclerosing cholangitis, primary biliary sclerosis, glomerulonephritis, glomerular basement membrane disease, scleritis / episcleritis, uveitis, conjunctivitis, keratitis, type I diabetes mellitus, Hashimoto’s thyroiditis, polyglandular autoimmune endocrine syndromes, atopic dermatitis, dermatitis herpetiformis, pemphigus foliaceus, cutaneous lupus erythematosus, linear IgA disease, Lichen planus, transplantation, antibody-mediated rejection, alloantibody hypersensitization, xenoantibody mediated rejection, solid organ rejection, graft vs. host disease (GVHD), multiple sclerosis, neuromyelitis optica, amyotrophic lateral sclerosis, Parkinson’s disease, autoimmune encephalitis, CNS vasculitis, chronic idiopathic demyelinating polyneuropathy, transverse myelitis, optic neuritis, anti-myelin oligodendrocyte glycoprotein (MOG) disease, chronic meningitis, rheumatic heart disease, infectious disease, vaccination, antibody dependent enhancement, antibody to therapeutic biologic agents (cytokines, monoclonal antibodies, enzymes, coagulation factors), allergic asthma, eosinophilic pneumonia, nonspecific interstitial pneumonia, rheumatoid arthritis- associated interstitial lung disease (RA-ILD), sarcoidosis, hypersensitive pneumonitis, allergic bronchopulmonary mycosis, chronic rhinosinusitis with nasal polyps, COVID-19, long COVID- 19, and the like.

[0629] In some embodiments, methods of treating a transplant subject are provided. In some embodiments, methods of improving a gene-therapy in a subject are provided. In some embodiments, methods of treating a subject having, or at risk, or elevated risk, for having, an IgG mediated disease or disorder are provided.

[0630] In some embodiments, methods comprise administering a therapeutically effective amount of any one of the polypeptides provided herein, or the pharmaceutical composition comprising any one polypeptide provided herein, thereby treating the subject.

[0631] In some embodiments, the condition or disease to be treated is a neoplastic disorder, such as a cancer. In some embodiments, the cancer is a solid or liquid tumor. In some embodiments, the liquid or solid tumor include, but are not limited to, hematopoietic cancer, lymphoid cancer, skin cancer, head and neck cancer, genitourinary cancer, blood cancer, lung cancer, breast cancer, brain cancer, esophageal cancer, colorectal cancer, pancreatic cancer, and any

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[0633] IPTS / 200163498.2 DOCKET NO. SES-020WO PATENT combination thereof.

[0634] Therapeutic Antibodies

[0635] The polypeptides provided herein may be co-administered with other therapeutics. In some embodiments, the other therapeutic is a therapeutic antibody. In some embodiments, the therapeutic antibody is an antibody used for treatment of a disease, such as and without limitation, cancer, autoimmune diseases, infectious diseases, inflammatory diseases, cardiovascular diseases, neurological disorders, ophthalmic diseases, and respiratory diseases. In some embodiments, the therapeutic antibody is selected from Table 10.

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[0649] IPTS / 200163498.2 DOCKET NO. SES-020WO PATENT

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[0651] IPTS / 200163498.2 DOCKET NO. SES-020WO PATENT

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[0653] IPTS / 200163498.2 DOCKET NO. SES-020WO PATENT

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[0655] IPTS / 200163498.2 DOCKET NO. SES-020WO PATENT

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[0657] IPTS / 200163498.2 DOCKET NO. SES-020WO PATENT

[0658] The therapeutic antibodies provided for herein are known in the art. Thus, in some embodiments, all elements of the therapeutic antibody, such as and without limitation, heavy chains, light chains, variable regions, constant regions, antigen-binding sites, the Fc region, the hinge region, disulfide bonds, glycosylation sites, and framework regions may be known in the art. In some embodiments, the heavy chain (HC) and light chain (LC) of each therapeutic antibody provided for herein, without limitation in Table 10, is known in the art. As used herein, the therapeutic antibody selected from Rituxan (rituximab), Ocrevus (ocrelizumab), Orencia (abatacept), Stelara (ustekinumab), Cosentyx (secukinumab), Tremfya (guselkumab), Skyrizi (risankizumab), Dupixent (dupilumab), Actemra (tocilizumab), Enbrel (etanercept), Humira (adalimumab), Tysabri (natalizumab), Benlysta (belimumab), Saphnelo (anifrolumab), Xolair (omalizumab), Taltz (ixekizumab), Bimzelx (bimekizumab), Nucala (mepolizumab), Fasenra (benralizumab), Prolia (denosumab), Remicade (infliximab), Simponi (golimumab), Entyvio (vedolizumab), Briumvi (ublituximab), Orthoclone Okt3 (Muromonab-CD3), MabCampath / Lemtrada) (lemtuzumab), Alzumab (itolizumab), Veopoz (pozelimab), Soliris (eculizumab), Siliq (brodalumab), Haris (canakinumab), Arcalyst (rilonacept), Omvoh (mirikizumab), Ilumya (tildrakizumab), Simulect (basiliximab), Zenapax (daclizumab), Spevigo

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[0660] IPTS / 200163498.2 DOCKET NO. SES-020WO PATENT

[0661] (spesolimab), Cinqair (reslizumab), Kevzara (sarilumab), Sylvant (siltuximab), Adbry (tralokinumab), Ebglyss (lebrikizumab), is meant to comprises a heavy chain (HC) and a light chain (LC) each of which is known in the art to be the heavy chain (HC) and the light chain (LC) of said therapeutic antibody. In some embodiments, the therapeutic antibody selected from Rituxan (rituximab), Ocrevus (ocrelizumab), Orencia (abatacept), Stelara (ustekinumab), Cosentyx (secukinumab), Tremfya (guselkumab), Skyrizi (risankizumab), Dupixent (dupilumab), Actemra (tocilizumab), Enbrel (etanercept), Humira (adalimumab), Tysabri (natalizumab), Benlysta (belimumab), Saphnelo (anifrolumab), Xolair (omalizumab), Taltz (ixekizumab), Bimzelx (bimekizumab), Nucala (mepolizumab), Fasenra (benralizumab), Prolia (denosumab), Remicade (infliximab), Simponi (golimumab), Entyvio (vedolizumab), Briumvi (ublituximab), Orthoclone Okt3 (Muromonab-CD3), MabCampath / Lemtrada) (lemtuzumab), Alzumab (itolizumab), Veopoz (pozelimab), Soliris (eculizumab), Siliq (brodalumab), Haris (canakinumab), Arcalyst (rilonacept), Omvoh (mirikizumab), Ilumya (tildrakizumab), Simulect (basiliximab), Zenapax (daclizumab), Spevigo (spesolimab), Cinqair (reslizumab), Kevzara (sarilumab), Sylvant (siltuximab), Adbry (tralokinumab), Ebglyss (lebrikizumab), is meant to comprises a heavy chain (HC) comprising mutations to the Fc domain of the heavy chain, such as those provided for herein, and a light chain (LC) known in the art to be the light chain (LC) of said therapeutic antibody. Accordingly, in some embodiments, the therapeutic antibody selected from Rituxan (rituximab), Ocrevus (ocrelizumab), Orencia (abatacept), Stelara (ustekinumab), Cosentyx (secukinumab), Tremfya (guselkumab), Skyrizi (risankizumab), Dupixent (dupilumab), Actemra (tocilizumab), Enbrel (etanercept), Humira (adalimumab), Tysabri (natalizumab), Benlysta (belimumab), Saphnelo (anifrolumab), Xolair (omalizumab), Taltz (ixekizumab), Bimzelx (bimekizumab), Nucala (mepolizumab), Fasenra (benralizumab), Prolia (denosumab), Remicade (infliximab), Simponi (golimumab), Entyvio (vedolizumab), Briumvi (ublituximab), Orthoclone Okt3 (Muromonab-CD3), MabCampath / Lemtrada) (lemtuzumab), Alzumab (itolizumab), Veopoz (pozelimab), Soliris (eculizumab), Siliq (brodalumab), Haris (canakinumab), Arcalyst (rilonacept), Omvoh (mirikizumab), Ilumya (tildrakizumab), Simulect (basiliximab), Zenapax (daclizumab), Spevigo (spesolimab), Cinqair (reslizumab), Kevzara (sarilumab), Sylvant (siltuximab), Adbry (tralokinumab), Ebglyss (lebrikizumab), comprises a heavy chain (HC) comprising mutations, such as those provided for herein, to the Fc domain of

