Protease variants and uses thereof

Abstract

Embodiments provided herein provide polypeptides and molecules, including polypeptides and variant Fc molecules, with protease activity, pharmaceutical compositions comprising same, and methods that can be used to treat disorders, such as IgG-related disorders.

Description

[Technical Field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims priority to 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 / 493,450, filed March 31, 2023, U.S. Provisional Application No. 63 / 506,539, filed June 6, 2023, and U.S. Provisional Application No. 63 / 600,157, filed November 17, 2023, each of which is incorporated by reference in its entirety.

[0002] Reference to an electronically submitted sequence listing This application contains a Sequence Listing that has been submitted electronically in XML file format, which is incorporated herein by reference in its entirety. The XML copy, created on January 4, 2024, is named "SES-007WO_SEQ.XML" and is 1,635,203 bytes in size.

[0003] Embodiments provided herein relate to polypeptides having protease activity and polypeptides comprising an Fc portion, and compositions comprising the same. [Background technology]

[0004] Streptococcus pyogenes immunoglobulin G-degrading enzyme (IdeS) is an extracellular cysteine ​​protease produced by the human pathogen Streptococcus pyogenes. IdeS has a very high degree of substrate specificity, with immunoglobulin G (IgG) being its only identified substrate. IdeS also catalyzes a single proteolytic cleavage in the lower hinge region of the heavy chains of all subclasses of human IgG. IdeS also catalyzes equivalent cleavage of the heavy chains of several subclasses of IgG in various animal species. IdeS efficiently cleaves IgG into Fc and F(ab')2 fragments. IdeS is a virulence factor of Streptococcus pyogenes, which causes common infections such as tonsillitis and streptococcal pharyngitis. To date, an IdeS-Fc molecule resistant to autocleavage has not been exploited. Pathogenic IgG antibodies constitute a significant clinical problem, contributing to the pathogenesis of numerous autoimmune conditions and acute transplant rejection. Therefore, the ability to effectively reduce or eliminate such antibodies is a significant clinical challenge. The embodiments provided herein meet this need as well as other needs. [Brief explanation of the drawings]

[0005] [Figure 1] 1 shows the performance of the IdeS-Fc polypeptide against IdeS. [Figure 2A] PK measurements are shown. [Figure 2B] PD measurements are shown. [Figure 3] PK and PD measurements are shown. [Figure 4A] The urine protein score is shown. [Figure 4B] The urine protein score is shown. [Figure 4C] Indicates blood urea nitrogen level. [Figure 4D] 1 shows an analysis of in vivo renal injury. [Figure 4E] 1 shows an analysis of in vivo renal injury. [Figure 4F] 1 shows an analysis of in vivo renal injury. [Figure 5A] 1 shows an analysis of B cell receptor kappa cleavage. [Figure 5B]1 shows an analysis of B cell receptor lambda cleavage. [Figure 6] 1 shows the results of a human plasma cleavage assay. [Figure 7A] 1 shows the results of a human plasma cleavage assay. [Figure 7B] 1 shows the results of a human plasma cleavage assay. [Figure 8] Figure 1 shows reduced levels of preformed IgG binding to the protease mutant compared to wild-type Fc-IdeS. [Figure 9] 1 shows an alignment of non-limiting protease variants to the wild-type protease sequence. DETAILED DESCRIPTION OF THE INVENTION

[0006] As used in this specification and the appended claims, the singular forms "a," "an," and "the" include plural referents unless the context clearly dictates otherwise.

[0007] As used herein, the term "about" means that a numerical value is approximate and that small variations do not significantly affect the practice of the disclosed embodiments. When a numerical limit is used, unless otherwise indicated by context, "about" means that the numerical value can vary by ±5% and remain within the range of the disclosed embodiments. Thus, about 100 means 95 to 105.

[0008] 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., mouse, rat, or guinea pig), monkey, cat, dog, cow, horse, pig, or human. In some embodiments, the mammal is a human.

[0009] As used herein, the term "contacting" refers to bringing two elements together in an in vitro system or in an in vivo system. For example, "contacting" a therapeutic compound with an individual or patient or cell includes administering the compound to an individual or patient, such as a human, and introducing the compound into a sample containing, for example, a cell or purified preparation containing the target.

[0010] As used herein, the terms "comprising" (and any form of comprising, e.g., "comprise," "comprises," and "comprised"), "having" (and any form of having, e.g., "have" and "has"), "including" (and any form of including, e.g., "includes" and "include"), or "containing" (and any form of containing, e.g., "contains" and "contain") are inclusive or open-ended and do not exclude additional, unrecited, elements or method steps. Any composition or method reciting the term "comprising" should be understood to also describe a composition that consists of, consists of, or consists essentially of the recited components or elements.

[0011] As used herein, the terms "fused" or "linked," when used in reference to proteins or molecules having different domains or heterologous sequences, mean that the protein domains are part of the same peptide chain, connected to each other either by peptide bonds or other covalent bonds. The domains or segments can be directly linked or fused to each other, or another domain or peptide sequence can be between the two domains or sequences, and such sequences would still be considered fused or linked to each other.

[0012] As used herein, the terms "individual," "subject," or "patient," used interchangeably, refer to any animal, including a mammal, e.g., a mouse, rat, other rodent, rabbit, dog, cat, pig, cow, sheep, horse, or primate, such as a human.

[0013] As used herein, the term "inhibit" refers to a result, symptom, or activity that is reduced compared to the activity or result in the absence of the compound 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%. Consequently, a symptom or activity can also be inhibited if it is completely eliminated or abolished.

[0014] As used herein, the phrase "in need of" means that a subject has been identified as having a need for a particular method or treatment. In some embodiments, identification can be by any means of diagnosis. A subject may be in need of any of the methods and treatments described herein. In some embodiments, the subject is in or moves to an environment where a particular disease, disorder, or condition is prevalent.

[0015] As used herein, the phrase "an integer from X to Y" means any integer, including the endpoints. For example, the phrase "an integer from 1 to 5" means 1, 2, 3, 4, or 5.

[0016] As used herein, the phrase "ophthalmically acceptable" means having no lasting adverse effects on the treated eye or its function, or on the general health of the treated subject. However, it is recognized that transient effects, such as mild irritation or a "stinging" sensation, are common with topical ophthalmic administration of drugs, and the presence of such transient effects is not inconsistent with the composition, formulation, or ingredients (e.g., excipients) in question being "ophthalmically acceptable," as defined herein. In some embodiments, a pharmaceutical composition may be ophthalmically acceptable or suitable for ophthalmic administration.

[0017] In some embodiments, the term "therapeutic molecule" can be used interchangeably with "therapeutic compound," "molecule," or "therapeutic agent," and refers to any polypeptide or protein described herein.

[0018] As used herein, the term "position" refers to a position in a polypeptide sequence. Positions may be numbered consecutively or according to established formats, such as the EU numbering system based on Kabat amino acid positions. For example, position 298 is a position in human antibody IgG1.

[0019] "Specific binding" or "specifically binds to" or "specific for" a particular antigen, target, or epitope means binding that is measurably different from non-specific interactions. Specific binding can be measured, for example, by determining the binding of a molecule compared to the binding of a control molecule, which is generally 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.

[0020] Specific binding to a particular antigen, target, or epitope can be measured, for example, by measuring the K D is at least about 10 -4M , at least about 10 -5M , at least about 10 -6M , at least about 10-7M , at least about 10 -8M , at least about 10 -9M , alternatively at least about 10 -10M , at least about 10 -11M , at least about 10 -12M or more, where K D refers to the off-rate of a particular antibody-target interaction. Typically, an antibody that specifically binds to an antigen or target has a K that is 2-fold, 4-fold, 5-fold, 10-fold, 20-fold, 50-fold, 100-fold, 500-fold, 1000-fold, 5,000-fold, 10,000-fold, or more greater than a control molecule, or at least 2-fold, 4-fold, 5-fold, 10-fold, 20-fold, 50-fold, 100-fold, 500-fold, 1000-fold, 5,000-fold, 10,000-fold, or more greater than a control molecule, compared to the antigen or epitope. D It has.

[0021] In some embodiments, specific binding to a particular antigen, target, or epitope is, for example, a K for the target, antigen, or epitope that is at least 2-fold, 4-fold, 5-fold, 20-fold, 50-fold, 100-fold, 500-fold, 1000-fold, 5,000-fold, 10,000-fold, or more, greater than a control. A or K a where K A or K a refers to the association rate of a particular antibody-antigen interaction.

[0022] As provided herein, the compounds and compositions provided herein can be used in the therapeutic methods provided herein. As used herein, the terms "treat," "treated," or "treating" refer to both therapeutic treatments and prophylactic measures, the purpose of which is to slow (alleviate) an undesirable physiological condition, disorder, or disease, or to obtain a beneficial or desired clinical result. For purposes of these embodiments, a beneficial or desired clinical result includes, but is not limited to, alleviation of symptoms, a decrease in the extent of a condition, disorder, or disease, a stabilized (i.e., not worsening) condition, disorder, or disease, a delay or slowing of the progression of a condition, disorder, or disease, an improvement or remission of a condition, disorder, or disease state, whether detectable or undetectable, an improvement in at least one measurable physical parameter, not necessarily discernible by the patient, or an enhancement or amelioration of a condition, disorder, or disease. Treatment includes eliciting a clinically significant response without excessive levels of side effects. Treatment, where applicable to a particular disease, also includes prolonging survival compared to expected survival if not receiving treatment. Thus, "treatment of an autoimmune condition" or "treating autoimmunity" refers to the activity of alleviating or ameliorating any of the primary phenomena or secondary symptoms associated with the autoimmune conditions described herein, as well as other conditions, when the terms "treat," "treated," or "treating" are used in conjunction with such conditions.

[0023] As used herein, the terms "variant," "molecule," "therapeutic agent," "therapeutic compound," "compound," "polypeptide," or "protein" can be used interchangeably and refer to the variants, molecules, therapeutic agents, therapeutic compounds, compounds, polypeptides, and proteins disclosed herein.

[0024] Variant Fc molecules As used herein, "isotype" refers to the immunoglobulin class (e.g., IgG1, IgG2, IgG3, IgG4, IgM, IgA1, IgA2, IgD, and IgE antibodies) encoded by heavy chain constant domain genes. The full-length amino acid sequence of each wild-type human IgG constant region (including all domains, i.e., CH1 domain, hinge, CH2 domain, and CH3 domain) is cataloged in the UniProt database available online, e.g., as P01857 (IgG1), P01859 (IgG2), P01860 (IgG3), and P01861 (IgG4), or their different allotypes (SEQ ID NOs: 1, 2, 3, and 4, respectively). As used herein, a domain of a heavy chain constant region, e.g., a hinge, is of the "IgG1 isotype," "IgG2 isotype," "IgG3 isotype," or "IgG4 isotype" if the domain comprises the amino acid sequence of the corresponding domain of each isotype or a variant thereof (which has higher homology to the corresponding domain of each isotype than to that of other isotypes).

[0025] "Allotype" refers to naturally occurring variants within a particular isotype group, which variants differ in some amino acids (see, e.g., Jefferies et al. (2009) mAbs 1:1). The molecules described herein can be of any allotype.

[0026] A "wild-type" protein or portion thereof is a version of a protein found in nature. The amino acid sequence of a wild-type protein, e.g., a heavy chain constant region, is the amino acid sequence of a naturally occurring protein. Due to allotypic differences, there may be more than one amino acid sequence for a wild-type protein. For example, there are several allotypes of the naturally occurring human IGg1 heavy chain constant region (see, e.g., Jeffries et al. (2009) mAbs 1:1).

[0027] As used herein, the term "chemical liability" is meant to refer to factors that affect the immunogenicity of a molecule. Thus, in some embodiments, "chemical liability" is meant to refer to, but is not limited to, post-translational modifications, aggregation, glycosylation, impurities, or formulation components.

[0028] Immunoglobulins can be from any of the commonly known isotypes, including, but not limited to, IgA, secretory IgA, IgG, and IgM. IgG isotypes are divided into subclasses in certain species, such as IgG1, IgG2, IgG3, and IgG4 in humans and IgG1, IgG2a, IgG2b, and IgG3 in mice. In certain embodiments, the antibodies described herein are of the human IgG1 or IgG2 subtype. Immunoglobulins, such as human IgG1, exist in several allotypes that differ from each other in at most a few amino acids.

[0029] In some embodiments, the IgG protein (hinge region underlined) is as provided in Table 1. [Table 1]

[0030] "Fc region" (Fragment Crystallizable Region) or "Fc domain" or "Fc" refers to the C-terminal region of an antibody heavy chain that mediates binding of immunoglobulins 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 (C1q) of the classical complement system. Thus, the Fc region of an antibody of isotype IgG comprises the heavy chain constant region of the antibody excluding the first constant region immunoglobulin domain (CH1). 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, while IgM and IgE Fc regions comprise three heavy chain constant domains (CH domains 2-4) in each polypeptide chain. In the case of IgG, the Fc region comprises immunoglobulin domains consisting of the hinge, CH2, and CH3. For purposes herein, the Fc region is defined as beginning at amino acid 216 and ending at amino acid 447, with numbering according to the EU index as per Kabat (Kabat et al. (1991) Sequences of Proteins of Immunological Interest, National Institutes of Health, Bethesda, MD) and according to Figures 3c-3f of U.S. Patent Application Publication No. 2008 / 0248028. In some embodiments, the Fc region includes the hinge region. The Fc can be a native (or naturally occurring or wild-type) Fc, including any allotypic variants, or a variant Fc (e.g., a non-naturally occurring Fc), including, for example, 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 wild-type Fc. A modified or mutated Fc may have enhanced or reduced effector function and / or half-life. Fc may refer to this region in isolation or in the context of a protein polypeptide that comprises the Fc, such as an "Fc region-containing binding protein" (e.g., an antibody or immunoadhesin), also referred to as an "Fc fusion protein."In some embodiments, the modified or variant Fc molecule has enhanced binding to FcγRIIb.

[0031] The terms "hinge," "hinge domain," or "hinge region," or "antibody hinge region" refer to the domain of the heavy chain constant region that connects the CH1 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 a site for intermolecular disulfide bonds between the two heavy chain constant regions. The term "hinge" includes wild-type hinges (such as those listed in Table 3) as well as variants thereof (e.g., non-naturally occurring hinges or modified hinges). For example, the term "IgG1 hinge" includes the wild-type IgG1 hinge shown below as well as variants having 1, 2, 3, 4, 5, 1-3, 1-5, 3-5, and / or up to 5, 4, 3, 2, or 1 mutations, e.g., substitutions, deletions, or additions. In some embodiments, the hinge region is as provided in Table 2. [Table 2]

[0032] The term "CH1 domain" refers to a heavy chain constant region that connects a variable domain to a hinge within a heavy chain constant domain. As used herein, CH1 domains include wild-type CH1 domains and variants thereof (e.g., non-naturally occurring CH1 domains or modified CH1 domains). For example, the term "CH1 domain" includes wild-type CH1 domains and variants thereof having 1, 2, 3, 4, 5, 1-3, 1-5, 3-5, and / or up to 5, 4, 3, 2, or 1 mutations, e.g., substitutions, deletions, or additions.

[0033] The term "CH2 domain" refers to a heavy chain constant region that connects the hinge to the CH3 domain within the heavy chain constant domain. As used herein, CH2 domains include wild-type CH2 domains and 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 up to 5, 4, 3, 2, or 1 mutations, e.g., substitutions, deletions, or additions.

[0034] The term "CH3 domain" refers to a heavy chain constant region that is C-terminal to the CH2 domain in a heavy chain constant domain. As used herein, CH3 domains include wild-type CH3 domains and 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 up to 5, 4, 3, 2, or 1 mutations, e.g., substitutions, deletions, or additions.

[0035] Provided herein are mutant Fc polypeptides, including mutant Fc polypeptides, such as Fc polypeptides having mutated sequence regions relative to a wild-type Fc polypeptide. Exemplary mutant Fc molecules, including mutant Fc polypeptides, include an IgG1 hinge, CH1 domain, CH2 domain, and CH3 domain, where at least one of these constant domains has a residue that is not the wild-type residue compared to SEQ ID NO: 1, 2, 3, or 4. The mutant Fc molecules can have effector functions similar to those of wild-type IgG, or can be engineered to have enhanced effector functions relative to those of wild-type IgG. The variant Fc molecule may comprise a wild-type CH1, hinge, CH2 and / or CH3 domain, or a variant thereof, e.g., a CH1, hinge, CH2 and / or CH3 domain having one or more amino acid substitutions, deletions, or additions compared 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.

[0036] In some embodiments, the variant Fc polypeptide comprises one or more mutations that confer resistance to protease recognition. In some embodiments, the variant Fc polypeptide comprises one or more mutations that confer resistance to proteolytic cleavage. In some embodiments, the variant Fc polypeptide comprises one or more mutations that confer resistance to protease recognition and proteolytic cleavage.

[0037] In some embodiments, the 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 an amino acid mutation at any one or more positions from 234 to 329 compared to SEQ ID NO:1, wherein the mutation comprises an insertion, deletion, or substitution.

[0038] In some embodiments, the 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 mutation corresponding to the L234A mutation compared to SEQ ID NO: 1.

[0039] In some embodiments, the 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 mutation corresponding to the L235A mutation compared to SEQ ID NO: 1.

[0040] In some embodiments, the 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 mutation corresponding to a G237A mutation compared to SEQ ID NO:1.

[0041] In some embodiments, the 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 mutation corresponding to the Y296Q mutation compared to SEQ ID NO:1.

[0042] In some embodiments, the 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 mutation corresponding to the P329K mutation compared to SEQ ID NO: 1.

[0043] In some embodiments, the 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 at least 100% sequence identity to SEQ ID NO: 1, provided that the variant Fc polypeptide comprises a set of mutations corresponding to the L234A, L235A, and G237A mutations compared to SEQ ID NO: 1.

[0044] In some embodiments, the 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 the Y296Q and P329K mutations compared to SEQ ID NO: 1.

[0045] In some embodiments, the 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 the L234A, L235A, G237A, Y296Q, and P329K mutations compared to SEQ ID NO:1.

[0046] In some embodiments, the variant Fc polypeptide comprises the set of mutations L234A, L235A, G237A, Y296Q, and P329K compared to SEQ ID NO:1, and the variant Fc polypeptide further comprises at least, about, or exactly 1 to 10, 1 to 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 set of mutations compared to SEQ ID NO:1. [Table 3-1] [Table 3-2]

[0047] In some embodiments, the 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 any of the sequences provided in Table 3. In some embodiments, the 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, 25, 42, 43, 44, 45, 46, 47, 48, or 49. In some embodiments, the 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.

[0048] 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 any of the sequences provided in Table 3, provided that the variant Fc polypeptide comprises one or more mutations at position 234, 235, 237, 265, 269, 296, 298, 329, or any combination thereof. In some embodiments, the 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 one or more mutations at position 234, 235, 237, 265, 269, 296, 298, 329, or any combination thereof. The 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 one or more mutations at positions 234, 235, 237, 265, 269, 296, 298, 329, or any combination thereof.

[0049] In some embodiments, the 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 any of the sequences provided in Table 3, and comprises one or more mutations: L234A, L235A, G237A, Y296Q, P329K, or any combination thereof. In some embodiments, the 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 one or more mutations: L234A, L235A, G237A, Y296Q, P329K, or any combination thereof. In some embodiments, the 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 one or more mutations: L234A, L235A, G237A, Y296Q, P329K, or any combination thereof.

[0050] In some embodiments, the 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 the L234A, L235A, G237A, Y296Q, and P329K mutations. In some embodiments, the 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 the L234A, L235A, G237A, Y296Q, and P329K mutations.