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[0663] IPTS / 200163498.2 DOCKET NO. SES-020WO PATENT the heavy chain that render the Fc domain resistant to proteolytic cleavage, and a light chain (LC) known in the art to be the light chain (LC) of said therapeutic antibody.

[0664] In some embodiments, the therapeutic antibody of Table 10 comprises a heavy chain (HC) amino acid sequence as shown in Table 10, column 4, or a variant thereof. In some embodiments, the therapeutic antibody of Table 10 comprises a heavy chain (HC) amino acid sequence In some embodiments, the heavy chain (HC) amino acid sequence comprises an Fc domain as shown in Table 10, column 4, or a variant thereof. In some embodiments, the therapeutic antibody comprising an Fc domain further comprises a mutation, or at least one mutation, that render the Fc domain resistant to proteolytic cleavage. In some embodiments, the therapeutic antibody comprising an Fc domain further comprises a mutation, or at least one mutation, that render the Fc domain resistant to proteolytic cleavage by a protease, such as those provided herein.

[0665] In some embodiments, a therapeutic antibody comprising an Fc domain comprises a mutation corresponding to Y296Q mutation, according to EU numbering. In some embodiments, a therapeutic antibody comprising an Fc domain comprises a mutation corresponding to F296Q mutation, according to EU numbering. In some embodiments, a therapeutic antibody comprising an Fc domain comprises a mutation corresponding to P329K mutation, according to EU numbering. In some embodiments, a therapeutic antibody comprising an Fc domain comprises a set of mutations corresponding to Y296Q and P329K mutations, according to EU numbering. In some embodiments, a therapeutic antibody comprising an Fc domain comprises a set of mutations corresponding to F296Q and P329K mutations, according to EU numbering.

[0666] In some embodiments, positions 296 and / or 329, according to EU numbering, may be represented as linear positions in the Fc domain amino acid sequence of the therapeutic antibody heavy chain sequence. For example, and without limitation, Rituxan (rituximab) comprises the Fc domain amino acid sequence shown in Table 10, and accordingly, position 296, according to EU numbering, corresponds to position 179 in the linear amino acid sequence of the Fc domain, and position 329, according to EU numbering, corresponds to position 212 in the linear amino acid sequence of the Fc domain, as shown in Table 10. Similar linear representation of each position 296 and / or 329, according to EU numbering, may be conducted for any Fc domain amino acid sequence as shown in Table 10, regardless of immunoglobulin isotype, by assigning position 1 to the first amino acid in the Fc domain sequence as shown in Table 10.

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[0668] IPTS / 200163498.2 DOCKET NO. SES-020WO PATENT

[0669] In some embodiments, the therapeutic antibody selected from Rituxan (rituximab), Ocrevus (ocrelizumab), Orencia (abatacept), Stelara (ustekinumab), Cosentyx (secukinumab), Tremfya (guselkumab), Skyrizi (risankizumab), Actemra (tocilizumab), Enbrel (etanercept), Humira (adalimumab), Benlysta (belimumab), Saphnelo (anifrolumab), Xolair (omalizumab), Bimzelx (bimekizumab), Nucala (mepolizumab), Fasenra (benralizumab), Remicade (infliximab), Simponi (golimumab), Entyvio (vedolizumab), Briumvi (ublituximab), Orthoclone Okt3 (Muromonab-CD3), MabCampath / Lemtrada) (lemtuzumab), Alzumab (itolizumab), Haris (canakinumab), Arcalyst (rilonacept), Ilumya (tildrakizumab), Simulect (basiliximab), Zenapax (daclizumab), Spevigo (spesolimab), Kevzara (sarilumab), Sylvant (siltuximab), any combination thereof, and any variant thereof, comprises a mutation corresponding to Y296Q mutation in the Fc domain, according to EU numbering.

[0670] In some embodiments, the therapeutic antibody selected from Dupixent (dupilumab), Tysabri (natalizumab), Taltz (ixekizumab), Prolia (denosumab), Veopoz (pozelimab), Soliris (eculizumab), Siliq (brodalumab), Omvoh (mirikizumab), Cinqair (reslizumab), Adbry (tralokinumab), Ebglyss (lebrikizumab), and any variant thereof, comprises a mutation corresponding to F296Q mutation in the Fc domain, according to EU numbering.

[0671] In some embodiments, the therapeutic antibody selected from Rituxan (rituximab), Ocrevus (ocrelizumab), Orencia (abatacept), Stelara (ustekinumab), Cosentyx (secukinumab), Tremfya (guselkumab), Skyrizi (risankizumab), Dupixent (dupilumab), Actemra (tocilizumab), Enbrel (etanercept), Humira (adalimumab), Tysabri (natalizumab), Benlysta (belimumab), Saphnelo (anifrolumab), Xolair (omalizumab), Taltz (ixekizumab), Bimzelx (bimekizumab), Nucala (mepolizumab), Fasenra (benralizumab), Prolia (denosumab), Remicade (infliximab), Simponi (golimumab), Entyvio (vedolizumab), Briumvi (ublituximab), Orthoclone Okt3 (Muromonab-CD3), MabCampath / Lemtrada) (lemtuzumab), Alzumab (itolizumab), Veopoz (pozelimab), Soliris (eculizumab), Siliq (brodalumab), Haris (canakinumab), Arcalyst (rilonacept), Omvoh (mirikizumab), Ilumya (tildrakizumab), Simulect (basiliximab), Zenapax (daclizumab), Spevigo (spesolimab), Cinqair (reslizumab), Kevzara (sarilumab), Sylvant (siltuximab), Adbry (tralokinumab), Ebglyss (lebrikizumab), and any variant thereof, comprises a mutation corresponding to P329K mutation in the Fc domain, according to EU numbering.

[0672] In some embodiments, the therapeutic antibody selected from Rituxan (rituximab), Ocrevus (ocrelizumab), Orencia (abatacept), Stelara (ustekinumab), Cosentyx (secukinumab),

[0673] -137-

[0674] IPTS / 200163498.2 DOCKET NO. SES-020WO PATENT

[0675] Tremfya (guselkumab), Skyrizi (risankizumab), Actemra (tocilizumab), Enbrel (etanercept), Humira (adalimumab), Benlysta (belimumab), Saphnelo (anifrolumab), Xolair (omalizumab), Bimzelx (bimekizumab), Nucala (mepolizumab), Fasenra (benralizumab), Remicade (infliximab), Simponi (golimumab), Entyvio (vedolizumab), Briumvi (ublituximab), Orthoclone Okt3 (Muromonab-CD3), MabCampath / Lemtrada) (lemtuzumab), Alzumab (itolizumab), Haris (canakinumab), Arcalyst (rilonacept), Ilumya (tildrakizumab), Simulect (basiliximab), Zenapax (daclizumab), Spevigo (spesolimab), Kevzara (sarilumab), Sylvant (siltuximab), and any variant thereof, comprises a set of mutations corresponding to Y296Q and P329K mutations in the Fc domain, according to EU numbering.

[0676] In some embodiments, the therapeutic antibody selected from Dupixent (dupilumab), Tysabri (natalizumab), Taltz (ixekizumab), Prolia (denosumab), Veopoz (pozelimab), Soliris (eculizumab), Siliq (brodalumab), Omvoh (mirikizumab), Cinqair (reslizumab), Adbry (tralokinumab), and Ebglyss (lebrikizumab), and any variant thereof, comprises a set of mutations corresponding to F296Q and P329K mutations in the Fc domain, according to EU numbering.

[0677] In some embodiments, the therapeutic antibody selected from Rituxan (rituximab), Ocrevus (ocrelizumab), Orencia (abatacept), Stelara (ustekinumab), Cosentyx (secukinumab), Tremfya (guselkumab), Skyrizi (risankizumab), Actemra (tocilizumab), Enbrel (etanercept), Humira (adalimumab), Benlysta (belimumab), Saphnelo (anifrolumab), Xolair (omalizumab), Bimzelx (bimekizumab), Nucala (mepolizumab), Fasenra (benralizumab), Remicade (infliximab), Simponi (golimumab), Entyvio (vedolizumab), Briumvi (ublituximab), Orthoclone Okt3 (Muromonab-CD3), MabCampath / Lemtrada) (lemtuzumab), Alzumab (itolizumab), Haris (canakinumab), Arcalyst (rilonacept), Ilumya (tildrakizumab), Simulect (basiliximab), Zenapax (daclizumab), Spevigo (spesolimab), Kevzara (sarilumab), Sylvant (siltuximab), and any variant thereof, comprises a heavy chain comprising a mutation corresponding to Y296Q mutation in the Fc domain of the heavy chain, according to EU numbering.

[0678] In some embodiments, the therapeutic antibody Rituxan (rituximab) comprises a heavy chain comprising a mutation corresponding to Y296Q mutation in the Fc domain of the heavy chain, according to EU numbering. In some embodiments, the therapeutic antibody Ocrevus (ocrelizumab) comprises a heavy chain comprising a mutation corresponding to Y296Q mutation in the Fc domain of the heavy chain, according to EU numbering. In some embodiments, the

[0679] -138-

[0680] IPTS / 200163498.2 DOCKET NO. SES-020WO PATENT therapeutic antibody Orencia (abatacept) comprises a heavy chain comprising a mutation corresponding to Y296Q mutation in the Fc domain of the heavy chain, according to EU numbering. In some embodiments, the therapeutic antibody Stelara (ustekinumab) comprises a heavy chain comprising a mutation corresponding to Y296Q mutation in the Fc domain of the heavy chain, according to EU numbering. In some embodiments, the therapeutic antibody Cosentyx (secukinumab) comprises a heavy chain comprising a mutation corresponding to Y296Q mutation in the Fc domain of the heavy chain, according to EU numbering. In some embodiments, the therapeutic antibody Tremfya (guselkumab) comprises a heavy chain comprising a mutation corresponding to Y296Q mutation in the Fc domain of the heavy chain, according to EU numbering. In some embodiments, the therapeutic antibody Skyrizi (risankizumab) comprises a heavy chain comprising a mutation corresponding to Y296Q mutation in the Fc domain of the heavy chain, according to EU numbering. In some embodiments, the therapeutic antibody Actemra (tocilizumab) comprises a heavy chain comprising a mutation corresponding to Y296Q mutation in the Fc domain of the heavy chain, according to EU numbering. In some embodiments, the therapeutic antibody Enbrel (etanercept) comprises a heavy chain comprising a mutation corresponding to Y296Q mutation in the Fc domain of the heavy chain, according to EU numbering. In some embodiments, the therapeutic antibody Humira (adalimumab) comprises a heavy chain comprising a mutation corresponding to Y296Q mutation in the Fc domain of the heavy chain, according to EU numbering. In some embodiments, the therapeutic antibody Benlysta (belimumab) comprises a heavy chain comprising a mutation corresponding to Y296Q mutation in the Fc domain of the heavy chain, according to EU numbering. In some embodiments, the therapeutic antibody Saphnelo (anifrolumab) comprises a heavy chain comprising a mutation corresponding to Y296Q mutation in the Fc domain of the heavy chain, according to EU numbering. In some embodiments, the therapeutic antibody Xolair (omalizumab) comprises a heavy chain comprising a mutation corresponding to Y296Q mutation in the Fc domain of the heavy chain, according to EU numbering. In some embodiments, the therapeutic antibody Bimzelx (bimekizumab) comprises a heavy chain comprising a mutation corresponding to Y296Q mutation in the Fc domain of the heavy chain, according to EU numbering. In some embodiments, the therapeutic antibody Nucala (mepolizumab) comprises a heavy chain comprising a mutation corresponding to Y296Q mutation in the Fc domain of the heavy chain, according to EU numbering. In some

[0681] -139-

[0682] ...