[0051] In some embodiments, the variant Fc polypeptide comprises the set of mutations L234A, L235A, G237A, Y296Q, and P329K compared to SEQ ID NO: 19, 20, 21, 22, 23, 24, or 25, and the variant Fc polypeptide further comprises at least, about, or exactly 1 to 10, 1 to 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 set of mutations L234A, L235A, G237A, Y296Q, and P329K compared to SEQ ID NO: 19, 20, 21, 22, 23, 24, or 25. In some embodiments, the variant Fc polypeptide comprises the set of mutations L234A, L235A, G237A, Y296Q, and P329K compared to SEQ ID NO:23, and the variant Fc polypeptide further comprises at least, about, or exactly 1 to 10, 1 to 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 set of mutations L234A, L235A, G237A, Y296Q, and P329K compared to SEQ ID NO:23.

[0052] In some embodiments, a variant Fc polypeptide comprises the amino acid sequence provided in Table 3. In some embodiments, a variant Fc polypeptide comprises the amino acid sequence of SEQ ID NO: 19, 20, 21, 22, 23, 24, 25, 42, 43, 44, 45, 46, 47, 48, or 49. In some embodiments, a variant Fc polypeptide comprises the amino acid sequence of SEQ ID NO: 19. In some embodiments, a variant Fc polypeptide comprises the amino acid sequence of SEQ ID NO: 20. In some embodiments, a variant Fc polypeptide comprises the amino acid sequence of SEQ ID NO: 21. In some embodiments, a variant Fc polypeptide comprises the amino acid sequence of SEQ ID NO: 22. In some embodiments, a variant Fc polypeptide comprises the amino acid sequence of SEQ ID NO: 23. In some embodiments, a variant Fc polypeptide comprises the amino acid sequence of SEQ ID NO: 24. In some embodiments, a variant Fc polypeptide comprises the amino acid sequence of SEQ ID NO: 25. In some embodiments, a variant Fc polypeptide comprises the amino acid sequence of SEQ ID NO: 42. In some embodiments, a variant Fc polypeptide comprises the amino acid sequence of SEQ ID NO: 43. In some embodiments, a variant Fc polypeptide comprises the amino acid sequence of SEQ ID NO: 44. In some embodiments, a variant Fc polypeptide comprises the amino acid sequence of SEQ ID NO: 45. In some embodiments, a variant Fc polypeptide comprises the amino acid sequence of SEQ ID NO: 46. In some embodiments, a variant Fc polypeptide comprises the amino acid sequence of SEQ ID NO: 47. In some embodiments, a variant Fc polypeptide comprises the amino acid sequence of SEQ ID NO: 48. In some embodiments, a variant Fc polypeptide comprises the amino acid sequence of SEQ ID NO: 49.

[0053] In some embodiments, the variant Fc polypeptide comprises an amino acid sequence that has a C-terminal lysine (K). In some embodiments, the variant Fc polypeptide comprises an amino acid sequence that does not have a C-terminal lysine (K).

[0054] In some embodiments, a variant Fc polypeptide such as those provided herein is conjugated to another polypeptide. In some embodiments, the other polypeptide is an antibody. In some embodiments, a variant Fc polypeptide such as those provided herein is conjugated to an effector binding / modulating polypeptide or a tissue-targeting polypeptide. In some embodiments, a variant Fc polypeptide such as those provided herein is conjugated to an antibody, or an antigen-binding fragment thereof. In some embodiments, such a variant Fc polypeptide is conjugated to another polypeptide via a linker or covalent bond. In some embodiments, the linker is a protein linker such as those provided herein.

[0055] Protease variants The present disclosure provides polypeptides, molecules, compounds, therapeutic agents, and compositions comprising a polypeptide with protease activity, such as those provided herein, that can be covalently or non-covalently linked to an Fc polypeptide domain. In some embodiments, the polypeptide with protease activity is IdeS, IdeSsuis, IdeZ, IdeE, IdeE2, IdeZ2, or IdeC.

[0056] Streptococcus pyogenes is an important bacterial pathogen that secretes two enzymes, EndoS and IdeS, that show remarkable specificity for IgG. EndoS, an endoglycosidase of S. pyogenes, specifically hydrolyzes functionally important N-linked glycans on IgG, and treatment with EndoS abolishes 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 from Streptococcus pyogenes) is a cysteine ​​protease that cleaves IgG with unique specificity in the hinge region. (Wenig K, Chatwell L, von Pawel-Rammingen U, Bjoerck 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 highly specific for IgG, hydrolyzing both heavy chains at the hinge region after glycine residue 236 to generate one F(ab')2 and two monomeric Fc fragments. IdeE is a homolog of the secreted IgG-specific protease IdeS / Mac from Streptococcus pyogenes. The activity of IdeE is comparable to that of the corresponding enzyme IdeZ from the closely related Streptococcus equi species zooepidemicus.

[0057] The full sequence of IdeS is publicly available under NCBI reference sequence number WP_010922160.1 and is provided herein as SEQ ID NO: 35: MRKRCYSTSAAVLAAVTLFVLSVDRGVIADSFSANQEIRYSEVTPYHVTSVWTKGVTPPANFTQGEDVFHAPYVANQGWYDITKTFNGKDDLLCGAATAGNMLHWWFDQNKDQIKRYLEEHPEKQKINFNGEQMFDVKEAIDTKNHQLDSKLFEYFKEKAFPYLSTKHLGVFPDHVIDMFINGYRLSLTNHGPTPVKEGSKDPRGGIFDAVFTRGDQSKLLTSRHDFKEKNLKEISDLIKKELTEGKALGLSHTYANVRINHVINLWGADFDSNGNLKAIYVTDSDSNASIGMKKYFVGVNSAGKVAISAKEIKEDNIGAQVLGLFTLSTGQDSWNQTN (SEQ ID NO: 35).

[0058] This sequence includes an N-terminal methionine followed by a 28-amino acid secretion signal sequence. The N-terminal methionine and signal sequence (totaling 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 under UniProt identifier Q9F1R7_STRPY and is provided herein as SEQ ID NO: 36. Another variant of IdeS protease that can be used as a reference sequence may be as set forth in SEQ ID NO: 1349, which is similar to the amino acid sequence of SEQ ID NO: 36, but has four additional amino acid residues at the N-terminus that are DSFS (SEQ ID NO: 1350).

[0059] IdeZ is an IgG cysteine ​​protease produced by Streptococcus equi species zooepidemicus, a bacterium found primarily in horses. Because IdeZ is not a human pathogen, human subjects typically do not have antibodies to this protein in their plasma. However, IdeZ has a level of IgG cysteine ​​protease activity against human IgG that is significantly lower than that of IdeS. The full sequence of IdeZ is publicly available under the UniProt identifier Q0PIW1 and is provided herein as SEQ ID NO: 1331. MKTIAYPNKPHSLSAGLLTAIAIFSLASSNITYADDYQRNAAEVYAKEVPHQITSVWTKGVTPLTPEQFRYNNEDVIHAPYLAHQGWYDITKVFDGKDNLLCGAATAGNMLHWWFDQNKTEIEAYLSKHPEKQKIIFNNQELFDLKAAIDTKDSQTNSQLFNYFRDKAFPNLSARQLGVMPDLVLDMFINGYYLNVFKTQSTDVNRPYQDKDKRGGIFDAVFTRGDQTTLLTARHDLKNKGLNDISTIIKQELTEGRALALSHTYANVSISHVINLWGADFNAEGNLEAIYVTDSDANASIGMKKYFVGINAHGHVAISAKKIEGENIGAQVLGLFTLSSGKDIWQKLS (SEQ ID NO: 1331).

[0060] In some embodiments, IdeZ is publicly available under NCBI reference sequence number WP_014622780.1 and is provided herein as SEQ ID NO: 1332 below: MKTIAYPNKPHSLSAGLLTAIAIFSLASSNITYADDYQRNATEAYAKEVPHQITSVWTKGVIPLIPEQFRYNNEDVIHAPYLAHQGWYDITKAFDGKDNLLCGAATAGNMLHWWFDQNKTEIEAYLSKHPEKQKIIFNNQELFDLKAAIDTKDSQINSQLFNYFRDKAFPNLSARQLGVMPDLVLDMFINGYYLNVFKIQSTDVNRPYQDKDKRGGIFDAVFIRGDQTILLTARHDLKNKGLNDISTIIKQELTEGRALALSHIYANVSISHVINLWGADFNAEGNLEAIYVIDSDANASIGMKKYFVGINAHGHVAISAKKIEGENIGAQVLGLFTLSSGKDIWQKLS (SEQ ID NO: 1332).

[0061] The mature sequence of IdeZ is publicly available under the UniProt identifier A0A0D0YKS5_STRSZ and is provided herein as SEQ ID NO: 38.

[0062] 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 under UniProt identifier C5W022 and is provided herein as SEQ ID NO: 55. The mature sequence of IdeE is publicly available under UniProt identifier C0M8U6_STRE4 and is provided herein as SEQ ID NO: 37. The mature sequence of IdeE2 is publicly available under UniProt identifier C7B615_9STRE and is provided herein as SEQ ID NO: 39. The mature sequence of IdeZ2 is publicly available under UniProt identifier B4U2F7_STREM and is provided herein as SEQ ID NO: 40. The mature sequence of IdeC is publicly available under the 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. [Table 4-1] [Table 4-2] [Table 4-3]

[0063] In some embodiments, the 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 of the sequences 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, the 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.

[0064] In some embodiments, the polypeptide having protease activity comprises an amino acid 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. 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, the polypeptide having protease activity comprises the amino acid sequence of SEQ ID NO: 26, 28, 29, 31, 36, 37, 38, 39, 40, 41, 50, 51, 52, or 55.

[0065] In some embodiments, the protease comprises an amino acid sequence that includes a signal sequence. In some embodiments, the protease comprises an amino acid sequence that does not include a signal sequence. In some embodiments, the signal sequence can have the amino acid sequence of METDTLLLWVLLLWVPGSTG (SEQ ID NO: 53), or MNIQERFSLRKSAVGLVSVSLLCAIYTSTVAA (SEQ ID NO: 54).

[0066] In some embodiments, the protease has a sequence selected from Table 5. [Table 5-1] [Table 5-2] [Table 5-3] [Table 5-4] Table 5-5 Table 5-6 Table 5-7 Table 5-8 Table 5-9 Table 5-10 Table 5-11 Table 5-12 Table 5-13 Table 5-14 Table 5-15 Table 5-16 Table 5-17 Table 5-18 Table 5-19 Table 5-20 Table 5-21 Table 5-22 Table 5-23 Table 5-24 Table 5-25 Table 5-26 Table 5-27 Table 5-28 Table 5-29 Table 5-30 Table 5-31 Table 5-32 Table 5-33 Table 5-34 Table 5-35 Table 5-36 Table 5-37 Table 5-38 Table 5-39 Table 5-40 Table 5-41 Table 5-42 Table 5-43 Table 5-44 Table 5-45 Table 5-46 Table 5-47 Table 5-48 Table 5-49 Table 5-50 Table 5-51 Table 5-52 Table 5-53 Table 5-54 Table 5-55 Table 5-56 Table 5-57 Table 5-58 Table 5-59 Table 5-60 Table 5-61 Table 5-62 Table 5-63 Table 5-64 Table 5-65 Table 5-66 Table 5-67 Table 5-68 Table 5-69 Table 5-70 Table 5-71 Table 5-72 Table 5-73 Table 5-74 Table 5-75 Table 5-76 Table 5-77 Table 5-78 Table 5-79 Table 5-80 Table 5-81

[0067] 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%, 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 of the amino acid sequences selected from SEQ ID NOs: 55-1019, 1290-1310, and 1333. A protease 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 of the amino acid sequences selected from SEQ ID NOs: 55 to 1019, 1290 to 1310, and 1333. These and other related applications may be found in 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 incorporated herein by reference in its entirety.

[0068] 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%, 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: 1290. 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%, 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: 1291. 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%, 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: 1292. 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%, 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: 1293. 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%, 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:1294.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%, 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: 1295. 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%, 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: 1296. 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%, 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: 1297. 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%, 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: 1298. 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%, 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:1299.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%, 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: 1300. 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%, 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: 1301. 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%, 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: 1302. 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%, 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: 1303. 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%, 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: 1304.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%, 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: 1305. 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%, 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: 1306. 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%, 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: 1307. 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%, 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: 1308. 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%, 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: 1309.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%, 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: 1310. 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%, 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: 1333.

[0069] 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%, 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 any one of SEQ ID NOs: 56-752, and optionally does not include a methionine (M) residue at the N-terminus of any one of SEQ ID NOs: 56-752. For example, a protease 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 the amino acid sequence of any one of SEQ ID NOs: 56-752, and optionally not containing a methionine (M) residue at the N-terminus of any one of SEQ ID NOs: 56-752, may have at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, The amino acid sequence has 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, and the amino acid sequence has 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 any one of SEQ ID NOs: 56 to 752, and does not contain a methionine (M) residue at the N-terminus.

[0070] In some embodiments, the protease has an amino acid sequence selected from SEQ ID NOs: 55-1019, 1290-1310, and 1333. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 56. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 57. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 58. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 59. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 60. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 61. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 62. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 63. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 64. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 65. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 66. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 67. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 68. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 69. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 70. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 71. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 72. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 73. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 74. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 75. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 76. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 77. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 78. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 79.In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 80. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 81. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 82. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 83. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 84. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 85. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 86. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 87. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 88. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 89. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 90. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 91. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 92. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 93. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 94. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 95. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 96. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 97. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 98. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 99. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 100. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 101. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 102. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 103. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 104. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 105.In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 106. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 107. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 108. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 109. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 110. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 111. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 112. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 113. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 114. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 115. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 116. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 117. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 118. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 119. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 120. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 121. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 122. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 123. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 124. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 125. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 126. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 127. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 128. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 129. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 130.In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 131. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 132. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 133. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 134. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 135. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 136. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 137. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 138. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 139. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 140. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 141. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 142. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 143. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 144. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 145. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 146. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 147. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 148. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 149. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 150. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 151. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 152. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 153. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 154. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 155.In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 156. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 157. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 158. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 159. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 160. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 161. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 162. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 163. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 164. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 165. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 166. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 167. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 168. In some embodiments, the protease has the amino acid sequence of SEQ ID NO:169. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 170. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 171. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 172. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 173. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 174. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 175. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 176. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 177. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 178. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 179. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 180. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 181. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 182. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 183. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 184. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 185. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 186. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 187. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 188. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 189. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 190. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 191. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 192. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 193. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 194.In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 195. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 196. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 197. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 198. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 199. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 200. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 201. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 202. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 203. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 204. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 205. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 206. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 207. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 208. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 209. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 210. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 211. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 212. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 213. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 214. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 215. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 216. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 217. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 218. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 219.In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 220. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 221. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 222. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 223. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 224. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 225. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 226. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 227. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 228. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 229. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 230. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 231. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 232. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 233. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 234. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 235. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 236. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 237. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 238. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 239. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 240. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 241. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 242. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 243. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 244.In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 245. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 246. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 247. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 248. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 249. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 250. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 251. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 252. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 253. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 254. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 255. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 256. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 257. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 258. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 259. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 260. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 261. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 262. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 263. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 264. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 265. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 266. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 267. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 268. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 269.In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 270. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 271. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 272. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 273. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 274. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 275. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 276. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 277. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 278. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 279. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 280. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 281. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 282. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 283. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 284. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 285. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 286. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 287. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 288. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 289. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 290. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 291. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 292. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 293. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 294.In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 295. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 296. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 297. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 298. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 299. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 300. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 301. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 302. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 303. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 304. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 305. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 306. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 307. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 308. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 309. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 310. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 311. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 312. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 313. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 314. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 315. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 316. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 317. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 318. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 319. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 320. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 321. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 322. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 323. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 324.In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 325. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 326. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 327. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 328. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 329. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 330. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 331. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 332. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 333. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 334. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 335. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 336. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 337. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 338. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 339. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 340. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 341. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 342. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 343. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 344. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 345. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 346. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 347. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 348. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 349.In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 350. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 351. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 352. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 353. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 354. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 355. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 356. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 357. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 358. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 359. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 360. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 361. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 362. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 363. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 364. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 365. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 366. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 367. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 368. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 369. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 370. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 371. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 372. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 373. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 374.In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 375. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 376. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 377. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 378. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 379. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 380. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 381. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 382. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 383. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 384. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 385. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 386. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 387. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 388. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 389. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 390. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 391. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 392. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 393. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 394. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 395. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 396. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 397. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 398. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 399. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 400. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 401. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 402. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 403. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 404. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 405. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 406. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 407. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 408. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 409. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 410. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 411. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 412. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 413. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 414. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 415. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 416. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 417. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 418. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 419. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 420. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 421. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 422. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 423. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 424.In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 425. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 426. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 427. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 428. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 429. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 430. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 431. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 432. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 433. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 434. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 435. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 436. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 437. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 438. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 439. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 440. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 441. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 442. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 443. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 444. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 445. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 446. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 447. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 448. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 449.In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 450. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 451. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 452. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 453. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 454. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 455. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 456. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 457. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 458. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 459. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 460. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 461. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 462. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 463. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 464. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 465. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 466. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 467. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 468. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 469. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 470. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 471. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 472. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 473. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 474.In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 475. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 476. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 477. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 478. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 479. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 480. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 481. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 482. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 483. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 484. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 485. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 486. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 487. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 488. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 489. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 490. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 491. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 492. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 493. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 494. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 495. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 496. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 497. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 498. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 499. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 500. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 501. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 502. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 503. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 504. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 505. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 506. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 507. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 508. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 509. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 510. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 511. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 512. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 513. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 514. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 515. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 516. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 517. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 518. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 519. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 520. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 521. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 522. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 523. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 524.In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 525. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 526. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 527. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 528. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 529. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 530. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 531. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 532. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 533. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 534. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 535. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 536. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 537. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 538. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 539. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 540. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 541. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 542. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 543. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 544. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 545. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 546. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 547. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 548. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 549.In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 550. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 551. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 552. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 553. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 554. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 555. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 556. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 557. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 558. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 559. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 560. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 561. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 562. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 563. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 564. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 565. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 566. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 567. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 568. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 569. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 570. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 571. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 572. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 573. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 574.In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 575. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 576. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 577. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 578. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 579. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 580. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 581. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 582. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 583. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 584. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 585. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 586. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 587. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 588. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 589. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 590. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 591. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 592. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 593. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 594. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 595. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 596. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 597. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 598. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 599. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 600. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 601. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 602. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 603. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 604. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 605. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 606. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 607. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 608. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 609. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 610. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 611. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 612. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 613. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 614. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 615. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 616. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 617. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 618. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 619. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 620. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 621. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 622. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 623. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 624.In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 625. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 626. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 627. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 628. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 629. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 630. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 631. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 632. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 633. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 634. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 635. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 636. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 637. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 638. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 639. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 640. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 641. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 642. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 643. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 644. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 645. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 646. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 647. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 648. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 649.In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 650. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 651. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 652. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 653. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 654. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 655. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 656. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 657. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 658. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 659. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 660. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 661. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 662. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 663. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 664. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 665. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 666. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 667. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 668. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 669. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 670. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 671. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 672. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 673. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 674.In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 675. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 676. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 677. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 678. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 679. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 680. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 681. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 682. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 683. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 684. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 685. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 686. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 687. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 688. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 689. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 690. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 691. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 692. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 693. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 694. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 695. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 696. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 697. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 698. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 699. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 700. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 701. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 702. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 703. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 704. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 705. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 706. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 707. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 708. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 709. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 710. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 711. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 712. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 713. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 714. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 715. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 716. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 717. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 718. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 719. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 720. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 721. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 722. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 723. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 724.In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 725. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 726. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 727. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 728. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 729. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 730. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 731. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 732. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 733. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 734. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 735. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 736. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 737. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 738. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 739. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 740. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 741. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 742. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 743. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 744. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 745. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 746. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 747. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 748. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 749.In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 750. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 751. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 752. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 753. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 754. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 755. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 756. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 757. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 758. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 759. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 760. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 761. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 762. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 763. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 764. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 765. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 766. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 767. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 768. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 769. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 770. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 771. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 772. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 773. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 774.In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 775. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 776. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 777. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 778. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 779. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 780. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 781. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 782. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 783. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 784. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 785. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 786. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 787. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 788. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 789. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 790. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 791. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 792. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 793. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 794. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 795. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 796. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 797. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 798. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 799. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 800. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 801. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 802. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 803. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 804. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 805. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 806. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 807. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 808. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 809. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 810. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 811. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 812. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 813. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 814. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 815. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 816. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 817. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 818. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 819. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 820. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 821. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 822. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 823. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 824.In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 825. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 826. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 827. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 828. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 829. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 830. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 831. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 832. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 833. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 834. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 835. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 836. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 837. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 838. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 839. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 840. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 841. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 842. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 843. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 844. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 845. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 846. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 847. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 848. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 849.In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 850. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 851. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 852. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 853. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 854. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 855. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 856. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 857. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 858. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 859. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 860. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 861. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 862. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 863. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 864. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 865. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 866. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 867. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 868. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 869. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 870. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 871. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 872. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 873. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 874.In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 875. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 876. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 877. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 878. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 879. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 880. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 881. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 882. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 883. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 884. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 885. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 886. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 887. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 888. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 889. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 890. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 891. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 892. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 893. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 894. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 895. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 896. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 897. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 898. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 899. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 900. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 901. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 902. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 903. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 904. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 905. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 906. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 907. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 908. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 909. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 910. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 911. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 912. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 913. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 914. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 915. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 916. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 917. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 918. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 919. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 920. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 921. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 922. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 923. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 924.In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 925. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 926. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 927. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 928. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 929. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 930. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 931. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 932. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 933. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 934. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 935. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 936. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 937. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 938. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 939. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 940. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 941. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 942. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 943. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 944. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 945. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 946. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 947. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 948. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 949.In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 950. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 951. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 952. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 953. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 954. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 955. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 956. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 957. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 958. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 959. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 960. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 961. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 962. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 963. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 964. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 965. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 966. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 967. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 968. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 969. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 970. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 971. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 972. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 973. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 974.In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 975. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 976. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 977. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 978. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 979. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 980. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 981. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 982. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 983. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 984. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 985. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 986. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 987. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 988. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 989. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 990. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 991. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 992. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 993. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 994. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 995. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 996. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 997. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 998. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 999. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 1000. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 1001. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 1002. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 1003. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 1004. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 1005. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 1006. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 1007. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 1008. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 1009. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 1010. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 1011. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 1012. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 1013. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 1014. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 1015. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 1016. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 1017. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 1018. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 1019. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 1290. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 1291. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 1292. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 1293. In some embodiments, the protease has the amino acid sequence of SEQ ID NO:1294.In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 1295. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 1296. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 1297. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 1298. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 1299. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 1300. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 1301. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 1302. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 1303. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 1304. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 1305. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 1306. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 1307. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 1308. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 1309. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 1310. In some embodiments, the protease has the amino acid sequence of SEQ ID NO: 1333.

[0071] In some embodiments, the protease has the amino acid sequence of any one of SEQ ID NOs: 56-752, and optionally does not include a methionine (M) residue at the N-terminus of any one of SEQ ID NOs: 56-752. For example, a protease having the amino acid sequence of any one of SEQ ID NOs: 56-752, and optionally does not include a methionine (M) residue at the N-terminus of any one of SEQ ID NOs: 56-752, comprises the amino acid sequence of any one of SEQ ID NOs: 56-752, and the amino acid sequence of any one of SEQ ID NOs: 56-752 does not include a methionine (M) residue at the N-terminus.

[0072] In some embodiments, the proteases and polypeptides provided herein comprise a leader sequence present at the N-terminus of the polypeptide. Without being bound by any particular theory, a leader sequence can facilitate expression of the polypeptide in eukaryotic cells. Any suitable leader sequence can be utilized. For example, a leader sequence comprising the sequence METDTLLLWVLLLWVPGSTG (SEQ ID NO: 1357) or MGWSCIILFLVATATGVHS (SEQ ID NO: 1358) can be used. These are non-limiting examples of leader sequences. The leader sequence is processed (removed) from the polypeptide as it is translated and expressed in a cell, leaving behind the polypeptide provided herein. Thus, in some embodiments, the polypeptides provided herein do not comprise a leader sequence when present in a pharmaceutical composition or as an isolated polypeptide. In some embodiments, the polypeptides provided 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 of the amino acid sequences selected from SEQ ID NOs: 55-1019, 1290-1310, and 1333, and further comprising 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%, 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 of the amino acid sequences selected from SEQ ID NOs: 55-1019, 1290-1310, and 1333, and further comprises a leader amino acid sequence having the amino acid sequence of METDTLLLWVLLLWVPGSTG (SEQ ID NO: 1357) or MGWSCIILFLVATATGVHS (SEQ ID NO: 1358).In some embodiments, the leader amino acid sequence is at the N-terminus of 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 ex vivo or in vivo expression.

[0073] In some embodiments, the protease comprises a polypeptide. In some embodiments, the polypeptide comprises a mutation at any position compared to SEQ ID NO: 35. In some embodiments, the mutation is a substitution, deletion, or insertion. In some embodiments, the mutation is a substitution. Non-limiting examples of positions that may contain a mutation compared to SEQ ID NO: 35 are provided in Table 20 below.

[0074] In some embodiments, the polypeptide comprises a mutation at any position provided in Table 21 relative to SEQ ID NO: 35. In some embodiments, the polypeptide comprises a set of mutations at any set of positions provided in Table 21 relative to SEQ ID NO: 35. In some embodiments, the polypeptide comprises an amino acid sequence having a mutation at any position provided in Table 21 relative to SEQ ID NO: 35. In some embodiments, the polypeptide comprises an amino acid sequence having a mutation at any set of positions provided in Table 21 relative to SEQ ID NO: 35. In some embodiments, the polypeptide has a sequence similar to SEQ ID NO:35, at least 50%, at least 51%, at least 52%, at least 53%, at least 54%, at least 55%, at least 56%, at least 57%, at least 58%, at least 59%, at least 60%, at least 61%, at least 62%, at least 63%, at least 64%, at least 65%, at least 66%, at least 67%, at least 68%, at least 69%, at least 70%, at least 71%, at least 72%, at least 73%, at least 74%, at least 75%, at least 76%, at least 77%, or at least 78% sequence similar to SEQ ID NO:35. Also includes an amino acid sequence having 78%, at least 79%, 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 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, provided that the amino acid sequence includes a mutation at any position, or any set of mutations at any set of positions, provided in Table 20, compared to SEQ ID NO: 35. [Table 6-1] [Table 6-2] [Table 6-3] Table 6-4 Table 6-5 Table 6-6 Table 6-7 Table 6-8 Table 6-9 Table 6-10 Table 6-11 Table 6-12 Table 6-13 Table 6-14 Table 6-15 Table 6-16 Table 6-17 Table 6-18 Table 6-19 Table 6-20 Table 6-21 Table 6-22 Table 6-23 Table 6-24 Table 6-25 Table 6-26 Table 6-27 Table 6-28 Table 6-29 Table 6-30 Table 6-31 Table 6-32 Table 6-33 Table 6-34 Table 6-35 Table 6-36 Table 6-37 [Table 6-38] [Table 6-39] [Table 6-40] [Table 6-41] [Table 6-42] [Table 6-43] [Table 6-44] [Table 6-45]

[0075] In some embodiments, the polypeptide comprises a mutation compared to SEQ ID NO: 35. In some embodiments, the mutation is a substitution, deletion, or insertion. In some embodiments, the mutation is a substitution. Non-limiting examples of mutations compared to SEQ ID NO: 35 are provided in Table 21 below.

[0076] In some embodiments, the polypeptide comprises any mutation provided in Table 21 relative to SEQ ID NO: 35. In some embodiments, the polypeptide comprises any set of mutations provided in Table 21 relative to SEQ ID NO: 35. In some embodiments, the polypeptide comprises an amino acid sequence having any set of mutations provided in Table 21 relative to SEQ ID NO: 35. In some embodiments, the polypeptide comprises an amino acid sequence having any set of mutations provided in Table 21 relative to SEQ ID NO: 35. In some embodiments, the polypeptide comprises an amino acid sequence having any set of mutations provided in Table 21 relative to SEQ ID NO: 35. In some embodiments, the polypeptide has a mutation that is at least 50%, at least 51%, at least 52%, at least 53%, at least 54%, at least 55%, at least 56%, at least 57%, at least 58%, at least 59%, at least 60%, at least 61%, at least 62%, at least 63%, at least 64%, at least 65%, at least 66%, at least 67%, at least 68%, at least 69%, at least 70%, at least 71%, at least 72%, at least 73%, at least 74%, at least 75%, at least 76%, at least or at least 97%, at least 78%, at least 79%, 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 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, provided that the amino acid sequence comprises any mutation or any set of mutations provided in Table 21 compared to SEQ ID NO: 35.

[0077] In some embodiments, the polypeptide comprises 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% sequence identity to SEQ ID NO: 35, However, the polypeptide comprises a mutation or set of mutations at any one position or any combination of positions selected from 60, 61, 63, 64, 69, 74, 76, 85, 112, 115, 116, 119, 120, 121, 130, 134, 145, 146, 154, 190, 198, 223, 225, 227, 231, 241, 251, 258, 261, 273, 274, 278, 286, 289, 303, 314, 334, and 336 relative to SEQ ID NO: 35. In some embodiments, the polypeptide has 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% sequence identity to SEQ ID NO: 1290, 1295, 1301, 1303, or 1310. and 336, except that the polypeptide comprises a mutation or set of mutations at any one position or any combination of positions selected from 60, 61, 63, 64, 69, 74, 76, 85, 112, 115, 116, 119, 120, 121, 130, 134, 145, 146, 154, 190, 198, 223, 225, 227, 231, 241, 251, 258, 261, 273, 274, 278, 286, 289, 303, 314, 334, and 336 relative to SEQ ID NO: 35.In some embodiments, the polypeptide has a sequence similar to that of SEQ ID NO: 1290, 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%. % sequence identity to SEQ ID NO: 35, except that the polypeptide comprises a set of mutations at positions 60, 61, 63, 64, 69, 74, 76, 85, 112, 115, 116, 119, 120, 121, 130, 134, 145, 146, 154, 190, 198, 223, 225, 227, 231, 241, 251, 273, 274, 278, 289, 303, 314, 334, and 336 relative to SEQ ID NO: 35. In some embodiments, the polypeptide has a sequence identity 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% similar to SEQ ID NO:1295. and a polypeptide comprising an amino acid sequence having sequence identity, except that the polypeptide comprises a set of mutations at positions 60, 61, 63, 64, 69, 74, 76, 85, 112, 115, 116, 119, 120, 121, 130, 134, 145, 146, 154, 190, 198, 223, 225, 227, 231, 241, 251, 258, 273, 274, 278, 289, 303, 314, 334, and 336 compared to SEQ ID NO: 35.In some embodiments, the polypeptide has a sequence similar to that of SEQ ID NO:1301, 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%. % sequence identity to SEQ ID NO: 35, except that the polypeptide comprises a set of mutations at positions 60, 61, 63, 64, 69, 74, 76, 85, 112, 115, 116, 119, 120, 121, 134, 145, 146, 154, 190, 198, 223, 225, 227, 231, 241, 251, 273, 274, 278, 286, 289, 303, 314, 334, and 336 relative to SEQ ID NO: 35. In some embodiments, the polypeptide has a sequence similar to that of SEQ ID NO:1303, 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% sequence identity, except that the polypeptide contains a set of mutations at positions 60, 61, 63, 64, 69, 74, 76, 85, 112, 116, 119, 120, 121, 130, 134, 145, 146, 154, 190, 198, 223, 225, 227, 231, 241, 251, 261, 273, 274, 278, 289, 303, 314, and 334 compared to SEQ ID NO: 35.In some embodiments, the polypeptide has a sequence identity that is 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% similar to SEQ ID NO:1310. and a polypeptide comprising an amino acid sequence having sequence identity, except that the polypeptide comprises a set of mutations at positions 60, 61, 63, 64, 69, 74, 76, 85, 112, 115, 116, 119, 120, 121, 130, 134, 145, 146, 154, 190, 198, 223, 225, 227, 231, 241, 251, 261, 273, 274, 278, 289, 303, 314, 334, and 336 compared to SEQ ID NO: 35.

[0078] In some embodiments, the polypeptide comprises a set of mutations at positions 60, 61, 63, 64, 69, 74, 76, 85, 112, 115, 116, 119, 120, 121, 130, 134, 145, 146, 154, 190, 198, 223, 225, 227, 231, 241, 251, 273, 274, 278, 289, 303, 314, 334, and 336 relative to SEQ ID NO: 35. In some embodiments, the polypeptide comprises a set of mutations at positions 60, 61, 63, 64, 69, 74, 76, 85, 112, 115, 116, 119, 120, 121, 130, 134, 145, 146, 154, 190, 198, 223, 225, 227, 231, 241, 251, 258, 273, 274, 278, 289, 303, 314, 334, 336 compared to SEQ ID NO: 35. In some embodiments, the polypeptide comprises a set of mutations at positions 60, 61, 63, 64, 69, 74, 76, 85, 112, 115, 116, 119, 120, 121, 134, 145, 146, 154, 190, 198, 223, 225, 227, 231, 241, 251, 273, 274, 278, 286, 289, 303, 314, 334, and 336 relative to SEQ ID NO: 35. In some embodiments, the polypeptide comprises a set of mutations at positions 60, 61, 63, 64, 69, 74, 76, 85, 112, 116, 119, 120, 121, 130, 134, 145, 146, 154, 190, 198, 223, 225, 227, 231, 241, 251, 261, 273, 274, 278, 289, 303, 314, and 334 compared to SEQ ID NO: 35. In some embodiments, the polypeptide comprises a set of mutations at positions 60, 61, 63, 64, 69, 74, 76, 85, 112, 115, 116, 119, 120, 121, 130, 134, 145, 146, 154, 190, 198, 223, 225, 227, 231, 241, 251, 261, 273, 274, 278, 289, 303, 314, 334, and 336 relative to SEQ ID NO: 35.

[0079] In some embodiments, the molecule is an Fc 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% sequence identity to SEQ ID NO:1, with the proviso that the Fc polypeptide comprises a mutation or set of mutations at any one position or any combination of positions selected from 234, 235, 237, 296, and 329 relative to SEQ ID NO:1; and an Fc polypeptide having at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80% sequence identity to SEQ ID NO:35. , 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, wherein the polypeptide is 60, 61, 63, 64, 69, 74, 76, 85, 112, 115, 116, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, 135, 136, 137, 138, 139, 140, 141, 142, 143, 144, 145, 146, 147, 148, 149, 150, 151, 152, 153, 154, 155, 156, 157, 158, 159, 200, 2010, 2011, 2012, 2013, 2014, 2015, 2020, 2021, 2022, 2023, 2024, 2025, 2030, 2030, 2031, 2032, 2033, 2034, 2035, 2036, 2037, 2040, 2041, 2042, 2043, 2044, 2045, 2046, 2047, 205 , 130, 134, 145, 146, 154, 190, 198, 223, 225, 227, 231, 241, 251, 258, 261, 273, 274, 278, 286, 289, 303, 314, 334, and 336, and wherein the polypeptide is linked or conjugated to the N-terminus or C-terminus of the Fc polypeptide.

[0080] In some embodiments, the molecule is an Fc 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% sequence identity to SEQ ID NO: 23, with the proviso that the Fc polypeptide comprises a mutation or set of mutations at any one position or any combination of positions selected from 234, 235, 237, 296, and 329 relative to SEQ ID NO: 1; and an Fc polypeptide having at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75% sequence identity to SEQ ID NO: 1290, 1295, 1301, 1303, or 1310. 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity compared to SEQ ID NO: 35, provided that the polypeptide is 60, 61, 63, 64, 69, 74, 76, 85, 112, 115, 116, 119, 200, 201, 202, 203, 204, 205, 206, 207, 208, 209, 210, 211, 212, 213, 214, 215, 216, 217, 218, 219, 220, 221, 222, 223, 224, 225, 226, 227, 228, 229, 230, 231, 232, 233, 234, 235, 236, 237, 238, 239, 240, 241, 242, 243, 244, 245, 246, 247, 248, 249, 250, 251, 252, 253, 254, 255, 256, 257, 258, 259, 260, 261, 262, 263, 264, 265, 266, 267, 268, 269, 300, 301, 302, 303, 304, 305, 306, 307, 308, 309, 310, 311, 3 and a polypeptide comprising a mutation or set of mutations at any one position or any combination of positions selected from 120, 121, 130, 134, 145, 146, 154, 190, 198, 223, 225, 227, 231, 241, 251, 258, 261, 273, 274, 278, 286, 289, 303, 314, 334, and 336, wherein the polypeptide is linked or conjugated to the N-terminus or C-terminus of the Fc polypeptide.

[0081] In some embodiments, the molecule comprises an Fc polypeptide having the amino acid sequence of SEQ ID NO: 23 and a polypeptide having the amino acid sequence of any one of SEQ ID NOs: 1290, 1295, 1301, 1303, or 1310, wherein the polypeptide is linked or conjugated to the N-terminus or C-terminus of the Fc polypeptide.

[0082] In some embodiments, the molecule comprises, from N-terminus to C-terminus, an Fc polypeptide having the amino acid sequence of SEQ ID NO:23 and a polypeptide having the amino acid sequence of SEQ ID NO:1290. In some embodiments, the molecule comprises, from N-terminus to C-terminus, an Fc polypeptide having the amino acid sequence of SEQ ID NO:23 and a polypeptide having the amino acid sequence of SEQ ID NO:1295. In some embodiments, the molecule comprises, from N-terminus to C-terminus, an Fc polypeptide having the amino acid sequence of SEQ ID NO:23 and a polypeptide having the amino acid sequence of SEQ ID NO:1301. In some embodiments, the molecule comprises, N-terminus to C-terminus, an Fc polypeptide having the amino acid sequence of SEQ ID NO:23 and a polypeptide having the amino acid sequence of SEQ ID NO:1303. In some embodiments, the molecule comprises, N-terminus to C-terminus, an Fc polypeptide having the amino acid sequence of SEQ ID NO:23 and a polypeptide having the amino acid sequence of SEQ ID NO:1310. In some embodiments, the molecule comprises, N-terminus to C-terminus, an Fc polypeptide having the amino acid sequence of SEQ ID NO:1290 and an Fc polypeptide having the amino acid sequence of SEQ ID NO:23. In some embodiments, the molecule comprises, from N-terminus to C-terminus, a polypeptide having the amino acid sequence of SEQ ID NO: 1295 and an Fc polypeptide having the amino acid sequence of SEQ ID NO: 23. In some embodiments, the molecule comprises, from N-terminus to C-terminus, a polypeptide having the amino acid sequence of SEQ ID NO: 1301 and an Fc polypeptide having the amino acid sequence of SEQ ID NO: 23. In some embodiments, the molecule comprises, from N-terminus to C-terminus, a polypeptide having the amino acid sequence of SEQ ID NO: 1303 and an Fc polypeptide having the amino acid sequence of SEQ ID NO: 23. In some embodiments, the molecule comprises, N-terminus to C-terminus, a polypeptide having the amino acid sequence of SEQ ID NO: 1310 and an Fc polypeptide having the amino acid sequence of SEQ ID NO: 23.

[0083] In some embodiments, any one mutation or set of mutations selected from A60T, N61D, T63I, Q64K, Q64Y, F69L, V74K, N76G, D112E, K115E, R116N, E119K, E120K, N145D, H146S, E198R, S223N, F227L, E229N, N231T, K241Q, L251I, S273A, N274E, K278E, A303N, K314E, and S334I increases the stability of the polypeptide and reduces or prevents binding of anti-drug antibodies to the polypeptide compared to SEQ ID NO: 35. In some embodiments, a polypeptide comprising any one, or any set of mutations selected from A60T, N61D, T63I, Q64K, Q64Y, F69L, V74K, N76G, D112E, K115E, R116N, E119K, E120K, N145D, H146S, E198R, S223N, F227L, E229N, N231T, K241Q, L251I, S273A, N274E, K278E, A303N, K314E, and S334I has increased stability and / or reduced or no binding to anti-drug antibodies compared to SEQ ID NO: 35.