Claims

DOCKET NO. SES-020WO PATENTWhat is claimed:

1. An immunoglobulin protease comprising an amino acid sequence having at least 90% identity to SEQ ID NO: 35, and having a set of mutations selected from: an aspartic acid (D) at position 163; a threonine (T) at position 60, an aspartic acid (D) at position 61, an isoleucine (I) at position 63, a tyrosine (Y) at position 64, a leucine (L) at position 69, a lysine (K) at position 74, a glycine (G) at position 76, a leucine (L) at position 85, a glutamic acid (E) at position 112, an asparagine (N) at position 116, a lysine (K) at position 119, a lysine (K) at position 120, a tyrosine (Y) at position 121, a glycine (G) at position 130, a leucine (L) at position 134, an aspartic acid (D) at position 145, a serine (S) at position 146, a threonine (T) at position 148, an aspartic acid (D) at position 154, an aspartic acid (D) at position 190, an arginine (R) at position 198, an asparagine (N) at position 223, a tyrosine (Y) at position 225, a leucine (L) at position 227, a threonine (T) at position 231, a lysine (K) at position 241, an isoleucine (I) at position 251, a glycine (G) at position 261, an alanine (A) at position 273, a glutamic acid (E) at position 274, a glutamic acid (E) at position 278, a proline (P) at position 289, a leucine (L) at position 303, a glutamic acid (E) at position 314, and an isoleucine (I) at position 334, wherein the positions correspond to the numbering as set forth in SEQ ID NO: 35; a threonine (T) at position 60, an aspartic acid (D) at position 61, an isoleucine (I) at position 63, a tyrosine (Y) at position 64, a leucine (L) at position 69, a lysine (K) at position 74, a glycine (G) at position 76, a leucine (L) at position 85, a glutamic acid (E) at position 112, an asparagine (N) at position 116, a lysine (K) at position 119, a lysine (K) at position 120, a tyrosine (Y) at position 121, a glycine (G) at position 130, a leucine (L) at position 134, an aspartic acid (D) at position 145, a serine (S) at position 146, a threonine (T) at position 148, an aspartic acid (D) at position 154, an aspartic acid (D) at position 163, an aspartic acid (D) at position 190, an arginine (R) at position 198, an asparagine (N) at position 223, a tyrosine (Y) at position 225, a leucine (L) at position 227, a threonine (T) at position 231, a lysine (K) at position 241, an isoleucine (I) at position 251, a glycine (G) at position 261, an alanine (A) at position 273, a glutamic acid (E) at position 274, a glutamic acid (E) at position 278, a proline (P) at position 289, a leucine (L) at position 303, a glutamic acid (E) at position 314, and an-240-IPTS / 200163498.2DOCKET NO. SES-020WO PATENT isoleucine (I) at position 334, wherein the positions correspond to the numbering as set forth in SEQ ID NO: 35; a threonine (T) at position 60, an aspartic acid (D) at position 61, an isoleucine (I) at position 63, a tyrosine (Y) at position 64, a leucine (L) at position 69, a lysine (K) at position 74, a glycine (G) at position 76, a leucine (L) at position 85, a glutamic acid (E) at position 112, an asparagine (N) at position 116, a lysine (K) at position 119, a lysine (K) at position 120, a tyrosine (Y) at position 121, a glycine (G) at position 130, a leucine (L) at position 134, an aspartic acid (D) at position 145, a serine (S) at position 146, a threonine (T) at position 148, an aspartic acid (D) at position 154, an asparagine (N) at position 163, an alanine (A) at position 166, an arginine (R) at position 167, a glutamine (Q) at position 168, an aspartic acid (D) at position 190, an arginine (R) at position 198, an asparagine (N) at position 223, a tyrosine (Y) at position 225, a leucine (L) at position 227, a threonine (T) at position 231, a lysine (K) at position 241, an isoleucine (I) at position 251, a glycine (G) at position 261, an alanine (A) at position 273, a glutamic acid (E) at position 274, a glutamic acid (E) at position 278, a proline (P) at position 289, a leucine (L) at position 303, a glutamic acid (E) at position 314, and an isoleucine (I) at position 334, wherein the positions correspond to the numbering as set forth in SEQ ID NO: 35; or a threonine (T) at position 60, an aspartic acid (D) at position 61, an isoleucine (I) at position 63, a tyrosine (Y) at position 64, a leucine (L) at position 69, a lysine (K) at position 74, a glycine (G) at position 76, a leucine (L) at position 85, a glutamic acid (E) at position 112, an asparagine (N) at position 116, a lysine (K) at position 119, a lysine (K) at position 120, a tyrosine (Y) at position 121, a glycine (G) at position 130, a leucine (L) at position 134, an aspartic acid (D) at position 145, a serine (S) at position 146, a threonine (T) at position 148, an aspartic acid (D) at position 154, an asparagine (N) at position 163, an alanine (A) at position 166, an arginine (R) at position 167, a glutamine (Q) at position 168, an isoleucine (I) at position 169, an aspartic acid (D) at position 190, an arginine (R) at position 198, an asparagine (N) at position 223, a tyrosine (Y) at position 225, a leucine (L) at position 227, a threonine (T) at position 231, a lysine (K) at position 241, an isoleucine (I) at position 251, a glycine (G) at position 261, an alanine (A) at position 273, a glutamic acid (E) at position 274, a glutamic acid (E) at position 278, a proline (P) at position 289, a leucine (L) at position 303, a glutamic acid-241-IPTS / 200163498.2DOCKET NO. SES-020WO PATENT(E) at position 314, and an isoleucine (I) at position 334, wherein the positions correspond to the numbering as set forth in SEQ ID NO: 35.

2. The immunoglobulin protease of claim 1, wherein the immunoglobulin protease has protease activity that is specific for immunoglobulins.

3. The immunoglobulin protease of any one of claims 1 or 2, wherein the protease activity is IgG cleaving protease activity.

4. The immunoglobulin protease of any one of claims 1-3, wherein the immunoglobulin protease has the same or improved protease activity as compared to a wild-type IdeS protease.

5. The immunoglobulin protease of any one of claims 1-4, wherein the immunoglobulin protease comprises an amino acid sequence having at least 90% to any one of SEQ ID NOs: 57, 58, 59, or 60.

6. The immunoglobulin protease of claim 5, wherein the immunoglobulin protease comprises the amino acid sequence of any one of SEQ ID NOs: 57, 58, 59, or 60.

7. The immunoglobulin protease of any one of claims 1-6, wherein the immunoglobulin protease is covalently or non-covalently conjugated to an Fc polypeptide.

8. The immunoglobulin protease of claim 7, wherein the immunoglobulin protease is conjugated to the Fc polypeptide via a peptide linker.

9. The immunoglobulin protease of claim 8, wherein the peptide linker is a glycine / serine linker.

10. The immunoglobulin protease of claim 7, wherein the Fc polypeptide comprises an amino acid sequence of any one of SEQ ID NO: 11, 12, 13, 19, 20, 21, 22, 23, 24, 25, 42, 43, 44, 45, 46, 47, 48, 49, 200, 201, or 202.-242-IPTS / 200163498.2DOCKET NO. SES-020WO PATENT11. The immunoglobulin protease of any of claim 1-10, wherein the immunoglobulin protease does not comprise an amino acid sequence of any one of SEQ ID NO: 15, 16, 17, 18, 27, 30, 32, 33, 34, 53, 54, 56, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, or 106.