[0084] In some embodiments, the polypeptide comprises any one of the following amino acids: A60T, N61D, T63I, Q64Y, F69L, V74K, N76G, T85L, D112E, K115E, R116N, E119K, E120K, H121Y, N130K, N130G, M134L, N145D, H146S, E154D, N190K, H191Y, N131K, N131G, M134L, N145D, H146S, E154D, N190K, H191Y, N132K, N132G, M134L, N145D, H146S, E154D, N190K, H191Y, N133K, N133G, M134L, N1 The mutation or set of mutations may be selected from any one or any combination of: D, E198R, S223N, H225Y, F227L, N231T, K241Q, L251I, V258G, N261G, S273A, N274E, K278E, D286N, A289P, A303S, K314E, S334I, and N336Q. In some embodiments, the polypeptide comprises the following set of mutations compared to SEQ ID NO: 35: A60T, N61D, T63I, Q64Y, F69L, V74K, N76G, T85L, D112E, K115E, R116N, E119K, E120K, H121Y, N130K, M134L, N145D, H146S, E154D, N190D, E198R, S223N, H225Y, F227L, N231T, K241Q, L251I, S273A, N274E, K278E, A289P, A303S, K314E, S334I, and N336Q. In some embodiments, the polypeptide comprises the following set of mutations compared to SEQ ID NO: 35: A60T, N61D, T63I, Q64Y, F69L, V74K, N76G, T85L, D112E, K115E, R116N, E119K, E120K, H121Y, N130K, M134L, N145D, H146S, E154D, N190D, E198R, S223N, H225Y, F227L, N231T, K241Q, L251I, V258G, S273A, N274E, K278E, A289P, A303S, K314E, S334I, and N336Q.In some embodiments, the polypeptide comprises the following set of mutations compared to SEQ ID NO: 35: A60T, N61D, T63I, Q64Y, F69L, V74K, N76G, T85L, D112E, K115E, R116N, E119K, E120K, H121Y, M134L, N145D, H146S, E154D, N190D, E198R, S223N, H225Y, F227L, N231T, K241Q, L251I, S273A, N274E, K278E, D286N, A289P, A303S, K314E, S334I, and N336Q. In some embodiments, the polypeptide comprises the following set of mutations compared to SEQ ID NO: 35: A60T, N61D, T63I, Q64Y, F69L, V74K, N76G, T85L, D112E, R116N, E119K, E120K, H121Y, N130G, M134L, N145D, H146S, E154D, N190D, E198R, S223N, H225Y, F227L, N231T, K241Q, L251I, N261G, S273A, N274E, K278E, A289P, A303S, K314E, and S334I. In some embodiments, the polypeptide comprises the following set of mutations compared to SEQ ID NO: 35: A60T, N61D, T63I, Q64Y, F69L, V74K, N76G, T85L, D112E, K115E, R116N, E119K, E120K, H121Y, N130K, M134L, N145D, H146S, E154D, N190D, E198R, S223N, H225Y, F227L, N231T, K241Q, L251I, N261G, S273A, N274E, K278E, A289P, A303S, K314E, S334I, and N336Q.

[0085] In some embodiments, the polypeptide comprises 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% sequence identity to SEQ ID NO: 1290, with the proviso that the polypeptide comprises the following set of mutations compared to SEQ ID NO: 35: A60T, N61D, T63I, Q64Y, F69L, V74K, N76G, T85L, D112E, K115E, R116N, E119K, E120K, H121Y, N130K, M134L, N145D, H146S, E154D, N190D, E198R, S223N, H225Y, F227L, N231T, K241Q, L251I, S273A, N274E, K278E, A289P, A303S, K314E, S334I, and N336Q. In some embodiments, the polypeptide comprises 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% sequence identity to SEQ ID NO: 1295, provided that the polypeptide does not comprise a sequence Compared to column number 35, the mutation set includes A60T, N61D, T63I, Q64Y, F69L, V74K, N76G, T85L, D112E, K115E, R116N, E119K, E120K, H121Y, N130K, M134L, N145D, H146S, E154D, N190D, E198R, S223N, H225Y, F227L, N231T, K241Q, L251I, V258G, S273A, N274E, K278E, A289P, A303S, K314E, S334I, and N336Q.In some embodiments, the polypeptide comprises 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% sequence identity to SEQ ID NO: 1301, with the proviso that the polypeptide comprises the following set of mutations compared to SEQ ID NO: 35: A60T, N61D, T63I, Q64Y, F69L, V74K, N76G, T85L, D112E, K115E, R116N, E119K, E120K, H121Y, M134L, N145D, H146S, E154D, N190D, E198R, S223N, H225Y, F227L, N231T, K241Q, L251I, S273A, N274E, K278E, D286N, A289P, A303S, K314E, S334I, and N336Q. In some embodiments, the polypeptide comprises 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% sequence identity to SEQ ID NO: 1303, provided that the polypeptide The peptide comprises the following set of mutations compared to SEQ ID NO: 35: A60T, N61D, T63I, Q64Y, F69L, V74K, N76G, T85L, D112E, R116N, E119K, E120K, H121Y, N130G, M134L, N145D, H146S, E154D, N190D, E198R, S223N, H225Y, F227L, N231T, K241Q, L251I, N261G, S273A, N274E, K278E, A289P, A303S, K314E, and S334I.In some embodiments, the polypeptide comprises 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% sequence identity to SEQ ID NO: 1310, provided that the polypeptide does not comprise a sequence Compared to column number 35, the mutation set includes A60T, N61D, T63I, Q64Y, F69L, V74K, N76G, T85L, D112E, K115E, R116N, E119K, E120K, H121Y, N130K, M134L, N145D, H146S, E154D, N190D, E198R, S223N, H225Y, F227L, N231T, K241Q, L251I, N261G, S273A, N274E, K278E, A289P, A303S, K314E, S334I, and N336Q.

[0086] In some embodiments, the polypeptide has a sequence identity 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%, or at least 50% of any one of SEQ ID NOs: 1303, 1290, 1291, 1292, 1293, 1294, 1295, 1296, 1297, 1298, 1299, 1300, 1301, 1302, 1304, 1305, 1306, 1307, 1308, 1309, 1310, and 1333. and a polypeptide having an amino acid sequence having at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity, relative to SEQ ID NO:35, wherein the polypeptide contains at least one of the following amino acids: threonine (T) at position 60, aspartic acid (D) at position 61, isoleucine (I) at position 63, tyrosine (Y) at position 64, leucine (L) at position 69, lysine (K) at position 74, glycine (G) at position 76, leucine (L) at position 85, glutamic acid (E) at position 112, and a nucleotide sequence identical to SEQ ID NO:35. Glutamic acid (E) at position 115, asparagine (N) at position 116, lysine (K) at position 119, lysine (K) at position 120, tyrosine (Y) at position 121, lysine (K) at position 130, glycine (G) at position 130, leucine (L) at position 134, aspartic acid (D) at position 145, serine (S) at position 146, aspartic acid (D) at position 154, aspartic acid (D) at position 190, arginine (R) at position 198, asparagine (N) at position 223, tyrosine (Y) at position 225, The amino acid sequence may include any one or any combination of leucine (L), threonine (T) at position 231, glutamine (Q) at position 241, isoleucine (I) at position 251, glycine (G) at position 261, alanine (A) at position 273, glutamic acid (E) at position 274, glutamic acid (E) at position 278, asparagine (N) at position 286, proline (P) at position 289, serine (S) at position 303, glutamic acid (E) at position 314, isoleucine (I) at position 334, and glutamine (Q) at position 336.

[0087] In some embodiments, the polypeptide comprises 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 SEQ ID NO: 1303, except that the polypeptide contains the following amino acid residues relative to 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 threonine (T) at position 67, an aspartic acid (D) at position 68, an isoleucine (I) at position 69, a tyrosine (Y) at position 71, a lysine (K) at position 72, a glycine (G) at position 73, a leucine (L) at position 85, a glutamic acid (E) at position 112, an asparagine (N) at position 116, a threonine (T) at position 118, a threonine (T) at position 119, a threonine (T) at position 120, a threonine (T) at position 122, a threonine (T) at position 124, a threonine (T) at position 126, a threonine lysine (K) at position 9, lysine (K) at position 120, tyrosine (Y) at position 121, glycine (G) at position 130, leucine (L) at position 134, aspartic acid (D) at position 145, serine (S) at position 146, aspartic acid (D) at position 154, aspartic acid (D) at position 190, arginine (R) at position 198, asparagine (N) at position 223, tyrosine (Y) at position 225, and leucine (L) at position 227. It contains leucine (L), threonine (T) at position 231, glutamine (Q) at position 241, isoleucine (I) at position 251, glycine (G) at position 261, alanine (A) at position 273, glutamic acid (E) at position 274, glutamic acid (E) at position 278, cysteine ​​(P) at position 289, serine (S) at position 303, glutamic acid (E) at position 314, and isoleucine (I) at position 334.

[0088] As used herein, the terms "corresponding to a position," "compared to" a reference sequence, or similar terms, mean that the residues line up when the two sequences are aligned. For example, Figure 9 shows an alignment of a non-limiting polypeptide provided herein (e.g., SEQ ID NO: 1303) with SEQ ID NO: 35, showing how the positions correspond to / align with the positions in SEQ ID NO: 35. The alignment was performed using clustal omega and default settings. Clustal omega can be accessed, for example, at www"dot"ebi"dot"ac"dot"uk / Tools / msa / clustalo / . Thus, the mutations A60T, N61D, T63I, Q64Y, F69L, V74K, N76G, T85L, D112E, K115E, R116N, E119K, E120K, H121Y, N130K, N130G, M134L, N145D, H146S, E154D, N190D, E198R, S223N, H225Y, F227L, N231T, K241Q, L251I, V258G, N261G, S273A, N274E, K278E, D286N, A289P, A303S, K314E, S334I, and N336Q are compared to the wild-type full-length IdeS amino acid sequence of 35. In some embodiments, the mutations are relative to the polypeptides disclosed herein, which may be variants and / or truncations of wild-type full-length IdeS having the amino acid sequence of SEQ ID NO:35.In some embodiments, the amino acid sequence of any of the amino acids selected from the group consisting of A60T, N61D, T63I, Q64Y, F69L, V74K, N76G, T85L, D112E, K115E, R116N, E119K, E120K, H121Y, N130K, N130G, M134L, N145D, H146S, E154D, N190D, E198R, S223N, H225Y, F227L, N231T, K241Q, L251I, V258G, N261G, S273A, N274E, K278E, D286N, A289P, A303S, K314E, S334I, and N336Q as compared to SEQ ID NO: 35. The mutations may be provided as, for example, A13T, N14T, T16I, Q17Y, F22L, V27K, N29G, T38L, D65E, K68E, R69N, E72K, E73K, H74Y, N83K, N83G, M87L, N98D, H99S, E107D, N143D, E151R, S176N, H178Y, F180L, N184T, K194Q, L204I, V211G, N214G, S226A, N227E, K231E, D239N, A242P, A256S, K267E, S287I, and N289Q, respectively, compared to SEQ ID NO: 1303. Thus, in some embodiments, the polypeptide comprises 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 SEQ ID NO: 1303, provided that the polypeptide has at least one amino acid sequence that is ... The set of mutations includes 3T, N14T, T16I, Q17Y, F22L, V27K, N29G, T38L, D65E, K68E, R69N, E72K, E73K, H74Y, N83K, N83G, M87L, N98D, H99S, E107D, N143D, E151R, S176N, H178Y, F180L, N184T, K194Q, L204I, V211G, N214G, S226A, N227E, K231E, D239N, A242P, A256S, K267E, S287I, and N289Q.

[0089] In some embodiments, the polypeptide comprises 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 SEQ ID NO: 1303, provided that the polypeptide contains at least one of the following amino acids: a threonine (T) at position 13, an aspartic acid (D) at position 14, an isoleucine (I) at position 16, a tyrosine (Y) at position 17, a leucine (L) at position 22, a lysine (K) at position 27, a glycine (G) at position 29, a leucine (L) at position 38, a glutamic acid (E) at position 65, an asparagine (N) at position 69, ... Lysine (K) at position 72, Lysine (K) at position 73, Tyrosine (Y) at position 74, Glycine (G) at position 83, Leucine (L) at position 87, Aspartic acid (D) at position 98, Serine (S) at position 99, Aspartic acid (D) at position 107, Aspartic acid (D) at position 143, Arginine (R) at position 151, Asparagine (N) at position 176, Tyrosine (Y) at position 178, Leucine (L) at position 180 It contains a leucine (L) at position 184, a threonine (T) at position 184, a glutamine (Q) at position 194, an isoleucine (I) at position 204, a glycine (G) at position 214, an alanine (A) at position 226, a glutamic acid (E) at position 227, a glutamic acid (E) at position 231, a cysteine ​​(P) at position 242, a serine (S) at position 256, a glutamic acid (E) at position 267, and an isoleucine (I) at position 287.

[0090] In some embodiments, the polypeptide has a sequence identity that is 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% of any one of SEQ ID NOs: 1290, 1291, 1292, 1293, 1294, 1295, 1296, 1297, 1298, 1299, 1300, 1301, 1302, 1304, 1305, 1306, 1307, 1308, 1309, 1310, and 1333. , or an amino acid sequence having at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO:35, except that the polypeptide contains at least one of the following amino acids: 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, a glutamic acid (E) at position 115, an asparagine (N) at position 116, a lysine (K) at position 119, a threonine (T) at position 61 ... Lysine (K) at position 20, tyrosine (Y) at position 121, lysine (K) at position 130, leucine (L) at position 134, aspartic acid (D) at position 145, serine (S) at position 146, aspartic acid (D) at position 154, aspartic acid (D) at position 190, arginine (R) at position 198, asparagine (N) at position 223, tyrosine (Y) at position 225, leucine (L) at position 227, threonine (T) at position 231, glutamine (Q) at position 241, isoleucine (I) at position 251, alanine (A) at position 273, glutamic acid ( E), glutamic acid (E) at position 278, proline (P) at position 289, serine (S) at position 303, glutamic acid (E) at position 314, isoleucine (I) at position 334, and glutamine (Q) at position 336; compared to SEQ ID NO: 35, threonine (T) at position 60, aspartic acid (D) at position 61, isoleucine (I) at position 63, tyrosine (Y) at position 64, leucine (L) at position 69, lysine (K) at position 74, glycine (G) at position 76, leucine (L) at position 85, glutamic acid (E) at position 112, glutamic acid (E) at position 115;Asparagine (N) at position 116, lysine (K) at position 119, lysine (K) at position 120, tyrosine (Y) at position 121, lysine (K) at position 130, leucine (L) at position 134, aspartic acid (D) at position 145, serine (S) at position 146, aspartic acid (D) at position 154, aspartic acid (D) at position 190, arginine (R) at position 198, asparagine (N) at position 223, tyrosine (Y) at position 225, leucine (L) at position 227, threonine (T) at position 231, glutamine (Q) at position 241, and isoleucine (I) at position 251. ), glycine (G) at position 258, alanine (A) at position 273, glutamic acid (E) at position 274, glutamic acid (E) at position 278, proline (P) at position 289, serine (S) at position 303, glutamic acid (E) at position 314, isoleucine (I) at position 334, and glutamine (Q) at position 336; compared to SEQ ID NO: 35, threonine (T) at position 60, aspartic acid (D) at position 61, isoleucine (I) at position 63, tyrosine (Y) at position 64, leucine (L) at position 69, lysine (K) at position 74, glycine (G) at position 85 Leucine (L) at position 112, glutamic acid (E) at position 115, asparagine (N) at position 116, lysine (K) at position 119, lysine (K) at position 120, tyrosine (Y) at position 121, leucine (L) at position 134, aspartic acid (D) at position 145, serine (S) at position 146, aspartic acid (D) at position 154, aspartic acid (D) at position 190, arginine (R) at position 198, asparagine (N) at position 223, tyrosine (Y) at position 225, leucine (L) at position 227, threonine (T) at position 231, glutamine (Q) at position 241, isoleucine (I) at position 251, alanine (A) at position 273, glutamic acid (E) at position 274, glutamic acid (E) at position 278, asparagine (N) at position 286, proline (P) at position 289, serine (S) at position 303, glutamic acid (E) at position 314, isoleucine (I) at position 334, and glutamine (Q) at position 336; compared to SEQ ID NO: 35, threonine (T) at position 60, aspartic acid (D) at position 61, isoleucine (I) at position 63, tyrosine (Y) at position 64, leucine (L) at position 69;Lysine (K) at position 74, glycine (G) at position 76, leucine (L) at position 85, glutamic acid (E) at position 112, asparagine (N) at position 116, lysine (K) at position 119, lysine (K) at position 120, tyrosine (Y) at position 121, glycine (G) at position 130, leucine (L) at position 134, aspartic acid (D) at position 145, serine (S) at position 146, aspartic acid (D) at position 154, aspartic acid (D) at position 190, arginine (R) at position 198, asparagine (N) at position 223, tyrosine (Y) at position 225, Leucine (L) at position 227, threonine (T) at position 231, glutamine (Q) at position 241, isoleucine (I) at position 251, glycine (G) at position 261, alanine (A) at position 273, glutamic acid (E) at position 274, glutamic acid (E) at position 278, proline (P) at position 289, serine (S) at position 303, glutamic acid (E) at position 314, and isoleucine (I) at position 334, or, compared to SEQ ID NO: 35, threonine (T) at position 60, aspartic acid (D) at position 61, isoleucine (I) at position 63, tyrosine (G) at position 64, and glycine (G) at position 261. lysine (K) at position 112, glycine (G) at position 116, leucine (L) at position 69, lysine (K) at position 74, glycine (G) at position 76, leucine (L) at position 85, glutamic acid (E) at position 112, glutamic acid (E) at position 115, asparagine (N) at position 116, lysine (K) at position 119, lysine (K) at position 120, tyrosine (Y) at position 121, lysine (K) at position 130, leucine (L) at position 134, aspartic acid (D) at position 145, serine (S) at position 146, aspartic acid (D) at position 154, aspartic acid (D) at position 190, arginine (R) at position 198 ), asparagine (N) at position 223, tyrosine (Y) at position 225, leucine (L) at position 227, threonine (T) at position 231, glutamine (Q) at position 241, isoleucine (I) at position 251, glycine (G) at position 261, alanine (A) at position 273, glutamic acid (E) at position 274, glutamic acid (E) at position 278, proline (P) at position 289, serine (S) at position 303, glutamic acid (E) at position 314, isoleucine (I) at position 334, and glutamine (Q) at position 336.