12. A molecule comprising: i) an immunoglobulin protease comprising an amino acid sequence having at least 90% identity to:SEQ ID NO: 57, provided that the immunoglobulin protease comprises a threonine (T) at position 60, an aspartic acid (D) at position 61, an isoleucine (I) at position 63, a tyrosine (Y) at position 64, a leucine (L) at position 69, a lysine (K) at position 74, a glycine (G) at position 76, a leucine (L) at position 85, a glutamic acid (E) at position 112, an asparagine (N) at position 116, a lysine (K) at position 119, a lysine (K) at position 120, a tyrosine (Y) at position 121, a glycine (G) at position 130, a leucine (L) at position 134, an aspartic acid (D) at position 145, a serine (S) at position 146, a threonine (T) at position 148, an aspartic acid (D) at position 154, an aspartic acid (D) at position 190, an arginine (R) at position 198, an asparagine (N) at position 223, a tyrosine (Y) at position 225, a leucine (L) at position 227, a threonine (T) at position 231, a lysine (K) at position 241, an isoleucine (I) at position 251, a glycine (G) at position 261, an alanine (A) at position 273, a glutamic acid (E) at position 274, a glutamic acid (E) at position 278, a proline (P) at position 289, a leucine (L) at position 303, a glutamic acid (E) at position 314, and an isoleucine (I) at position 334, wherein the positions correspond to the numbering as set forth in SEQ ID NO: 35;SEQ ID NO: 58, provided that the immunoglobulin protease comprises a threonine (T) at position 60, an aspartic acid (D) at position 61, an isoleucine (I) at position 63, a tyrosine (Y) at position 64, a leucine (L) at position 69, a lysine (K) at position 74, a glycine (G) at position 76, a leucine (L) at position 85, a glutamic acid (E) at position 112, an asparagine (N) at position 116, a lysine (K)-243-IPTS / 200163498.2DOCKET NO. SES-020WO PATENT at position 119, a lysine (K) at position 120, a tyrosine (Y) at position 121, a glycine (G) at position 130, a leucine (L) at position 134, an aspartic acid (D) at position 145, a serine (S) at position 146, a threonine (T) at position 148, an aspartic acid (D) at position 154, an aspartic acid (D) at position 163, an aspartic acid (D) at position 190, an arginine (R) at position 198, an asparagine (N) at position 223, a tyrosine (Y) at position 225, a leucine (L) at position 227, a threonine (T) at position 231, a lysine (K) at position 241, an isoleucine (I) at position 251, a glycine (G) at position 261, an alanine (A) at position 273, a glutamic acid (E) at position 274, a glutamic acid (E) at position 278, a proline (P) at position 289, a leucine (L) at position 303, a glutamic acid (E) at position 314, and an isoleucine (I) at position 334, wherein the positions correspond to the numbering as set forth in SEQ ID NO: 35;SEQ ID NO: 59, provided that the immunoglobulin protease comprises a threonine (T) at position 60, an aspartic acid (D) at position 61, an isoleucine (I) at position 63, a tyrosine (Y) at position 64, a leucine (L) at position 69, a lysine (K) at position 74, a glycine (G) at position 76, a leucine (L) at position 85, a glutamic acid (E) at position 112, an asparagine (N) at position 116, a lysine (K) at position 119, a lysine (K) at position 120, a tyrosine (Y) at position 121, a glycine (G) at position 130, a leucine (L) at position 134, an aspartic acid (D) at position 145, a serine (S) at position 146, a threonine (T) at position 148, an aspartic acid (D) at position 154, an asparagine (N) at position 163, an alanine (A) at position 166, an arginine (R) at position 167, a glutamine (Q) at position 168, an aspartic acid (D) at position 190, an arginine (R) at position 198, an asparagine (N) at position 223, a tyrosine (Y) at position 225, a leucine (L) at position 227, a threonine (T) at position 231, a lysine (K) at position 241, an isoleucine (I) at position 251, a glycine (G) at position 261, an alanine (A) at position 273, a glutamic acid (E) at position 274, a glutamic acid (E) at position 278, a proline (P) at position 289, a leucine (L) at position 303, a glutamic acid (E) at position 314, and an isoleucine (I) at position 334, wherein the positions correspond to the numbering as set forth in SEQ ID NO: 35; or-244-IPTS / 200163498.2DOCKET NO. SES-020WO PATENTSEQ ID NO: 60, provided that the immunoglobulin protease comprises a threonine (T) at position 60, an aspartic acid (D) at position 61, an isoleucine (I) at position 63, a tyrosine (Y) at position 64, a leucine (L) at position 69, a lysine (K) at position 74, a glycine (G) at position 76, a leucine (L) at position 85, a glutamic acid (E) at position 112, an asparagine (N) at position 116, a lysine (K) at position 119, a lysine (K) at position 120, a tyrosine (Y) at position 121, a glycine (G) at position 130, a leucine (L) at position 134, an aspartic acid (D) at position 145, a serine (S) at position 146, a threonine (T) at position 148, an aspartic acid (D) at position 154, an asparagine (N) at position 163, an alanine (A) at position 166, an arginine (R) at position 167, a glutamine (Q) at position 168, an isoleucine (I) at position 169, an aspartic acid (D) at position 190, an arginine (R) at position 198, an asparagine (N) at position 223, a tyrosine (Y) at position 225, a leucine (L) at position 227, a threonine (T) at position 231, a lysine (K) at position 241, an isoleucine (I) at position 251, a glycine (G) at position 261, an alanine (A) at position 273, a glutamic acid (E) at position 274, a glutamic acid (E) at position 278, a proline (P) at position 289, a leucine (L) at position 303, a glutamic acid (E) at position 314, and an isoleucine (I) at position 334, wherein the positions correspond to the numbering as set forth in SEQ ID NO: 35; and ii) an Fc polypeptide, wherein the immunoglobulin protease is conjugated to the Fc polypeptide via a linker.

13. The molecule of claim 12, wherein the immunoglobulin protease further comprises a mutation of tryptophan (W) at position 155, arginine (R) at position 155, leucine (L) at position 157, lysine (K) at position 158, leucine (L) at position 240, arginine (R) at position 238, glutamine (Q) at position 241, glycine (G) at position 258, lysine (K) at position 258, leucine (L) at position 260, or any combination thereof, as compared to SEQ ID NO: 35.

14. The molecule of any one of claims 12 or 13, wherein the immunoglobulin protease has protease activity.-245-IPTS / 200163498.2DOCKET NO. SES-020WO PATENT15. The molecule of any one of claims 12-14, wherein the immunoglobulin protease having protease activity is an immunoglobulin protease having IgG cleaving protease activity.

16. The molecule of any one of claims 12-15, wherein the immunoglobulin protease has the same or improved protease activity as compared to a wild-type IdeS protease.

17. The molecule of claim 12, wherein the immunoglobulin protease comprises the amino acid sequence of any one of SEQ ID NOs: 57, 58, 59, or 60.

18. The molecule of claim 12, wherein the linker is a peptide linker.

19. The molecule of claim 18, wherein the peptide linker is a glycine / serine linker.

20. The molecule of claim 12, wherein the Fc polypeptide comprises any one of SEQ ID NO:11, 12, 13, 19, 20, 21, 22, 23, 24, 25, 42, 43, 44, 45, 46, 47, 48, 49, 200, 201, or 202.

21. The molecule of any one of claims 12-20, wherein the molecule comprises: i) the immunoglobulin protease comprising the amino acid sequence having at least 90% identity to SEQ ID NO: 57, provided that the immunoglobulin protease comprises a threonine (T) at position 60, an aspartic acid (D) at position 61, an isoleucine (I) at position 63, a tyrosine (Y) at position 64, a leucine (L) at position 69, a lysine (K) at position 74, a glycine (G) at position 76, a leucine (L) at position 85, a glutamic acid (E) at position 112, an asparagine (N) at position 116, a lysine (K) at position 119, a lysine(K) at position 120, a tyrosine (Y) at position 121, a glycine (G) at position 130, a leucine(L) at position 134, an aspartic acid (D) at position 145, a serine (S) at position 146, a threonine (T) at position 148, an aspartic acid (D) at position 154, an aspartic acid (D) at position 190, an arginine (R) at position 198, an asparagine (N) at position 223, a tyrosine (Y) at position 225, a leucine (L) at position 227, a threonine (T) at position 231, a lysine (K) at position 241, an isoleucine (I) at position 251, a glycine (G) at position 261, an alanine (A) at position 273, a glutamic acid (E) at position 274, a glutamic acid (E) at position 278, a proline (P) at position 289, a leucine (L) at position 303, a glutamic acid-246-IPTS / 200163498.2DOCKET NO. SES-020WO PATENT(E) at position 314, and an isoleucine (I) at position 334, wherein the positions correspond to the numbering as set forth in SEQ ID NO: 35; and the Fc polypeptide having the amino acid sequence of SEQ ID NO: 23; ii) the immunoglobulin protease comprising the amino acid sequence having at least 90 identity to SEQ ID NO: 58, provided that the immunoglobulin protease comprises a threonine (T) at position 60, an aspartic acid (D) at position 61, an isoleucine (I) at position 63, a tyrosine (Y) at position 64, a leucine (L) at position 69, a lysine (K) at position 74, a glycine (G) at position 76, a leucine (L) at position 85, a glutamic acid (E) at position 112, an asparagine (N) at position 116, a lysine (K) at position 119, a lysine(K) at position 120, a tyrosine (Y) at position 121, a glycine (G) at position 130, a leucine(L) at position 134, an aspartic acid (D) at position 145, a serine (S) at position 146, a threonine (T) at position 148, an aspartic acid (D) at position 154, an aspartic acid (D) at position 163, an aspartic acid (D) at position 190, an arginine (R) at position 198, an asparagine (N) at position 223, a tyrosine (Y) at position 225, a leucine (L) at position 227, a threonine (T) at position 231, a lysine (K) at position 241, an isoleucine (I) at position 251, a glycine (G) at position 261, an alanine (A) at position 273, a glutamic acid (E) at position 274, a glutamic acid (E) at position 278, a proline (P) at position 289, a leucine (L) at position 303, a glutamic acid (E) at position 314, and an isoleucine (I) at position 334, wherein the positions correspond to the numbering as set forth in SEQ ID NO: 35; and the Fc polypeptide having the amino acid sequence of SEQ ID NO: 23; iii) the immunoglobulin protease comprising the amino acid sequence having at least 90% identity to SEQ ID NO: 59, provided that the immunoglobulin protease comprises a threonine (T) at position 60, an aspartic acid (D) at position 61, an isoleucine (I) at position 63, a tyrosine (Y) at position 64, a leucine (L) at position 69, a lysine (K) at position 74, a glycine (G) at position 76, a leucine (L) at position 85, a glutamic acid (E) at position 112, an asparagine (N) at position 116, a lysine (K) at position 119, a lysine(K) at position 120, a tyrosine (Y) at position 121, a glycine (G) at position 130, a leucine(L) at position 134, an aspartic acid (D) at position 145, a serine (S) at position 146, a threonine (T) at position 148, an aspartic acid (D) at position 154, an asparagine (N) at position 163, an alanine (A) at position 166, an arginine (R) at position 167, a glutamine-247-IPTS / 200163498.2DOCKET NO. SES-020WO PATENT(Q) at position 168, an aspartic acid (D) at position 190, an arginine (R) at position 198, an asparagine (N) at position 223, a tyrosine (Y) at position 225, a leucine (L) at position 227, a threonine (T) at position 231, a lysine (K) at position 241, an isoleucine (I) at position 251, a glycine (G) at position 261, an alanine (A) at position 273, a glutamic acid (E) at position 274, a glutamic acid (E) at position 278, a proline (P) at position 289, a leucine (L) at position 303, a glutamic acid (E) at position 314, and an isoleucine (I) at position 334, wherein the positions correspond to the numbering as set forth in SEQ ID NO: 35; and the Fc polypeptide having the amino acid sequence of SEQ ID NO: 23; iv) the immunoglobulin protease comprising the amino acid sequence having at least 90% identity to SEQ ID NO: 60, provided that the immunoglobulin protease comprises a threonine (T) at position 60, an aspartic acid (D) at position 61, an isoleucine (I) at position 63, a tyrosine (Y) at position 64, a leucine (L) at position 69, a lysine (K) at position 74, a glycine (G) at position 76, a leucine (L) at position 85, a glutamic acid (E) at position 112, an asparagine (N) at position 116, a lysine (K) at position 119, a lysine(K) at position 120, a tyrosine (Y) at position 121, a glycine (G) at position 130, a leucine(L) at position 134, an aspartic acid (D) at position 145, a serine (S) at position 146, a threonine (T) at position 148, an aspartic acid (D) at position 154, an asparagine (N) at position 163, an alanine (A) at position 166, an arginine (R) at position 167, a glutamine (Q) at position 168, an isoleucine (I) at position 169, an aspartic acid (D) at position 190, an arginine (R) at position 198, an asparagine (N) at position 223, a tyrosine (Y) at position 225, a leucine (L) at position 227, a threonine (T) at position 231, a lysine (K) at position 241, an isoleucine (I) at position 251, a glycine (G) at position 261, an alanine (A) at position 273, a glutamic acid (E) at position 274, a glutamic acid (E) at position 278, a proline (P) at position 289, a leucine (L) at position 303, a glutamic acid (E) at position 314, and an isoleucine (I) at position 334, wherein the positions correspond to the numbering as set forth in SEQ ID NO: 35; and the Fc polypeptide having the amino acid sequence of SEQ ID NO: 23; or v) the immunoglobulin protease comprising the amino acid sequence having at least 90% identity to SEQ ID NO: 60, provided that the immunoglobulin protease comprises a threonine (T) at position 60, an aspartic acid (D) at position 61, an isoleucine-248-IPTS / 200163498.2DOCKET NO. SES-020WO PATENT(T) at position 63, a tyrosine (Y) at position 64, a leucine (L) at position 69, a lysine (K) at position 74, a glycine (G) at position 76, a leucine (L) at position 85, a glutamic acid (E) at position 112, an asparagine (N) at position 116, a lysine (K) at position 119, a lysine(K) at position 120, a tyrosine (Y) at position 121, a glycine (G) at position 130, a leucine(L) at position 134, an aspartic acid (D) at position 145, a serine (S) at position 146, a threonine (T) at position 148, an aspartic acid (D) at position 154, an asparagine (N) at position 163, an alanine (A) at position 166, an arginine (R) at position 167, a glutamine (Q) at position 168, an isoleucine (I) at position 169, an aspartic acid (D) at position 190, an arginine (R) at position 198, an asparagine (N) at position 223, a tyrosine (Y) at position 225, a leucine (L) at position 227, a threonine (T) at position 231, a lysine (K) at position 241, an isoleucine (I) at position 251, a glycine (G) at position 261, an alanine (A) at position 273, a glutamic acid (E) at position 274, a glutamic acid (E) at position 278, a proline (P) at position 289, a leucine (L) at position 303, a glutamic acid (E) at position 314, and an isoleucine (I) at position 334, wherein the positions correspond to the numbering as set forth in SEQ ID NO: 35; and the Fc polypeptide having the amino acid sequence of SEQ ID NO: 24.