[0091] In some embodiments, the polypeptide contains the following amino acids, relative to SEQ ID NO:35: threonine (T) at position 60, aspartic acid (D) at position 61, isoleucine (I) at position 63, tyrosine (Y) at position 64, leucine (L) at position 69, lysine (K) at position 74, glycine (G) at position 76, leucine (L) at position 85, glutamic acid (E) at position 112, glutamic acid (E) at position 115, asparagine (N) at position 116, lysine (K) at position 119, lysine (K) at position 120, tyrosine (Y) at position 121, lysine (K) at position 130, lysine (K) at position 13 ...136, leucine (L) at position 138, leucine (L) at position 139, leucine (L) at position 140, leucine (L) at position 142, leucine (L) at position 143, leucine (L) at position 144, leucine (L) at position 14 isoleucine (L), aspartic acid (D) at position 145, serine (S) at position 146, aspartic acid (D) at position 154, aspartic acid (D) at position 190, arginine (R) at position 198, asparagine (N) at position 223, tyrosine (Y) at position 225, leucine (L) at position 227, threonine (T) at position 231, glutamine (Q) at position 241, isoleucine (I) at position 251, alanine (A) at position 273, glutamic acid (E) at position 274, glutamic acid (E) at position 278, proline (P) at position 289, serine (S) at position 303, glutamic acid (E) at position 314, isoleucine (I) at position 334, and glutamine (Q) at position 336; compared to SEQ ID NO: 35, threonine (T) at position 60, aspartic acid (D) at position 61, isoleucine (I) at position 63, tyrosine (Y) at position 64, leucine (L) at position 69, lysine (K) at position 74, glycine (G) at position 76, leucine (L) at position 85, glutamic acid (E) at position 112, glutamic acid (E) at position 115, asparagine (N) at position 116, lysine (K) at position 119, lysine (K) at position 120, glycine (G) at position 121, leucine (L) at position 122, glutamic acid (E) at position 115, asparagine (N) at position 116, lysine (K) at position 119, lysine (K) at position 121, glycine (G) at position 122, leucine (L) at position 123, leucine (L) at position 124, leucine (L) at position 125, glutamic acid (E) at position 115, glutamic acid (E) at position 116, asparagine (N) at position 116, lysine (K) at position 119, lysine (K) at position 120, glycine (G) at position 121, leucine (L) at position 122, leucine (L) at position 123, leucine (L) at position 124, leucine (L) at position 125, leucine (L) at tyrosine (Y) at position 130, lysine (K) at position 134, leucine (L) at position 145, aspartic acid (D) at position 146, serine (S) at position 146, aspartic acid (D) at position 154, aspartic acid (D) at position 190, arginine (R) at position 198, asparagine (N) at position 223, tyrosine (Y) at position 225, leucine (L) at position 227, threonine (T) at position 231, glutamine (Q) at position 241, isoleucine (I) at position 251, glycine (G) at position 258, alanine (A) at position 273, glutamic acid (E) at position 274,glutamic acid (E) at position 278, proline (P) at position 289, serine (S) at position 303, glutamic acid (E) at position 314, isoleucine (I) at position 334, and glutamine (Q) at position 336; compared to SEQ ID NO: 35, threonine (T) at position 60, aspartic acid (D) at position 61, isoleucine (I) at position 63, tyrosine (Y) at position 64, leucine (L) at position 69, lysine (K) at position 74, glycine (G) at position 76, leucine (L) at position 85, glutamic acid (E) at position 112, glutamic acid (E) at position 115, Asparagine (N) at position 6, Lysine (K) at position 119, Lysine (K) at position 120, Tyrosine (Y) at position 121, Leucine (L) at position 134, Aspartic acid (D) at position 145, Serine (S) at position 146, Aspartic acid (D) at position 154, Aspartic acid (D) at position 190, Arginine (R) at position 198, Asparagine (N) at position 223, Tyrosine (Y) at position 225, Leucine (L) at position 227, Threonine (T) at position 231, Glutamine (Q) at position 241, Isoleucine (I) at position 251, Alanine (A) at position 273, glutamic acid (E) at position 274, glutamic acid (E) at position 278, asparagine (N) at position 286, proline (P) at position 289, serine (S) at position 303, glutamic acid (E) at position 314, isoleucine (I) at position 334, and glutamine (Q) at position 336; compared to SEQ ID NO: 35, threonine (T) at position 60, aspartic acid (D) at position 61, isoleucine (I) at position 63, tyrosine (Y) at position 64, leucine (L) at position 69, lysine (K) at position 74, glycine (G) at position 76, leucine (L) at position 85, leucine (L) at position 11 Glutamic acid (E) at position 2, asparagine (N) at position 116, lysine (K) at position 119, lysine (K) at position 120, tyrosine (Y) at position 121, glycine (G) at position 130, leucine (L) at position 134, aspartic acid (D) at position 145, serine (S) at position 146, aspartic acid (D) at position 154, aspartic acid (D) at position 190, arginine (R) at position 198, asparagine (N) at position 223, tyrosine (Y) at position 225, leucine (L) at position 227, threonine (T) at position 231, glutamine (Q) 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 serine (S) at position 303, a glutamic acid (E) at position 314, and an isoleucine (I) at position 334; or, compared to SEQ ID NO: 35, a threonine (T) at position 60, an asparagine (A) at position 61, an alanine (A) at position 273, an alanine (A) at position 274, an alanine (A) at position 278, a proline (P) at position 289, a serine (S) at position 303, an asparagine (A) at position 314, and an isoleucine (I) at position 334; glycine (D), isoleucine (I) at position 63, tyrosine (Y) at position 64, leucine (L) at position 69, lysine (K) at position 74, glycine (G) at position 76, leucine (L) at position 85, glutamic acid (E) at position 112, glutamic acid (E) at position 115, asparagine (N) at position 116, lysine (K) at position 119, lysine (K) at position 120, tyrosine (Y) at position 121, Lysine (K) at position 130, leucine (L) at position 134, aspartic acid (D) at position 145, serine (S) at position 146, aspartic acid (D) at position 154, aspartic acid (D) at position 190, arginine (R) at position 198, asparagine (N) at position 223, tyrosine (Y) at position 225, leucine (L) at position 227, threonine (T) at position 231, glutamine ( and a proline (P) at position 289; a serine (S) at position 303; a glutamic acid (E) at position 314; an isoleucine (I) at position 334; and a glutamine (Q) at position 336.

[0092] In some embodiments, a polypeptide is provided that is at least 50%, 60%, 70%, 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to a polypeptide comprising the amino acid sequence of SEQ ID NO: 36 or 1349, provided that the polypeptide is Threonine (T) at position 60, aspartic acid (D) at position 61, isoleucine (I) at position 63, tyrosine (Y) at position 64, leucine (L) at position 69, lysine (K) at position 74, glycine (G) at position 76, leucine (L) at position 85, glutamic acid (E) at position 112, asparagine (N) at position 116, lysine (K) at position 119, lysine (K) at position 120, tyrosine ( Glycine (G) at position 130, leucine (L) at position 134, aspartic acid (D) at position 145, serine (S) at position 146, aspartic acid (D) at position 154, aspartic acid (D) at position 190, arginine (R) at position 198, asparagine (N) at position 223, tyrosine (Y) at position 225, leucine (L) at position 227, threonine (T) at position 231, glutamic acid (G) at position 241, and one or more or all of: alanine (A) at position 273, glutamic acid (E) at position 274, glutamic acid (E) at position 278, proline (P) at position 289, serine (S) at position 303, glutamic acid (E) at position 314, and isoleucine (I) at position 334, which positions correspond to those in SEQ ID NO: 35. In some embodiments, a polypeptide is at least 80% identical to a polypeptide comprising the amino acid sequence of SEQ ID NO: 36 or 1349, under the proviso set forth in this paragraph. In some embodiments, a polypeptide is at least 81% identical to a polypeptide comprising the amino acid sequence of SEQ ID NO: 36 or 1349, under the proviso set forth in this paragraph. In some embodiments, a polypeptide is at least 82% identical to a polypeptide comprising the amino acid sequence of SEQ ID NO: 36 or 1349, under the proviso set forth in this paragraph.In some embodiments, the polypeptide is at least 83% identical to a polypeptide comprising the amino acid sequence of SEQ ID NO: 36 or 1349, according to the proviso set forth in this paragraph. In some embodiments, the polypeptide is at least 84% identical to a polypeptide comprising the amino acid sequence of SEQ ID NO: 36 or 1349, according to the proviso set forth in this paragraph. In some embodiments, the polypeptide is at least 85% identical to a polypeptide comprising the amino acid sequence of SEQ ID NO: 36 or 1349, according to the proviso set forth in this paragraph. In some embodiments, the polypeptide is at least 86% identical to a polypeptide comprising the amino acid sequence of SEQ ID NO: 36 or 1349, according to the proviso set forth in this paragraph. In some embodiments, the polypeptide is at least 87% identical to a polypeptide comprising the amino acid sequence of SEQ ID NO: 36 or 1349, according to the proviso set forth in this paragraph. In some embodiments, the polypeptide is at least 88% identical to a polypeptide comprising the amino acid sequence of SEQ ID NO: 36 or 1349, according to the proviso set forth in this paragraph. In some embodiments, the polypeptide is at least 89% identical to a polypeptide comprising the amino acid sequence of SEQ ID NO: 36 or 1349, according to the proviso set forth in this paragraph. In some embodiments, the polypeptide is at least 90% identical to a polypeptide comprising the amino acid sequence of SEQ ID NO: 36 or 1349, subject to the proviso described in this paragraph. In some embodiments, the polypeptide does not comprise the amino acid sequence of DSFSANQEIRYSEVTPYHVT (SEQ ID NO: 1351). In some embodiments, the polypeptide does not comprise the amino acid sequence of DSFSANQEIRYSEVTPYHVT (SEQ ID NO: 1351). In some embodiments, the polypeptide does not comprise the entire consecutive amino acid sequence of DSFSANQEIRYSEVTPYHVT (SEQ ID NO: 1351). In some embodiments, the polypeptide does not comprise the entire consecutive amino acid sequence of DSFSANQEIRYSEVTPYHVT (SEQ ID NO: 1351) or does not comprise the VT dipeptide at the N-terminus of the polypeptide.As used herein, the N-terminus of a polypeptide with respect to the consecutive amino acid sequence of DSFSANQEIRYSEVTPYHVT (SEQ ID NO: 1351), or the dipeptide of VT, refers to the N-terminus of the protease, which may be the C-terminus of a linker or another polypeptide when the protease is linked to another polypeptide, such as an Fc domain provided herein. In some embodiments, a polypeptide does not comprise the dipeptide of VT at the N-terminus of the polypeptide. In some embodiments, a polypeptide does not comprise the amino acid sequence of DSFSANQEIRYSEVTPYH (SEQ ID NO: 1352). In some embodiments, a polypeptide does not comprise the amino acid sequence of ANQEIRYSEVTPYHVT (SEQ ID NO: 1353). In some embodiments, a polypeptide does not comprise the amino acid sequence of ANQEIRYSEVTPYH (SEQ ID NO: 1354). In some embodiments, a polypeptide does not comprise the amino acid sequence of NQTN (SEQ ID NO: 1355). In some embodiments, a polypeptide does not comprise the amino acid sequence of NQTN (SEQ ID NO: 1355). In some embodiments, a polypeptide does not comprise the consecutive amino acid sequence of NQTN (SEQ ID NO: 1355) at the C-terminus of the polypeptide. As used herein, the C-terminus of a polypeptide with respect to the contiguous amino acid sequence of NQTN (SEQ ID NO: 1355) refers to the C-terminus of the protease, which may be the N-terminus of a linker or another polypeptide when the protease is linked to another polypeptide, such as an Fc domain provided herein. In some embodiments, the polypeptide comprises the sequence of QQTN (SEQ ID NO: 1356) at the C-terminus of the polypeptide instead of the sequence of NQTN (SEQ ID NO: 1355).

[0093] In some embodiments, the polypeptide has a cysteine ​​(C) at a position in the polypeptide corresponding to position 94 of SEQ ID NO:35. In some embodiments, the polypeptide has a lysine (K), a histidine (H), an aspartic acid (D), and an aspartic acid (D) at positions in the variant sequence corresponding to positions 84, 262, 284, and 286 of SEQ ID NO:35, 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 less immunogenic than an IdeS polypeptide, such as an IdeS polypeptide comprising the amino acid sequence of SEQ ID NO:35, SEQ ID NO:36, or SEQ ID NO:1349. In some embodiments, the polypeptide produces greater amounts of IgG cleavage fragments than IdeS (e.g., SEQ ID NO:35, SEQ ID NO:36, or SEQ ID NO:1349). In some embodiments, the polypeptide has a positively charged amino acid, such as arginine (R) or lysine (K), at a position in the variant corresponding to position 130 of SEQ ID NO:35. In some embodiments, the polypeptide has a positively charged amino acid, such as arginine (R) or lysine (K), at a position in the variant corresponding to position 131 of SEQ ID NO:35.

[0094] 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 C-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 via a linker. In some embodiments, the protease-resistant Fc domain is linked to the polypeptide via 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, lysine (K), arginine (R), or histidine (H). In some embodiments, the peptide linker comprises the amino acid sequence of SEQ ID NO: 1330. In some embodiments, the protease-resistant Fc domain is as provided herein. In some embodiments, the protease resistant Fc domain comprises the sequence of any one of SEQ ID NOs: 19, 20, 21, 22, 23, 24, 25, 42, 43, 44, 45, 46, 47, 48, or 49. In some embodiments, the protease resistant Fc domain comprises the amino acid sequence of SEQ ID NO: 23. In some embodiments, the polypeptide comprises, from N-terminus to C-terminus, a polypeptide, 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.

[0095] In some embodiments, the polypeptide comprises the amino acid sequence shown in Table 7 in the column labeled "Full Length Sequence" for VRT-1 through VRT-319. 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, which optionally comprises the amino acid sequence of SEQ ID NO: 35, SEQ ID NO: 36, or SEQ ID NO: 1349. In some embodiments, the polypeptide produces greater amounts of IgG cleavage fragments than IdeS (e.g., SEQ ID NO: 35, SEQ ID NO: 36, or SEQ ID NO: 1349). In some embodiments, the polypeptide of SEQ ID NO: 1349 produces greater amounts of IgG cleavage fragments than IdeS.

[0096] In some embodiments, the H225Y mutation increases expression of the polypeptide compared to SEQ ID NO: 35. In some embodiments, a polypeptide comprising the H225Y mutation compared to SEQ ID NO: 35 has increased expression. In some embodiments, any one or set of mutations selected from T85L, H121Y, E154D, N190D, A289P, and N303S compared to SEQ ID NO: 35 eliminates a T cell binding epitope. In some embodiments, any one or set of mutations selected from T85L, H121Y, E154D, N190D, A289P, and N303S compared to SEQ ID NO: 35 reduces or prevents T cell binding to the polypeptide. In some embodiments, a polypeptide comprising any one or any set of mutations selected from T85L, H121Y, E154D, N190D, A289P, and N303S compared to SEQ ID NO: 35 does not have a T cell binding epitope. In some embodiments, a polypeptide comprising any one or any set of mutations selected from T85L, H121Y, E154D, N190D, A289P, and N303S relative to SEQ ID NO: 35 does not bind to T cells. In some embodiments, any one or set of mutations selected from T85L, H121Y, E154D, N190D, A289P, and N303S relative to SEQ ID NO: 35 eliminates a T cell binding epitope. In some embodiments, any one or set of mutations selected from T85L, H121Y, E154D, N190D, A289P, and N303S relative to SEQ ID NO: 35 reduces or prevents T cell binding to the polypeptide. In some embodiments, a polypeptide comprising any one, or any set of mutations selected from T85L, H121Y, E154D, N190D, V258G, A289P, and N303S relative to SEQ ID NO: 35 does not have a T cell binding epitope. In some embodiments, a polypeptide comprising any one, or any set of mutations selected from T85L, H121Y, E154D, N190D, V258G, A289P, and N303S relative to SEQ ID NO: 35 does not bind to T cells.

[0097] In some embodiments, any one mutation or set of mutations selected from M134L, N190D, N229E, A289P, and N336Q relative to SEQ ID NO: 35 eliminates chemical liability. In some embodiments, a polypeptide comprising any one or any set of mutations selected from M134L, N190D, N229E, A289P, and N336Q relative to SEQ ID NO: 35 is non-chemically liable. In some embodiments, any one mutation or set of mutations selected from N130K, M134L, N190D, N229E, A289P, and N336Q relative to SEQ ID NO: 35 eliminates chemical liability. In some embodiments, a polypeptide comprising any one or any set of mutations selected from N130K, M134L, N190D, N229E, A289P, and N336Q relative to SEQ ID NO: 35 is non-chemically liable.

[0098] In some embodiments, any one or set of mutations selected from T85L, H121Y, N130K, M134L, E154D, N190D, N229E, A289P, N303S, and N336Q relative to SEQ ID NO: 35 increases the stability of the polypeptide, reduces or prevents binding of anti-drug antibodies to the polypeptide, eliminates a T-cell binding epitope, and eliminates chemical liability. In some embodiments, any one or set of mutations selected from T85L, H121Y, N130K, M134L, E154D, N190D, N229E, A289P, N303S, and N336Q relative to SEQ ID NO: 35 increases the stability of the polypeptide, reduces or prevents binding of anti-drug antibodies to the polypeptide, reduces or prevents T-cell binding to the polypeptide, and / or eliminates chemical liability. In some embodiments, a polypeptide comprising any one or any set of mutations selected from T85L, H121Y, N130K, M134L, E154D, N190D, N229E, A289P, N303S, and N336Q relative to SEQ ID NO: 35 has increased stability, reduced or no binding to anti-drug antibodies, no T-cell binding epitopes, and / or no chemosensitivity. In some embodiments, a polypeptide comprising any one or any set of mutations selected from T85L, H121Y, N130K, M134L, E154D, N190D, N229E, A289P, N303S, and N336Q relative to SEQ ID NO: 35 has increased stability, reduced or no binding to anti-drug antibodies, no T-cell binding epitopes, and / or no chemosensitivity.

[0099] In some embodiments, any one or set of mutations selected from T85L, H121Y, M134L, E154D, N190D, N229E, A289P, N303S, and N336Q relative to SEQ ID NO: 35 increases the stability of the polypeptide, reduces or prevents binding of anti-drug antibodies to the polypeptide, eliminates a T-cell binding epitope, and eliminates chemical liability. In some embodiments, any one or set of mutations selected from T85L, H121Y, M134L, E154D, N190D, N229E, A289P, N303S, and N336Q relative to SEQ ID NO: 35 increases the stability of the polypeptide, reduces or prevents binding of anti-drug antibodies to the polypeptide, reduces or prevents T-cell binding to the polypeptide, and / or eliminates chemical liability. In some embodiments, a polypeptide comprising any one or any set of mutations selected from T85L, H121Y, M134L, E154D, N190D, N229E, A289P, N303S, and N336Q relative to SEQ ID NO: 35 has increased stability, reduced or no binding to anti-drug antibodies, no T-cell binding epitopes, and / or no chemosensitivity. In some embodiments, a polypeptide comprising any one or any set of mutations selected from T85L, H121Y, M134L, E154D, N190D, N229E, A289P, N303S, and N336Q relative to SEQ ID NO: 35 has increased stability, reduced or no binding to anti-drug antibodies, no T-cell binding epitopes, and / or no chemosensitivity. In some embodiments, any one or set of mutations selected from T85L, H121Y, N130K, M134L, E154D, N190D, N229E, V258G, A289P, N303S, and N336Q relative to SEQ ID NO: 35 increases the stability of the polypeptide, reduces or prevents binding of anti-drug antibodies to the polypeptide, eliminates T-cell binding epitopes, and eliminates chemical liability.In some embodiments, any one or set of mutations selected from T85L, H121Y, N130K, M134L, E154D, N190D, N229E, V258G, A289P, N303S, and N336Q relative to SEQ ID NO: 35 increases the stability of the polypeptide, reduces or prevents binding of anti-drug antibodies to the polypeptide, reduces or prevents T-cell binding to the polypeptide, and / or eliminates chemosensitivity. In some embodiments, a polypeptide comprising any one or set of mutations selected from T85L, H121Y, N130K, M134L, E154D, N190D, N229E, V258G, A289P, N303S, and N336Q relative to SEQ ID NO: 35 increases the stability, reduces or eliminates binding to anti-drug antibodies, eliminates a T-cell binding epitope, and / or eliminates chemosensitivity. In some embodiments, a polypeptide comprising any one or any set of mutations selected from T85L, H121Y, N130K, M134L, E154D, N190D, N229E, V258G, A289P, N303S, and N336Q relative to SEQ ID NO: 35 enhances stability, reduces or eliminates binding to anti-drug antibodies, does not bind to T cells, and / or does not have chemosensitivity. In some embodiments, any one or set of mutations selected from T85L, H121Y, M134L, E154D, N190D, N229E, V258G, A289P, N303S, and N336Q relative to SEQ ID NO: 35 enhances polypeptide stability, reduces or prevents binding of anti-drug antibodies to the polypeptide, eliminates a T-cell binding epitope, and eliminates chemosensitivity. In some embodiments, any one or set of mutations selected from T85L, H121Y, M134L, E154D, N190D, N229E, V258G, A289P, N303S, and N336Q relative to SEQ ID NO: 35 increases the stability of the polypeptide, reduces or prevents binding of anti-drug antibodies to the polypeptide, reduces or prevents binding of T cells to the polypeptide, and / or eliminates chemical liability.In some embodiments, a polypeptide comprising any one or any set of mutations selected from T85L, H121Y, M134L, E154D, N190D, N229E, V258G, A289P, N303S, and N336Q relative to SEQ ID NO: 35 has increased stability, reduced or no binding to anti-drug antibodies, no T-cell binding epitopes, and / or no chemosensitivity. In some embodiments, a polypeptide comprising any one or any set of mutations selected from T85L, H121Y, M134L, E154D, N190D, N229E, V258G, A289P, N303S, and N336Q relative to SEQ ID NO: 35 has increased stability, reduced or no binding to anti-drug antibodies, no T-cell binding epitopes, and / or no chemosensitivity. [Table 7-1] [Table 7-2] [Table 7-3] [Table 7-4] [Table 7-5] [Table 7-6] [Table 7-7] [Table 7-8] [Table 7-9] [Table 7-10] [Table 7-11] Table 7-12 Table 7-13 Table 7-14 Table 7-15 Table 7-16 Table 7-17 Table 7-18 Table 7-19 Table 7-20 Table 7-21 Table 7-22 Table 7-23 Table 7-24 Table 7-25 Table 7-26 Table 7-27 Table 7-28 Table 7-29 Table 7-30 Table 7-31 Table 7-32 Table 7-33 Table 7-34 Table 7-35 Table 7-36 Table 7-37 Table 7-38 Table 7-39 Table 7-40 Table 7-41 Table 7-42 Table 7-43 Table 7-44 Table 7-45 Table 7-46 Table 7-47 Table 7-48 Table 7-49 Table 7-50 Table 7-51 Table 7-52 Table 7-53 Table 7-54 Table 7-55 Table 7-56 Table 7-57 Table 7-58 Table 7-59 Table 7-60 Table 7-61 Table 7-62 Table 7-63

[0100] In some embodiments, the polypeptide is 50, 60, 70, 80, 85, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% identical to a reference protease (SEQ ID NOs: 35, 55-1019, 1290-1310, and 1333, 1023-1289, 1311-1329, or 1334-1336) and comprises a set of mutations described herein, such as those in the table above or in the following groups: In some embodiments, in addition to a set of mutations from such a reference sequence, the variant protease comprises a cysteine ​​(C) at a position in the variant sequence corresponding to position 94 of SEQ ID NO: 35, and / or has a lysine (K), a histidine (H), an aspartic acid (D), and / or an aspartic acid (D) at positions in the variant sequence corresponding to positions 84, 262, 284, and 286 of SEQ ID NO: 35, respectively. In some embodiments, the polypeptide comprises a lysine (K) at position 115 compared to SEQ ID NO:35. In some embodiments, the polypeptide comprises, or further comprises, an asparagine (N) at position 130 compared to SEQ ID NO:35. In some embodiments, the polypeptide comprises an asparagine (N) at position 336 compared to SEQ ID NO:35. In some embodiments, the polypeptide comprises a glutamine (Q) at position 337 compared to SEQ ID NO:35. In some embodiments, the polypeptide comprises a threonine (T) at position 338 compared to SEQ ID NO:35. In some embodiments, the polypeptide comprises an asparagine (N) at position 339 compared to SEQ ID NO:35. In some embodiments, the polypeptide comprises a glycine (G) at position 130 compared to SEQ ID NO:35. In some embodiments, the polypeptide comprises an asparagine (N) at position 284 compared to SEQ ID NO:35. In some embodiments, the polypeptide comprises an asparagine (N) at position 286 compared to SEQ ID NO:35. In some embodiments, the polypeptide comprises a lysine (K) at position 130 compared to SEQ ID NO:35. In some embodiments, the polypeptide comprises a glycine (G) at position 261 compared to SEQ ID NO: 35. In some embodiments, the polypeptide comprises a glycine (G) at position 258 compared to SEQ ID NO: 35.

[0101] In some embodiments, the polypeptide comprises any one or any combination of K115K, N130N, N336N, Q337Q, T338T, N339N, N130G, D284N, D286N, N130K, N261G, and V258G compared to SEQ ID NO: 35. In some embodiments, the polypeptide comprises any one of the set: K115K, N130N, N336N, Q337Q, T338T, and N339N compared to SEQ ID NO: 35; K115K, N130G, N336N, Q337Q, T338T, and N339N compared to SEQ ID NO: 35; D284N compared to SEQ ID NO: 35; D286N compared to SEQ ID NO: 35; N130K and D284N compared to SEQ ID NO: 35; N130K and D286N compared to SEQ ID NO: 35; K115K, N130N, N336N, Q337Q, T338T, N339N and N261G compared to SEQ ID NO: 35; K115K, N130G, N336N, Q337Q, T338T, N339N and N261G compared to SEQ ID NO: 35; D284N and and N261G, D286N and N261G compared to SEQ ID NO: 35, N130K, D284N and N261G compared to SEQ ID NO: 35, N130K, D286N and N261G compared to SEQ ID NO: 35, K115K, N130N, N336N, Q337Q, T338T, N339N and V258G compared to SEQ ID NO: 35, K115K, N130G, Includes N336N, Q337Q, T338T, N339N and V258G, D284N and V258G compared to SEQ ID NO: 35, D286N and V258G compared to SEQ ID NO: 35, N130K, D284N and V258G compared to SEQ ID NO: 35, N130K, D286N, V258G compared to SEQ ID NO: 35, or N130K and N261G compared to SEQ ID NO: 35.

[0102] In some embodiments, the protease comprises a lysine (K) at position 115 relative to SEQ ID NO:35. In some embodiments, the protease comprises an asparagine (N) at position 130 relative to SEQ ID NO:35. In some embodiments, the protease comprises an asparagine (N) at position 336 relative to SEQ ID NO:35. In some embodiments, the protease comprises a glutamine (Q) at position 337 relative to SEQ ID NO:35. In some embodiments, the protease comprises a threonine (T) at position 338 relative to SEQ ID NO:35. In some embodiments, the protease comprises an asparagine (N) at position 339 relative to SEQ ID NO:35. In some embodiments, the protease comprises a glycine (G) at position 130 relative to SEQ ID NO:35. In some embodiments, the protease comprises an asparagine (N) at position 284 relative to SEQ ID NO:35. In some embodiments, the protease comprises an asparagine (N) at position 286 relative to SEQ ID NO:35. In some embodiments, the protease comprises a lysine (K) at position 130 compared to SEQ ID NO: 35. In some embodiments, the protease comprises a glycine (G) at position 261 compared to SEQ ID NO: 35. In some embodiments, the protease comprises a glycine (G) at position 258 compared to SEQ ID NO: 35.

[0103] In some embodiments, the protease comprises any one or any combination of K115K, N130N, N336N, Q337Q, T338T, N339N, N130G, D284N, D286N, N130K, N261G, and V258G compared to SEQ ID NO: 35. In some embodiments, the protease comprises any one or any combination of K115K, N130N, N336N, Q337Q, T338T, and N339N compared to SEQ ID NO: 35, K115K, N130G, N336N, Q337Q, T338T, and N339N compared to SEQ ID NO: 35, D284N compared to SEQ ID NO: 35, D286N compared to SEQ ID NO: 35, N130K compared to SEQ ID NO: 35 K and D284N, N130K and D286N compared to SEQ ID NO: 35, K115K, N130N, N336N, Q337Q, T338T, N339N and N261G compared to SEQ ID NO: 35, K115K, N130G, N336N, Q337Q, T338T, N339N and N261G compared to SEQ ID NO: 35, D284N and N261G compared to SEQ ID NO: 35, SEQ ID NO: D286N and N261G compared to SEQ ID NO:35, N130K, D284N and N261G compared to SEQ ID NO:35, N130K, D286N and N261G compared to SEQ ID NO:35, K115K, N130N, N336N, Q337Q, T338T, N339N and V258G compared to SEQ ID NO:35, K115K, N130G, N336N, Q337Q , T338T, N339N and V258G, D284N and V258G compared to SEQ ID NO: 35, D286N and V258G compared to SEQ ID NO: 35, N130K, D284N and V258G compared to SEQ ID NO: 35, N130K, D286N, V258G compared to SEQ ID NO: 35, or N130K and N261G compared to SEQ ID NO: 35.

[0104] In some embodiments, the polypeptide comprises, or further comprises, a mutation at any one position, or any combination of positions, selected from 130, 131, 198, 216, 302, 119, 244, 142, 115, 241, 245, 316, 333, 139, relative to SEQ ID NO: 35. In some embodiments, the polypeptide comprises a positively charged amino acid at position 130, relative to SEQ ID NO: 35, optionally, the positively charged amino acid is arginine (R) or lysine (K). In some embodiments, the polypeptide comprises a positively charged amino acid at position 131, relative to SEQ ID NO: 35, optionally, the positively charged amino acid is arginine (R) or lysine (K). In some embodiments, the polypeptide comprises a positively charged amino acid at position 130, relative to SEQ ID NO: 35, optionally, the positively charged amino acid is arginine (R) or lysine (K). The polypeptide comprises a positively charged amino acid at position 131 compared to SEQ ID NO: 35, and optionally, the positively charged amino acid is arginine (R) or lysine (K).

[0105] In some embodiments, the polypeptide comprises any one mutation, or any set of mutations, selected from N130K, E198R, D216N, N130R, E198K, S302K, E119R, T244D, E119K, D142R, T244E, K115E, K241E, E245K, D316K, D333K, K241S, E245N, E139K. In some embodiments, the polypeptides are N130K, E198R and D216N, N130R, E198K, D216N and S302K, E119R, D216N and T244D, E119K, D142R, D216N, T244E and S302K, K115E, D216N, K241E, E245K, D316K and D333K , E119K, N130R, D142R, D216N, K241S, T244E, E245N and S302K, E119K, N130R, D142R, E198K, D216N and T244E, and K115E, N130R, E198K, D216N, K241E, E245K and D333K.

[0106] In some embodiments, the polypeptide comprises a mutation at any one position or any combination of positions selected from 84, 93, 95, 97, 137, 140, 147, 150, 162, 165, 166, 171, 174, 205, 226, 237, 239, 243, 250, 251, 254, 255, 282, 288, 312, 315, 347, 349 relative to SEQ ID NO: 1331 or 1332. In some embodiments, the polypeptide comprises a positively charged amino acid at position 138 relative to SEQ ID NO: 1331 or 1332, optionally wherein the positively charged amino acid is arginine (R) or lysine (K). In some embodiments, the polypeptide comprises a positively charged amino acid at position 139 compared to SEQ ID NO: 1331 or 1332, and optionally, the positively charged amino acid is arginine (R) or lysine (K). In some embodiments, the polypeptide comprises a positively charged amino acid at position 138 compared to SEQ ID NO: 1331 or 1332, and optionally, the positively charged amino acid is arginine (R) or lysine (K), and the polypeptide comprises a positively charged amino acid at position 139 compared to SEQ ID NO: 1331 or 1332, and optionally, the positively charged amino acid is arginine (R) or lysine (K).

[0107] In some embodiments, the polypeptide comprises any one mutation, or any set of mutations, selected from H84N, A93T, D95N, K97A, F137I, Q140E, A147E, D150R, N162E, R165K, D166E, N171Y, A174T, N205K, D226N, L237F, N239E, N243K, K250S, Q251E, T254E, E255K, N282D, E288K, A312K, H315K, K347Q, S349N. In some embodiments, the polypeptide comprises H84N, N138R, A147E, D150R, N162E, N171Y, N205K, D226N, Q251E, E255K, A312K and S349N, A93T, D95N, Q140E, R165K, D166E, A174T, D226N, L237F, N239E, N243K, N282D, E288K, H315K and K347 and any set of mutations selected from Q, R70T, N72Q and N73G, R70T, N72Q, N73G, H84N, N138R, N162E, N205K and D226N, R70T, N72Q, N73G, N138R and D226N, R70T, N72Q, N73G, K97A, N138R and D226N, and R70T, N72Q, N73G, N138R and D226N.

[0108] In some embodiments, the protease comprises a mutation at any one position, or any combination of positions, selected from 130, 131, 198, 216, 302, 119, 244, 142, 115, 241, 245, 316, 333, 139 relative to SEQ ID NO: 35. In some embodiments, the protease comprises a positively charged amino acid at position 130 relative to SEQ ID NO: 35, optionally wherein the positively charged amino acid is arginine (R) or lysine (K). In some embodiments, the protease comprises a positively charged amino acid at position 131 relative to SEQ ID NO: 35, optionally wherein the positively charged amino acid is arginine (R) or lysine (K). In some embodiments, the protease comprises a positively charged amino acid at position 130 compared to SEQ ID NO: 35, optionally the positively charged amino acid is arginine (R) or lysine (K), and the protease comprises a positively charged amino acid at position 131 compared to SEQ ID NO: 35, optionally the positively charged amino acid is arginine (R) or lysine (K).

[0109] In some embodiments, the protease comprises any one mutation or set of mutations selected from N130K, E198R, D216N, N130R, E198K, S302K, E119R, T244D, E119K, D142R, T244E, K115E, K241E, E245K, D316K, D333K, K241S, E245N, E139K. In some embodiments, the proteases are N130K, E198R and D216N, N130R, E198K, D216N and S302K, E119R, D216N and T244D, E119K, D142R, D216N, T244E and S302K, K115E, D216N, K241E, E245K, D316K and D333K , E119K, N130R, D142R, D216N, K241S, T244E, E245N and S302K, E119K, N130R, D142R, E198K, D216N and T244E, and K115E, N130R, E198K, D216N, K241E, E245K and D333K.

[0110] In some embodiments, the protease comprises a mutation at any one position or any combination of positions selected from 84, 93, 95, 97, 137, 140, 147, 150, 162, 165, 166, 171, 174, 205, 226, 237, 239, 243, 250, 251, 254, 255, 282, 288, 312, 315, 347, 349 relative to SEQ ID NO: 1331 or 1332. In some embodiments, the protease comprises a positively charged amino acid at position 138 relative to SEQ ID NO: 1331 or 1332, optionally wherein the positively charged amino acid is arginine (R) or lysine (K). In some embodiments, the protease comprises a positively charged amino acid at position 139 compared to SEQ ID NO: 1331 or 1332, and optionally the positively charged amino acid is arginine (R) or lysine (K). In some embodiments, the protease comprises a positively charged amino acid at position 138 compared to SEQ ID NO: 1331 or 1332, and optionally the positively charged amino acid is arginine (R) or lysine (K), and the protease comprises a positively charged amino acid at position 139 compared to SEQ ID NO: 1331 or 1332, and optionally the positively charged amino acid is arginine (R) or lysine (K).

[0111] In some embodiments, the protease comprises any one mutation or set of mutations selected from H84N, A93T, D95N, K97A, F137I, Q140E, A147E, D150R, N162E, R165K, D166E, N171Y, A174T, N205K, D226N, L237F, N239E, N243K, K250S, Q251E, T254E, E255K, N282D, E288K, A312K, H315K, K347Q, S349N. In some embodiments, the protease comprises any set of mutations selected from H84N, N138R, A147E, D150R, N162E, N171Y, N205K, D226N, Q251E, E255K, A312K, and S349N. and any set of mutations selected from A93T, D95N, Q140E, R165K, D166E, A174T, D226N, L237F, N239E, N243K, N282D, E288K, H315K and K347Q, R70T, N72Q and N73G, R70T, N72Q, N73G, H84N, N138R, N162E, N205K and D226N, R70T, N72Q, N73G, N138R and D226N, R70T, N72Q, N73G, K97A, N138R and D226N, and R70T, N72Q, N73G, N138R and D226N.

[0112] In some embodiments, the polypeptide comprises any mutation or set of mutations provided in PCT Publication Nos. WO2016128558 and WO2016128559, or U.S. Pat. Nos. 10,696,959, 11,214,784, 10,758,597, or 11,524,057, each of which is incorporated by reference in its entirety.

[0113] IdeS (Streptococcus pyogenes immunoglobulin G-degrading enzyme), an enzyme derived from Streptococcus pyogenes that cleaves IgG antibodies, is being investigated for its potential therapeutic applications, particularly in the field of transplant immunology. The immunogenicity of IdeS is an important consideration for its clinical use, as the human immune system could potentially recognize the enzyme as a foreign protein and trigger an immune response. This response could lead to the production of anti-drug antibodies (ADAs), which could neutralize IdeS's enzymatic activity or alter its pharmacokinetics, potentially reducing its efficacy or increasing the risk of side effects. Furthermore, repeated administration of IdeS could intensify this immunogenic response, limiting its long-term usefulness in chronic conditions. Therefore, for any given subject, issues related to IdeS's immunogenicity are likely to present a barrier to its use as a treatment. These issues could necessitate increased doses of IdeS and / or preclude treatment with IdeS altogether, especially if repeated administration is required.

[0114] Existing approaches to this type of problem include, for example, pegylation of therapeutic agents to reduce immunogenicity or coadministration of therapeutic agents with immunosuppressants. Other approaches include identifying specific positions within the sequence of IdeS that, when modified, result in polypeptides that alleviate immunogenicity-related problems compared to IdeS. Thus, some modifications may indirectly reduce immunogenicity by increasing the effectiveness of the polypeptides of the present disclosure in cleaving IgG compared to IdeS, thereby allowing the use of lower doses or concentrations 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 polypeptides of the present disclosure compared to IdeS.

[0115] The present disclosure provides IdeS variants that, among other properties, have reduced immunogenicity compared to wild-type IdeS, allowing for multiple administrations of the variants to patients. The variants provided herein can also contain mutations or changes, compared to the wild-type sequence, that affect post-translational modifications, aggregation, glycosylation, impurities, and formulation components; reduce B-cell epitopes, thereby reducing the likelihood of forming antibody epitopes that generate antibodies against the variant in vivo; reduce T-cell epitopes (linear peptide sequences that can be displayed on MHC II and elicit a novel immune response); and reduce the likelihood of ADA (anti-drug antibodies). Thus, in some embodiments, a polypeptide having IgG protease activity has one or more B-cell epitopes ablated. In some embodiments, a polypeptide having IgG protease activity has one or more T-cell epitopes ablated. In some embodiments, a polypeptide having IgG protease activity has no significant chemical liability. In some embodiments, the polypeptides having IgG protease activity have one or more B cell epitopes removed, one T cell epitope removed, and no significant chemical liability. Without being bound by any particular theory, polypeptides having IgG protease activity, such as those provided herein, can be utilized in a multi-administration manner and, therefore, can be used in a non-dose-limiting manner or manner.

[0116] In some embodiments, a polypeptide having IgG protease activity is as effective or more effective at cleaving IgG as a wild-type protease and / or is less immunogenic than a wild-type protease. In some embodiments, a polypeptide having IgG protease activity is as effective at cleaving IgG as a wild-type protease. In some embodiments, a polypeptide having IgG protease activity is more effective at cleaving IgG than a wild-type protease. In some embodiments, a polypeptide having IgG protease activity is less effective at cleaving IgG than a wild-type protease, but is less immunogenic and / or less chemically labile than a wild-type protease. These properties can be compared to wild-type IdeS or wild-type IdeS fused to an Fc molecule, such as those provided herein. In some embodiments, a polypeptide having IgG protease activity is less immunogenic than a wild-type protease. In some embodiments, a polypeptide having IgG protease activity is as immunogenic as a wild-type protease.