22. The molecule of any one of claims 12-21, wherein the molecule comprises an amino acid sequence of any one of SEQ ID NO: 114, 115, 116, 117, or 118.

23. The molecule of any one of claims 12-22, wherein the Fc polypeptide self-associates, or associates with another polypeptide, to form a dimer.

24. The molecule of claim 23, wherein the dimer is a homodimer or a heterodimer.

25. The molecule of any one of claims 12-24, wherein the molecule is a monovalent molecule or a bivalent molecule.

26. The molecule of any of claim 12-25, wherein the immunoglobulin protease does not comprise an amino acid sequence of any one of SEQ ID NO: 15, 16, 17, 18, 27, 30, 32, 33, 34,-249-IPTS / 200163498.2DOCKET NO. SES-020WO PATENT53, 54, 56, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71 , 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, or 106.

27. A molecule comprising: an immunoglobulin protease comprising an amino acid sequence of any one of SEQ ID NO: 57, 58, 59, or 60; and an Fc polypeptide, wherein the immunoglobulin protease is conjugated to the Fc polypeptide via a linker, such as a peptide linker.

28. The molecule of claim 27, wherein the immunoglobulin protease has protease activity.

29. The molecule of any one of claims 27 or 28, wherein the immunoglobulin protease having protease activity is an immunoglobulin protease having IgG cleaving protease activity.

30. The molecule of any one of claims 27-29, wherein the immunoglobulin protease has the same or improved protease activity as compared to a wild-type IdeS protease.

31. The molecule of claim 30, wherein the peptide linker is a glycine / serine linker.

32. The molecule of claim 31, wherein the glycine / serine linker has the amino acid sequence of SEQ ID NO: 121.

33. The molecule of any one of claims 27-32, wherein the Fc polypeptide comprises any one of SEQ ID NO: 11, 12, 13, 19, 20, 21, 22, 23, 24, 25, 42, 43, 44, 45, 46, 47, 48, 49, 200, 201, or 202.

34. The molecule of any one of claims 27-33, wherein the molecule comprises: i) the immunoglobulin protease having the amino acid sequence of amino acid sequence SEQ ID NO: 57; and the Fc polypeptide having the amino acid sequence of SEQ ID NO: 23;-250-IPTS / 200163498.2DOCKET NO. SES-020WO PATENT ii) the immunoglobulin protease having the amino acid sequence of amino acid sequence SEQ ID NO: 58; and the Fc polypeptide having the amino acid sequence of SEQ ID NO: 23; iii) the immunoglobulin protease having the amino acid sequence of amino acid sequence SEQ ID NO: 59; and the Fc polypeptide having the amino acid sequence of SEQ ID NO: 23; iv) the immunoglobulin protease having the amino acid sequence of amino acid sequence SEQ ID NO: 60; and the Fc polypeptide having the amino acid sequence of SEQ ID NO: 23; or v) the immunoglobulin protease having the amino acid sequence of amino acid sequence SEQ ID NO: 60; and the Fc polypeptide having the amino acid sequence of SEQ ID NO: 24.

35. The molecule of any one of claims 27-34, wherein the molecule comprises the amino acid sequence of any one of SEQ ID NO: 114, 115, 116, 117, or 118.

36. The molecule of any one of claims 27-35, wherein the Fc polypeptide self-associates, or associates with another polypeptide, to form a dimer.

37. The molecule of claim 36, wherein the dimer is a homodimer or a heterodimer.

38. The molecule of any one of claims 27-37, wherein the molecule is a monovalent molecule or a bivalent molecule.

39. The molecule of any of claim 27-38, wherein the immunoglobulin protease does not comprise an amino acid sequence of any one of SEQ ID NO: 15, 16, 17, 18, 27, 30, 32, 33, 34, 53, 54, 56, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, or 106.

40. A pharmaceutical composition comprising the immunoglobulin protease of any one of claims 1-11, or the molecule of any one of claims 12-39.-251-IPTS / 200163498.2DOCKET NO. SES-020WO PATENT41. A pharmaceutical composition comprising an IgG protease variant comprising a Y136D substitution as compared, wherein the positions correspond to the numbering as set forth in SEQ ID NO: 35, wherein the IgG protease aggregates less as compared to an IgG protease without the Y136D mutation.

42. The pharmaceutical composition of claim 41, wherein the pharmaceutical composition comprises less than 5% high molecular weight complexes of the IgG protease.

43. The pharmaceutical composition of claims 41 or 42, wherein the protease that comprises the Y136D mutation comprises an amino acid sequence that is at least 70% identical to the amino acid sequence of SEQ ID NO: 35 or any other IgG protease.

44. A method of treating a disease or disorder in a subject, the method comprising administering the immunoglobulin protease of any one of claims 1-11, the molecule of any one of claims 12-39, or the pharmaceutical compositions of any one of claims 40-43, to the subject to treat the disease or disorder.