[0117] Immunogenicity can be measured by any assay, such as that described in Example 15. For example, the immunogenicity of a variant fused or not to Fc can be compared to wild-type IdeS (fused or not to Fc) by assessing pre-existing antibodies binding to the variant in human donor cells, such as PBMCs, if the variant and wild-type IdeS molecules are the same type of molecule (fused or not to Fc), and can be measured by electrochemiluminescence (ECL) immunoassay. This is one exemplary method, and other methods can also be used.

[0118] In some embodiments, a polypeptide having IgG protease activity is as effective or more effective at cleaving IgG as wild-type IdeS and / or is less immunogenic than or similar to wild-type IdeS. In some embodiments, a polypeptide having IgG protease activity is as effective at cleaving IgG as wild-type IdeS. In some embodiments, a polypeptide having IgG protease activity is more effective at cleaving IgG than wild-type IdeS. In some embodiments, a polypeptide having IgG protease activity is less immunogenic than wild-type IdeS. In some embodiments, a polypeptide having IgG protease activity is similar to wild-type IdeS in immunogenicity.

[0119] In some embodiments, PRT-1 through PRT-719 are as effective or more effective at cleaving IgG as wild-type proteases and / or are less or equally immunogenic than wild-type proteases. In some embodiments, PRT-1 through PRT-719 are as effective at cleaving IgG as wild-type proteases. In some embodiments, PRT-1 through PRT-719 are more effective at cleaving IgG than wild-type proteases. In some embodiments, PRT-1 through PRT-719 are less immunogenic than wild-type proteases. In some embodiments, PRT-1 through PRT-719 are equally immunogenic as wild-type proteases.

[0120] In some embodiments, PRT-1 through PRT-719 are as effective or more effective at cleaving IgG as wild-type proteases and / or are less immunogenic than or similar to wild-type IdeS. In some embodiments, PRT-1 through PRT-719 are as effective at cleaving IgG as wild-type IdeS. In some embodiments, PRT-1 through PRT-719 are more effective at cleaving IgG than wild-type IdeS. In some embodiments, PRT-1 through PRT-719 are less immunogenic than wild-type IdeS. In some embodiments, PRT-1 through PRT-719 are similarly immunogenic to wild-type IdeS.

[0121] In some embodiments, VRT-1 through VRT-319 are as effective or more effective at cleaving IgG as wild-type proteases and / or are less or equally immunogenic than wild-type proteases. In some embodiments, VRT-1 through VRT-319 are as effective at cleaving IgG as wild-type proteases. In some embodiments, VRT-1 through VRT-319 are more effective at cleaving IgG than wild-type proteases. In some embodiments, VRT-1 through VRT-319 are less immunogenic than wild-type proteases. In some embodiments, VRT-1 through VRT-319 are equally immunogenic as wild-type proteases.

[0122] In some embodiments, VRT-1 through VRT-319 are as effective or more effective at cleaving IgG as wild-type proteases and / or are less immunogenic than or similar to wild-type IdeS. In some embodiments, VRT-1 through VRT-319 are as effective at cleaving IgG as wild-type IdeS. In some embodiments, VRT-1 through VRT-319 are more effective at cleaving IgG than wild-type IdeS. In some embodiments, VRT-1 through VRT-319 are less immunogenic than wild-type IdeS. In some embodiments, VRT-1 through VRT-319 are similarly immunogenic to wild-type IdeS.

[0123] Fc fusion molecule Provided herein are therapeutic compounds, e.g., therapeutic protein molecules, e.g., fusion proteins, comprising 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 variant Fc polypeptide, such as, but not limited to, those provided herein.

[0124] The present disclosure provides, for example, a polypeptide comprising a polypeptide having protease activity and an Fc polypeptide domain, wherein the polypeptide having protease activity is covalently or non-covalently linked to the Fc polypeptide domain. In some embodiments, the Fc polypeptide domain is a variant Fc polypeptide, such as those provided herein. Thus, in some embodiments, a polypeptide comprising a polypeptide having protease activity and an Fc polypeptide domain is provided, wherein the polypeptide having protease activity is covalently or non-covalently linked to the Fc polypeptide. 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.

[0125] In some embodiments, the Fc polypeptide domain is an IgG Fc polypeptide, such as an Fc polypeptide that is or is derived from an IgG1, IgG2, IgG3, or IgG4 Fc polypeptide. In some embodiments, the Fc polypeptide domain is a mutant Fc polypeptide domain, such as the variants provided herein. In some embodiments, the Fc polypeptide domain is protease resistant. An Fc polypeptide is resistant to the protease to which it is attached and can therefore be made resistant to protease cleavage and / or binding by a protease. In some embodiments, a mutant 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, a mutant 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, a mutant 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, variant Fc polypeptides such as those provided herein comprise a set of mutations that render the IgG Fc polypeptide resistant to binding by a protease or variants thereof. In some embodiments, variant Fc polypeptides such as those provided herein comprise mutations that render the IgG Fc polypeptide resistant to cleavage by a protease or variants thereof and / or binding by a protease or variants thereof. In some embodiments, variant Fc polypeptides such as those provided herein comprise a set of mutations that render the IgG Fc polypeptide resistant to cleavage by a protease or variants thereof and / or binding by a protease or variants thereof. In some embodiments, variant Fc polypeptides such as those provided herein are resistant to cleavage by a protease or variants thereof. In some embodiments, variant Fc polypeptides such as those provided herein are resistant to binding by a protease or variants thereof.In some embodiments, variant Fc polypeptides such as those provided herein are resistant to cleavage by a protease or variant thereof and / or binding by a protease or variant thereof. Variant Fc polypeptides can also be made resistant to other proteases to which the Fc polypeptide domain is not attached. Fc polypeptides can self-associate with each other to form dimers. Dimers can be homodimers, in which the Fc polypeptide is the same in each polypeptide molecule, or heterodimers, in which the Fc polypeptide is different in each polypeptide molecule that forms the dimer.

[0126] In some embodiments, the protease-resistant Fc domain is resistant to cleavage by IdeS protease or a variant thereof. In some embodiments, the protease-resistant Fc domain is completely resistant to cleavage by IdeS protease or a variant thereof. In some embodiments, the protease-resistant Fc domain is about 10%, about 20%, about 30%, about 40%, about 50%, about 60%, about 70%, about 80%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99%, or about 100% resistant to cleavage by IdeS protease or a variant thereof. In some embodiments, the protease-resistant Fc domain is completely resistant to cleavage by IdeS protease or a variant thereof. In some embodiments, the protease-resistant Fc domain is 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% resistant to cleavage by IdeS protease or a variant thereof. In some embodiments, the protease-resistant Fc domain is more resistant to cleavage by IdeS protease or a variant thereof compared to a wild-type Fc domain. In some embodiments, the protease-resistant Fc domain is completely resistant to cleavage by IdeS protease or a variant thereof compared to a wild-type Fc domain. In some embodiments, a protease-resistant Fc domain is about 10%, about 20%, about 30%, about 40%, about 50%, about 60%, about 70%, about 80%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99%, or about 100% resistant to cleavage by IdeS protease or a variant thereof compared to a wild-type Fc domain. In some embodiments, a protease-resistant Fc domain is completely resistant to cleavage by IdeS protease or a variant thereof.In some embodiments, the protease-resistant Fc domain is 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% resistant to cleavage by the IdeS protease or a variant thereof compared to a wild-type Fc domain. In some embodiments, the protease-resistant Fc domain is resistant to cleavage by the IdeS protease or a variant thereof compared to an Fc domain that does not comprise any one or all of the mutations in SEQ ID NO: 23. In some embodiments, the protease-resistant Fc domain is completely resistant to cleavage by the IdeS protease or a variant thereof compared to an Fc domain that does not comprise any one or all of the mutations in SEQ ID NO: 23. In some embodiments, the protease-resistant Fc domain is about 10%, about 20%, about 30%, about 40%, about 50%, about 60%, about 70%, about 80%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99%, or about 100% resistant to cleavage by the IdeS protease or a variant thereof compared to an Fc domain that does not contain any one or all of the mutations of SEQ ID NO: 23. In some embodiments, the protease-resistant Fc domain is completely resistant to cleavage by the IdeS protease or a variant thereof. In some embodiments, the protease-resistant Fc domain is 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% resistant to cleavage by the IdeS protease or a variant thereof compared to an Fc domain that does not contain any one or all of the mutations of SEQ ID NO:23.

[0127] In some embodiments, a variant Fc polypeptide is covalently or non-covalently conjugated to a polypeptide having IgG protease activity, and the polypeptide having IgG protease activity can be conjugated to the N-terminus or C-terminus of the variant Fc polypeptide. In some embodiments, a variant Fc polypeptide is covalently or non-covalently conjugated to a polypeptide having IgG protease activity, and the polypeptide having IgG protease activity can be conjugated to the N-terminus of the variant Fc polypeptide. In some embodiments, a variant Fc polypeptide is covalently or non-covalently conjugated to a polypeptide having IgG protease activity, and the polypeptide having IgG protease activity can be conjugated to the C-terminus of the variant Fc polypeptide.

[0128] The Fc polypeptide and the polypeptide having protease activity can be physically tethered to each other, covalently or non-covalently, directly or via a linker, e.g., as domains of the same linear primary amino acid sequence within a single polypeptide. The polypeptides can associate with each other via the Fc polypeptide such that a dimer having a first polypeptide and a second polypeptide is generated. In some embodiments, the first polypeptide comprises a polypeptide having protease activity linked to an Fc polypeptide domain. In some embodiments, the second polypeptide comprises an Fc polypeptide domain, such as those provided herein. In some embodiments, the second polypeptide does not comprise a protease domain. In some embodiments, the second polypeptide comprises a protease domain.

[0129] This multidomain 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, such as a fusion protein.

[0130] In some embodiments, the polypeptide comprising the polypeptide with protease activity, the Fc polypeptide domain, which may be a variant Fc polypeptide, or both, further comprises a single-domain antibody molecule, e.g., a nanobody, a camelid antibody VHH molecule, or a human soluble VH domain. Without wishing to be bound by any particular theory, a nanobody covalently or non-covalently conjugated to a polypeptide with protease activity and / or a polypeptide comprising an Fc polypeptide may extend the half-life of the polypeptide. In some embodiments, the single-domain antibody molecule, e.g., a nanobody, a camelid antibody VHH molecule, or a human soluble VH domain, is physically tethered, covalently or non-covalently, directly or via a linker, to the polypeptide with protease activity, the variant Fc polypeptide, or both. In some embodiments, the variant Fc polypeptide may also comprise a single-chain variable fragment (scFv) or a Fab domain. In some embodiments, a therapeutic protein molecule, or a nucleic acid (e.g., mRNA or DNA) encoding a therapeutic protein molecule, can be administered to a subject. In some embodiments, the polypeptide with protease activity and the variant Fc polypeptide are linked to a third entity, such as a carrier (e.g., a polymeric carrier, a dendrimer), or a particle (e.g., a nanoparticle). As used herein, the terms "nanobody" and "VHH" can be used interchangeably.

[0131] Non-limiting examples of nanobodies include those provided in McMahon et al., Nat Struct Mol Biol. 2018 Mar;25(3):289-296. doi:10.1038 / s41594-018-0028-6, which is incorporated by reference in its entirety. In some embodiments, the nanobody or VHH comprises an amino acid sequence set forth in Table 6. [Table 8]

[0132] In some embodiments, a Nanobody 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%, at least 99%, or 100% sequence identity to any of the sequences provided in Table 6. In some embodiments, a Nanobody 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: 30, 32, or 33. In some embodiments, a Nanobody comprises an amino acid sequence that has 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: 30. In some embodiments, a Nanobody comprises an amino acid sequence that has 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: 32. In some embodiments, the Nanobody 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: 33.

[0133] In some embodiments, a Nanobody comprises an amino acid sequence having the amino acid sequence of SEQ ID NO: 30, 32, or 33. In some embodiments, a Nanobody comprises an amino acid sequence having the amino acid sequence of SEQ ID NO: 30. In some embodiments, a Nanobody comprises an amino acid sequence having the amino acid sequence of SEQ ID NO: 32. In some embodiments, a Nanobody comprises an amino acid sequence having the amino acid sequence of SEQ ID NO: 33.

[0134] In some embodiments, a therapeutic compound or compounds comprise a polypeptide with protease activity covalently or non-covalently conjugated to a variant Fc polypeptide. In some embodiments, a therapeutic molecule comprises a fusion protein comprising a polypeptide with protease activity fused to a variant Fc polypeptide, e.g., directly or via a linking moiety comprising one or more amino acid residues. In some embodiments, a therapeutic compound or compounds comprise a polypeptide comprising a polypeptide with protease activity linked to a variant Fc polypeptide non-covalently or covalently, e.g., by a covalent bond other than a peptide bond, e.g., a sulfhydryl bond.

[0135] 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 any of the variant proteases described above. Examples of IgG proteases and variant proteases include WO2003051914, WO2006131347, WO2016128558, WO2016128559, WO2016012285, WO2008071418, WO2012119983, WO2010057626, WO2015184325, WO2021026264, WO2021021989, WO2018034346, WO2010089126, WO200908027, WO2008136735, WO2015 No. 181356, WO2018093868, WO2013037824, WO2017134274, WO2016046220, WO2015040125, WO2009033670, WO2004096157, WO2010123885, WO2010118337, WO2019075360, WO2007019376, and U.S. Pat. No. 10,836,815, each of which is incorporated herein by reference in its entirety.Non-limiting examples of proteases include, but are not limited to, proteases comprising an amino acid sequence that has 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, Table 5, Table 7, or Table 8.

[0136] In some embodiments, the polypeptide comprises a polypeptide with protease activity covalently or non-covalently conjugated to the N-terminus of a variant Fc polypeptide. In some embodiments, the polypeptide comprises a polypeptide with protease activity covalently or non-covalently conjugated to the C-terminus of a variant Fc polypeptide. In some embodiments, a linker moiety, such as a peptide moiety or a non-peptide bond, can be present between the polypeptide with protease activity and the N-terminus or C-terminus of the Fc polypeptide.

[0137] In some embodiments, the polypeptide comprises a polypeptide with protease activity and a variant Fc polypeptide covalently or non-covalently conjugated to a Nanobody, wherein the C-terminus of the polypeptide with protease activity is covalently or non-covalently conjugated to the N-terminus of the Nanobody and the N-terminus of the polypeptide with protease activity is covalently or non-covalently conjugated to the C-terminus of the variant Fc polypeptide. In some embodiments, the polypeptide comprises a polypeptide with protease activity and a variant Fc polypeptide covalently or non-covalently conjugated to a Nanobody, wherein the N-terminus of the polypeptide with protease activity is covalently or non-covalently conjugated to the C-terminus of the Nanobody and the C-terminus of the polypeptide with protease activity is covalently or non-covalently conjugated to the N-terminus of the variant Fc polypeptide. In some embodiments, the polypeptide with protease activity is as provided herein. In some embodiments, the variant IgG Fc is as provided herein. In some embodiments, the Nanobody is as provided herein.

[0138] In some embodiments, a polypeptide comprising a polypeptide with protease activity covalently or non-covalently conjugated to a variant Fc polypeptide further comprises a tag, such as a purification tag or a detection tag. This tag can be used to facilitate purification, isolation, or detection of the polypeptide. A non-limiting example of such a tag is a histidine tag, which is a plurality of histidines (e.g., six or more histidines). In some embodiments, a polypeptide comprising a polypeptide with protease activity covalently or non-covalently conjugated to a variant Fc polypeptide further comprises a histidine tag, wherein the C-terminus of the polypeptide with protease activity is covalently or non-covalently conjugated to the N-terminus of the variant Fc polypeptide, and the C-terminus of the variant Fc polypeptide is covalently or non-covalently conjugated to the N-terminus of the tag. In some embodiments, a polypeptide comprising a polypeptide with protease activity and a variant Fc polypeptide covalently or non-covalently conjugated to a Nanobody further comprises a tag. In some embodiments, the polypeptide comprising a polypeptide with protease activity and a variant Fc polypeptide covalently or non-covalently conjugated to a Nanobody further comprises a tag, wherein the N-terminus of the polypeptide with protease activity is covalently or non-covalently conjugated to the C-terminus of the Nanobody, the C-terminus of the polypeptide with protease activity is covalently or non-covalently conjugated to the N-terminus of the variant Fc polypeptide, and the C-terminus of the variant Fc polypeptide is covalently or non-covalently conjugated to the N-terminus of the tag.In some embodiments, the polypeptide comprising a polypeptide with protease activity and a variant Fc polypeptide covalently or non-covalently conjugated to a Nanobody further comprises a histidine tag, wherein the N-terminus of the polypeptide with protease activity is covalently or non-covalently conjugated to the C-terminus of the variant Fc polypeptide, the C-terminus of the polypeptide with protease activity is covalently or non-covalently conjugated to the N-terminus of the Nanobody, and the C-terminus of the Nanobody is covalently or non-covalently conjugated to the N-terminus of the tag.

[0139] In some embodiments, the 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 a sequence provided in Table 3. In some embodiments, the polypeptide comprises a polypeptide having protease activity covalently or non-covalently conjugated to a variant Fc polypeptide, wherein the variant Fc polypeptide comprises an amino acid sequence that has 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: 19, 20, 21, 22, 23, 24, 25, 42, 43, 44, 45, 46, 47, 48, or 49. In some embodiments, the polypeptide comprises a polypeptide having protease activity covalently or non-covalently conjugated to a variant Fc polypeptide, wherein the variant Fc polypeptide comprises an amino acid sequence that has 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.

[0140] In some embodiments, the polypeptide comprises a polypeptide having protease activity covalently or non-covalently conjugated to a variant Fc polypeptide, wherein the 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 a sequence provided in Table 3, and further comprises one or more mutations at positions 234, 235, 237, 296, 329, or any combination thereof.

[0141] In some embodiments, the polypeptide comprises a polypeptide having protease activity covalently or non-covalently conjugated to a variant Fc polypeptide, wherein the 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 a sequence provided in Table 3, and further comprises one or more mutations at positions L234, L235, G237, Y296, P329, or any combination thereof.

[0142] As used herein, the positions in an Fc polypeptide referred to as positions of mutations refer to the EU numbering system. The Fc polypeptide can be aligned with the wild-type sequence to determine the positions that are mutated according to the numbering system.

[0143] As used herein with reference to a polypeptide having a % identity to a reference sequence and further comprising mutations at one or more positions, means a polypeptide having the recited % identity to the reference sequence and also having one or more mutations at the recited positions.

[0144] In some embodiments, the polypeptide comprises a polypeptide having protease activity covalently or non-covalently conjugated to a variant Fc polypeptide, wherein the 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, and further comprises one or more mutations at positions 234, 235, 237, 296, 329, or any combination thereof.

[0145] In some embodiments, the polypeptide comprises a polypeptide having protease activity covalently or non-covalently conjugated to a variant Fc polypeptide, wherein the 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, and further comprises one or more mutations at positions 234, 235, 237, 296, 329, or any combination thereof.

[0146] In some embodiments, the polypeptide comprises a polypeptide having protease activity covalently or non-covalently conjugated to a variant Fc polypeptide, wherein the 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, and further comprises one or more mutations at positions L234, L235, G237, Y296, P329, or any combination thereof.

[0147] In some embodiments, the polypeptide comprises a polypeptide having protease activity covalently or non-covalently conjugated to a variant Fc polypeptide, wherein the 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, and further comprises one or more mutations at positions L234, L235, G237, Y296, P329, or any combination thereof.

[0148] In some embodiments, the polypeptide comprises a polypeptide having protease activity covalently or non-covalently conjugated to a variant Fc polypeptide, wherein the 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 any of the sequences provided in Table 3, and further comprises one or more mutations: L234A, L235A, G237A, Y296Q, P329K, or any combination thereof.

[0149] In some embodiments, the polypeptide comprises a polypeptide having protease activity covalently or non-covalently conjugated to a variant Fc polypeptide, wherein the 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, and further comprises one or more mutations of L234A, L235A, G237A, Y296Q, P329K, or any combination thereof.