45. The method of claim 44, wherein the disease or disorder is selected from Addison's disease, Anti-GBM glomerulonephritis, anti -neutrophil cytoplasmic antibody-associated vasculitis, ANCA associated vasculitis, granulomatous polyangiitis (Wegener granulomatosis), eosinophilic granulomatosis with polyangiitis (Churg-Strauss syndrome), microscopic polyangiitis, anti-NMDAR encephalitis, anti-phospholipid antibody syndrome (APS) and catastrophic APS, autoimmune bullous skin diseases, pemphigus, pemphigus foliaceus (PF), fogo selvage (FS), pemphigus vulgaris (PV), autoimmune hemolytic anemia (AIHA), autoimmune hepatitis (AIH), autoimmune neutropenia (AIN), bullous pemphigoid (BP), Celiac’s disease, chronic urticaria, complete congenital heart block (CCHB), diabetes type 1A (T1DM), epidermolysis bullosa acquisita (EBA), essential mixed cryoglobulinemia, Goodpasture’s syndrome, anti-glomerular basement membrane disease, Graves’ disease, Goitre, hyperthyroidism, infiltrative exophthalmos, infiltrative dermopathy, Guillain-Barre syndrome (GBS), acute inflammatory demyelinating polyneuropathy (AIDP), acute motor axonal-252-IPTS / 200163498.2DOCKET NO. SES-020WO PATENT neuropathy (AMAN), hemophilia, acquired FVIT deficiency, idiopathic thrombocytopenic purpura (ITP), Lambert-Eaton myasthenic syndrome (LEMS), mixed connective tissue disease (MCTD), multiple myeloma, myasthenia gravis, myasthenic crisis, myocarditis, dilated cardiomyopathy (DCM), congestive cardiomyopathy, neuromyelitis optica (NMO), primary biliary cirrhosis (PBC), primary progressive multiple sclerosis (PPMS), relapsing-rheumatic multiple sclerosis, acute exacerbation of multiple sclerosis, rheumatic heart disease (RHD), rheumatoid arthritis (RA), serum-sickness, immune complex hypersensitivity (type III), Sjogren’s syndrome (SS), lupus nephritis, systemic lupus erythematosus (SLE), cutaneous lupus (CLE), transplant rejection, thrombotic thrombocytopenic purpura (TTP), idiopathic inflammatory myopathies, polymyositis, dermatomyositis, inclusion body myositis, uveitis, scleritis, ankylosing spondylitis, axial spondyloarthritis, reactive arthritis, psoriatic arthritis, IBD- associated arthritis, antiphospholipid syndrome, systemic sclerosis, giant cell arteritis, polymyalgia rheumatic, Takayasu’s arteritis, anti-neutrophil cytoplasmic antibody associated vasculitis, Henoch-Schonlein purpura, polyarteritis nodosa, Cogan’s syndrome, Buerger’s disease, Susan’s disease, immune complex vasculitis, primary angiitis of the central nervous system (CNS), Behcet’s disease, relapsing polychondritis, juvenile idiopathic arthritis, juvenile dermatomyositis, autoimmune brain disease, sarcoidosis, neurosarcoidosis, IgG4-related diseases, autoimmune complications of immune checkpoint inhibitors (IRAEs), adult onset Still’s disease, warm antibody hemolytic anemia (wAIHA), immune thrombocytopenia, immune thrombotic thrombocytopenia, thrombic thrombocytopenia, pernicious anemia, aplastic anemia, Evan’s syndrome, autoimmune neutropenia, acquired von Willibrand syndrome, recurring fetal loss, Rh mismatch, ulcerative colitis, autoimmune hepatitis, autoimmune pancreatitis, eosinophilic esophagitis / gastritis, primary sclerosing cholangitis, primary biliary sclerosis, glomerulonephritis, glomerular basement membrane disease, scleritis / episcleritis, uveitis, conjunctivitis, keratitis, type I diabetes mellitus, Hashimoto’s thyroiditis, polyglandular autoimmune endocrine syndromes, atopic dermatitis, dermatitis herpetiformis, pemphigus foliaceus, fogo selvagem, cutaneous lupus erythematosus, linear IgA disease, Lichen planus, transplantation, antibody-mediated rejection, alloantibody hypersensitization, xenoantibody mediated rejection, solid organ rejection, graft vs. host disease (GVHD), multiple sclerosis, neuromyelitis optica, amyotrophic lateral sclerosis, Parkinson’s disease, autoimmune encephalitis, CNS vasculitis, chronic idiopathic demyelinating polyneuropathy, transverse-253-IPTS / 200163498.2DOCKET NO. SES-020WO PATENT myelitis, optic neuritis, anti-myelin oligodendrocyte glycoprotein (MOG) disease, chronic meningitis, rheumatic heart disease, infectious disease, vaccination, antibody dependent enhancement, antibody to therapeutic biologic agents (cytokines, monoclonal antibodies, enzymes, coagulation factors), allergic asthma, eosinophilic pneumonia, nonspecific interstitial pneumonia, rheumatoid arthritis-associated interstitial lung disease (RA-ILD), sarcoidosis, hypersensitive pneumonitis, allergic bronchopulmonary mycosis, chronic rhinosinusitis with nasal polyps, COVID-19, long COVID- 19, or any combination thereof.

46. A method of treating a transplant subject comprising administering a therapeutically effective amount of the immunoglobulin protease of any one of claims 1-11, the molecule of any one of claims 12-39, or the pharmaceutical composition of any one of claims 40-43, to the subject, thereby treating the transplant (recipient) subject.

47. A method of improving a gene-therapy treatment in subject, such as reducing antibodies against a gene therapy vector, comprising administering a therapeutically effective amount of the immunoglobulin protease of any one of claims 1-11, the molecule of any one of claims 12-39, or the pharmaceutical composition of any one of claims 40-43, to the subject, prior to, simultaneously with, or after the administration of the gene therapy vector, thereby improving the gene-therapy treatment in the subject.

48. A method of treating a subject having, or at risk, or elevated risk, for having, an IgG mediated disease or disorder, comprising administering a therapeutically effective amount of the immunoglobulin protease of any one of claims 1-11, the molecule of any one of claims 12-39, or the pharmaceutical composition of any one of claims 40-43, thereby treating the subject.

49. A method of cleaving B cell receptor, the method comprising administering a therapeutically effective amount of the immunoglobulin protease of any one of claims 1-11, the molecule of any one of claims 12-39, or the pharmaceutical composition of any one of claims 40-43, to cleave the B cell receptor.-254-IPTS / 200163498.2DOCKET NO. SES-020WO PATENT50. A nucleic acid encoding the immunoglobulin protease of any one of claims 1-11, the molecule of any one of claims 12-39, or the pharmaceutical composition of any one of claims 40-43.

51. A vector comprising the nucleic acid of claim 50.

52. A cell comprising the nucleic acid of claim 50 or the vector of claim 51.

53. A method of making the immunoglobulin protease of any one of claims 1-11, the molecule of any one of claims 12-39, comprising culturing a cell of claim 52 to make the immunoglobulin protease or the molecule.-255-IPTS / 200163498.2

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