[0150] In some embodiments, the polypeptide comprises a polypeptide having protease activity covalently or non-covalently conjugated to a variant Fc polypeptide, wherein the 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, and further comprises one or more mutations: L234A, L235A, G237A, Y296Q, P329K, or any combination thereof.

[0151] In some embodiments, the polypeptide comprises a polypeptide with protease activity covalently or non-covalently conjugated to a variant Fc polypeptide, wherein the variant Fc polypeptide comprises an amino acid sequence provided in Table 3.

[0152] In some embodiments, the polypeptide comprises a polypeptide with protease activity covalently or non-covalently conjugated to a variant Fc polypeptide, wherein the variant Fc polypeptide comprises the amino acid sequence of SEQ ID NO: 19, 20, 21, 22, 23, 24, 25, 42, 43, 44, 45, 47, 48, or 49. In some embodiments, the polypeptide comprises a polypeptide with protease activity covalently or non-covalently conjugated to a variant Fc polypeptide, wherein the variant Fc polypeptide comprises the amino acid sequence of SEQ ID NO: 19. In some embodiments, the polypeptide comprises a polypeptide with protease activity covalently or non-covalently conjugated to a variant Fc polypeptide, wherein the variant Fc polypeptide comprises the amino acid sequence of SEQ ID NO: 20. In some embodiments, the polypeptide comprises a polypeptide with protease activity covalently or non-covalently conjugated to a variant Fc polypeptide, wherein the variant Fc polypeptide comprises the amino acid sequence of SEQ ID NO: 21. In some embodiments, the polypeptide comprises a polypeptide with protease activity covalently or non-covalently conjugated to a variant Fc polypeptide, wherein the variant Fc polypeptide comprises the amino acid sequence of SEQ ID NO: 22. In some embodiments, the polypeptide comprises a polypeptide with protease activity covalently or non-covalently conjugated to a variant Fc polypeptide, wherein the variant Fc polypeptide comprises the amino acid sequence of SEQ ID NO: 23. In some embodiments, the polypeptide comprises a polypeptide with protease activity covalently or non-covalently conjugated to a variant Fc polypeptide, wherein the variant Fc polypeptide comprises the amino acid sequence of SEQ ID NO: 24. In some embodiments, the polypeptide comprises a polypeptide with protease activity covalently or non-covalently conjugated to a variant Fc polypeptide, wherein the variant Fc polypeptide comprises the amino acid sequence of SEQ ID NO: 25.In some embodiments, the polypeptide comprises a polypeptide with protease activity covalently or non-covalently conjugated to a variant Fc polypeptide, wherein the variant Fc polypeptide comprises the amino acid sequence of SEQ ID NO: 42. In some embodiments, the polypeptide comprises a polypeptide with protease activity covalently or non-covalently conjugated to a variant Fc polypeptide, wherein the variant Fc polypeptide comprises the amino acid sequence of SEQ ID NO: 43. In some embodiments, the polypeptide comprises a polypeptide with protease activity covalently or non-covalently conjugated to a variant Fc polypeptide, wherein the variant Fc polypeptide comprises the amino acid sequence of SEQ ID NO: 44. In some embodiments, the polypeptide comprises a polypeptide with protease activity covalently or non-covalently conjugated to a variant Fc polypeptide, wherein the variant Fc polypeptide comprises the amino acid sequence of SEQ ID NO: 45. In some embodiments, the polypeptide comprises a polypeptide with protease activity covalently or non-covalently conjugated to a variant Fc polypeptide, wherein the variant Fc polypeptide comprises the amino acid sequence of SEQ ID NO: 46. In some embodiments, the polypeptide comprises a polypeptide with protease activity covalently or non-covalently conjugated to a variant Fc polypeptide, wherein the variant Fc polypeptide comprises the amino acid sequence of SEQ ID NO: 47. In some embodiments, the polypeptide comprises a polypeptide with protease activity covalently or non-covalently conjugated to a variant Fc polypeptide, wherein the variant Fc polypeptide comprises the amino acid sequence of SEQ ID NO: 48. In some embodiments, the polypeptide comprises a polypeptide with protease activity covalently or non-covalently conjugated to a variant Fc polypeptide, wherein the variant Fc polypeptide comprises the amino acid sequence of SEQ ID NO: 49.

[0153] In some embodiments, the polypeptide comprises a polypeptide with protease activity covalently or non-covalently conjugated to a variant Fc polypeptide, wherein the variant Fc polypeptide comprises an amino acid sequence with a C-terminal lysine (K). In some embodiments, the polypeptide comprises a polypeptide with protease activity covalently or non-covalently conjugated to a variant Fc polypeptide, wherein the variant Fc polypeptide comprises an amino acid sequence that does not have (does not include) a C-terminal lysine (K).

[0154] In some embodiments, the protease present in the polypeptides provided herein has 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 or identity to a sequence provided in Table 4, Table 5, Table 7, or Table 8. and wherein the protease is covalently or non-covalently conjugated to the variant Fc polypeptide, wherein the 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 any of the sequences provided in Table 3.

[0155] In some embodiments, the polypeptide 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 NOs: 26, 28, 29, 31, 36, 37, 38, 39, 40, 41, 50, 51, 52, 55-1019, 1290-1310, or 1333; The polypeptide having ATPase activity is covalently or non-covalently conjugated to a variant Fc polypeptide, wherein the 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, 25, 42, 43, 44, 45, 47, 48, or 49.

[0156] In some embodiments, the polypeptide is a protease 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: 26, 28, 29, 31, 36, 37, 38, 39, 40, 41, 50, 51, 52, 55-1019, 1290-1310, or 1333. The polypeptide having protease activity is covalently or non-covalently conjugated to a variant Fc polypeptide, wherein the 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.

[0157] In some embodiments, the polypeptide comprises a polypeptide having protease activity covalently or non-covalently conjugated to a variant Fc polypeptide, wherein the 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 any of the sequences provided in Table 3, and further comprises one or more mutations at positions 234, 235, 237, 296, 329, or any combination thereof.

[0158] In some embodiments, the polypeptide comprises a polypeptide having protease activity covalently or non-covalently conjugated to a variant Fc polypeptide, wherein the 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, and further comprises one or more mutations at positions 234, 235, 237, 296, 329, or any combination thereof.

[0159] In some embodiments, the polypeptide comprises a polypeptide having protease activity covalently or non-covalently conjugated to a variant Fc polypeptide, wherein the 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, and further comprises one or more mutations at positions 234, 235, 237, 296, 329, or any combination thereof.

[0160] In some embodiments, the polypeptide comprises a polypeptide having protease activity covalently or non-covalently conjugated to a variant Fc polypeptide, wherein the 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 any of the sequences provided in Table 3, and further comprises one or more mutations at positions L234, L235, G237, Y296, P329, or any combination thereof.

[0161] In some embodiments, the polypeptide comprises a polypeptide having protease activity covalently or non-covalently conjugated to a variant Fc polypeptide, wherein the 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, and further comprises one or more mutations at positions L234, L235, G237, Y296, P329, or any combination thereof.

[0162] In some embodiments, the polypeptide comprises a polypeptide having protease activity covalently or non-covalently conjugated to a variant Fc polypeptide, wherein the 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, and further comprises one or more mutations at positions L234, L235, G237, Y296, P329, or any combination thereof.

[0163] In some embodiments, the polypeptide comprises a polypeptide with 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 any of the sequences provided in Table 4, Table 5, Table 7, or Table 8, wherein the polypeptide with protease activity is covalently linked to a variant Fc polypeptide. Covalently or non-covalently conjugated, the 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 any of the sequences provided in Table 3, and further comprises one or more mutations at 234, 235, 237, 296, 329, or any combination thereof.

[0164] In some embodiments, the polypeptide 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: 26, 28, 29, 31, 36, 37, 38, 39, 40, 41, 50, 51, 52, 55-1019, 1290-1310, or 1333, is covalently or non-covalently conjugated to a variant Fc polypeptide, wherein the 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, and further comprises one or more mutations at 234, 235, 237, 296, 329, or any combination thereof.

[0165] In some embodiments, the polypeptide 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: 26, 28, 29, 31, 36, 37, 38, 39, 40, 41, 50, 51, 52, 55-1019, 1290-1310, or 1333, is covalently or non-covalently conjugated to a variant Fc polypeptide, wherein the 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, and comprises one or more mutations at positions 234, 235, 237, 296, 329, or any combination thereof.

[0166] In some embodiments, the polypeptide comprises a polypeptide with 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 any of the sequences provided in Table 4, Table 5, Table 7, or Table 8, wherein the polypeptide with protease activity is covalently linked to the variant Fc polypeptide. The variant Fc polypeptide is covalently or non-covalently conjugated to any of the sequences provided in Table 3, and 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 of the sequences provided in Table 3, and further comprises one or more mutations at L234, L235, G237, Y296, P329, or any combination thereof.

[0167] In some embodiments, the polypeptide comprises a polypeptide with 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: 26, 28, 29, 31, 36, 37, 38, 39, 40, 41, 50, 51, 52, 55-1019, 1290-1310, or 1333, wherein the polypeptide with protease activity is a variant. The variant Fc polypeptide is covalently or non-covalently conjugated to a variant Fc polypeptide, wherein the 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, and further comprises one or more mutations at L234, L235, G237, Y296, P329, or any combination thereof.

[0168] In some embodiments, the polypeptide 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: 26, 28, 29, 31, 36, 37, 38, 39, 40, 41, 50, 51, 52, 55-1019, 1290-1310, or 1333, The polypeptide is covalently or non-covalently conjugated to a variant Fc polypeptide, wherein the 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 and comprises one or more mutations at positions L234, L235, G237, Y296, P329, or any combination thereof.

[0169] In some embodiments, the polypeptide comprises a polypeptide having protease activity covalently or non-covalently conjugated to a variant Fc polypeptide, wherein the 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 any of the sequences provided in Table 3, and further comprises one or more mutations: L234A, L235A, G237A, Y296Q, P329K, or any combination thereof.

[0170] In some embodiments, the polypeptide comprises a polypeptide having protease activity covalently or non-covalently conjugated to a variant Fc polypeptide, wherein the 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, and further comprises one or more mutations of L234A, L235A, G237A, Y296Q, P329K, or any combination thereof.

[0171] In some embodiments, the polypeptide comprises a polypeptide with 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 any of the sequences provided in Table 4, Table 5, Table 7, or Table 8, wherein the polypeptide with protease activity is covalently linked to the variant Fc polypeptide. or non-covalently conjugated, wherein the 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 any of the sequences provided in Table 3, and further comprises one or more mutations: L234A, L235A, G237A, Y296Q, P329K, or any combination thereof.

[0172] In some embodiments, the polypeptide comprises a polypeptide with 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: 26, 28, 29, 31, 36, 37, 38, 39, 40, 41, 50, 51, 52, 55-1019, 1290-1310, or 1333, wherein the polypeptide with protease activity comprises a mutated The variant Fc polypeptide is covalently or non-covalently conjugated to an Fc polypeptide, wherein the 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, and further comprises one or more mutations: L234A, L235A, G237A, Y296Q, P329K, or any combination thereof.

[0173] In some embodiments, the polypeptide comprises a polypeptide with 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: 26, 28, 29, 31, 36, 37, 38, 39, 40, 41, 50, 51, 52, 55-1019, 1290-1310, or 1333, wherein the polypeptide with protease activity comprises a mutated The variant Fc polypeptide is covalently or non-covalently conjugated to an Fc polypeptide, wherein the 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, and further comprises one or more mutations: L234A, L235A, G237A, Y296Q, P329K, or any combination thereof.

[0174] In some embodiments, the polypeptide 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 NOs: 26, 28, 29, 31, 36, 37, 38, 39, 40, 41, 50, 51, 52, 55-1019, 1290-1310, or 1333, The polypeptide is covalently or non-covalently conjugated to a variant Fc polypeptide, wherein the 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, and further comprises one or more mutations: L234A, L235A, G237A, Y296Q, P329K, or any combination thereof.

[0175] In some embodiments, the polypeptide comprises a polypeptide with protease activity covalently or non-covalently conjugated to a variant Fc polypeptide, wherein the variant Fc polypeptide comprises the amino acid sequence of SEQ ID NO: 19, 20, 21, 22, 23, 24, 25, 42, 43, 44, 45, 46, 47, 48, or 49. In some embodiments, the polypeptide comprises a polypeptide with protease activity covalently or non-covalently conjugated to a variant Fc polypeptide, wherein the variant Fc polypeptide comprises the amino acid sequence of SEQ ID NO: 19. In some embodiments, the polypeptide comprises a polypeptide with protease activity covalently or non-covalently conjugated to a variant Fc polypeptide, wherein the variant Fc polypeptide comprises the amino acid sequence of SEQ ID NO: 20. In some embodiments, the polypeptide comprises a polypeptide with protease activity covalently or non-covalently conjugated to a variant Fc polypeptide, wherein the variant Fc polypeptide comprises the amino acid sequence of SEQ ID NO: 21. In some embodiments, the polypeptide comprises a polypeptide with protease activity covalently or non-covalently conjugated to a variant Fc polypeptide, wherein the variant Fc polypeptide comprises the amino acid sequence of SEQ ID NO: 22. In some embodiments, the polypeptide comprises a polypeptide with protease activity covalently or non-covalently conjugated to a variant Fc polypeptide, wherein the variant Fc polypeptide comprises the amino acid sequence of SEQ ID NO: 23. In some embodiments, the polypeptide comprises a polypeptide with protease activity covalently or non-covalently conjugated to a variant Fc polypeptide, wherein the variant Fc polypeptide comprises the amino acid sequence of SEQ ID NO: 24.In some embodiments, the polypeptide comprises a polypeptide with protease activity covalently or non-covalently conjugated to a variant Fc polypeptide, wherein the variant Fc polypeptide comprises the amino acid sequence of SEQ ID NO: 25. In some embodiments, the polypeptide comprises a polypeptide with protease activity covalently or non-covalently conjugated to a variant Fc polypeptide, wherein the variant Fc polypeptide comprises the amino acid sequence of SEQ ID NO: 42. In some embodiments, the polypeptide comprises a polypeptide with protease activity covalently or non-covalently conjugated to a variant Fc polypeptide, wherein the variant Fc polypeptide comprises the amino acid sequence of SEQ ID NO: 43. In some embodiments, the polypeptide comprises a polypeptide with protease activity covalently or non-covalently conjugated to a variant Fc polypeptide, wherein the variant Fc polypeptide comprises the amino acid sequence of SEQ ID NO: 44. In some embodiments, the polypeptide comprises a polypeptide with protease activity covalently or non-covalently conjugated to a variant Fc polypeptide, wherein the variant Fc polypeptide comprises the amino acid sequence of SEQ ID NO: 45. In some embodiments, the polypeptide comprises a polypeptide with protease activity covalently or non-covalently conjugated to a variant Fc polypeptide, wherein the variant Fc polypeptide comprises the amino acid sequence of SEQ ID NO: 46. In some embodiments, the polypeptide comprises a polypeptide with protease activity covalently or non-covalently conjugated to a variant Fc polypeptide, wherein the variant Fc polypeptide comprises the amino acid sequence of SEQ ID NO: 47. In some embodiments, the polypeptide comprises a polypeptide with protease activity covalently or non-covalently conjugated to a variant Fc polypeptide, wherein the variant Fc polypeptide comprises the amino acid sequence of SEQ ID NO: 48. In some embodiments, the polypeptide comprises a polypeptide with protease activity covalently or non-covalently conjugated to a variant Fc polypeptide, wherein the variant Fc polypeptide comprises the amino acid sequence of SEQ ID NO: 49.

[0176] In some embodiments, the polypeptide comprises a polypeptide with protease activity comprising any of the sequences provided in Table 4, Table 5, Table 7, or Table 8, wherein the polypeptide with protease activity is covalently or non-covalently conjugated to a variant Fc polypeptide, wherein the variant Fc polypeptide comprises an amino acid sequence provided in Table 3. In some embodiments, the polypeptide comprises a polypeptide with protease activity comprising the amino acid sequence of SEQ ID NO: 26, 28, 29, 31, 36, 37, 38, 39, 40, 41, 50, 51, 52, 55-1019, 1290-1310, or 1333, wherein the polypeptide with protease activity is covalently or non-covalently conjugated to a variant Fc polypeptide, wherein the variant Fc polypeptide comprises the amino acid sequence of SEQ ID NO: 19, 20, 21, 22, 23, 24, 25, 42, 43, 44, 45, 46, 47, 48, or 49.

[0177] In some embodiments, the polypeptide comprises a polypeptide with protease activity comprising the amino acid sequence of SEQ ID NO: 26, 28, 29, 31, 36, 37, 38, 39, 40, 41, 50, 51, 52, 55-1019, 1290-1310, or 1333, wherein the polypeptide with protease activity is covalently or non-covalently conjugated to a variant Fc polypeptide, wherein the variant Fc polypeptide comprises the amino acid sequence of SEQ ID NO: 23.

[0178] In some embodiments, the polypeptide comprises a polypeptide with protease activity covalently or non-covalently conjugated to a variant Fc polypeptide, wherein the variant Fc polypeptide comprises an amino acid sequence having a C-terminal lysine (K).

[0179] In some embodiments, the polypeptide comprises a polypeptide with protease activity covalently or non-covalently conjugated to a variant Fc polypeptide, wherein the variant Fc polypeptide comprises an amino acid sequence that does not have a C-terminal lysine (K).

[0180] In some embodiments, the histidine tag comprises the amino acid sequence of SEQ ID NO: 27 or 34. In some embodiments, the histidine tag comprises the amino acid sequence of SEQ ID NO: 27. In some embodiments, the histidine tag comprises the amino acid sequence of SEQ ID NO: 34.

[0181] In some embodiments, the polypeptide has, from N-terminus to C-terminus, the following formula: P1---mutant Fc and During the ceremony, P1 comprises a polypeptide having protease activity, The variant Fc is any variant IgG Fc provided herein.

[0182] In some embodiments, the polypeptide has, from N-terminus to C-terminus, the following formula: Mutant Fc---P1 and During the ceremony, P1 comprises a polypeptide having protease activity, The variant Fc is any variant IgG Fc provided herein.

[0183] In some embodiments, the polypeptide has, from N-terminus to C-terminus, the following formula: P1---mutant Fc and During the ceremony, P1 comprises a polypeptide having protease activity, The variant Fc is any variant IgG Fc provided herein; Optionally, the protease is a glycosylation-resistant protease.

[0184] In some embodiments, the polypeptide has, from N-terminus to C-terminus, the following formula: Mutant Fc---P1 and During the ceremony, P1 comprises a polypeptide having protease activity, The variant Fc is any variant IgG Fc provided herein; Optionally, the protease is a glycosylation-resistant protease.

[0185] In some embodiments, the polypeptide has, from N-terminus to C-terminus, the following formula: N1---P1---mutant Fc and During the ceremony, P1 comprises a polypeptide having protease activity, N1 comprises a nanobody, The variant Fc is any variant IgG Fc provided herein.

[0186] In some embodiments, the polypeptide has, from N-terminus to C-terminus, the following formula: Mutant Fc---P1---N1 and During the ceremony, P1 comprises a polypeptide having protease activity, N1 comprises a nanobody, The variant Fc is any variant IgG Fc provided herein.

[0187] In some embodiments, the polypeptide has, from N-terminus to C-terminus, the following formula: N1---mutant Fc---P1 and During the ceremony, P1 comprises a polypeptide having protease activity, N1 comprises a nanobody, The variant Fc is any variant IgG Fc provided herein.

[0188] In some embodiments, the polypeptide has, from N-terminus to C-terminus, the following formula: P1---mutant Fc---N1 and During the ceremony, P1 comprises a polypeptide having protease activity, N1 comprises a nanobody, The variant Fc is any variant IgG Fc provided herein.

[0189] In some embodiments, the polypeptide has, from N-terminus to C-terminus, the following formula: P1---N1 and During the ceremony, P1 comprises a polypeptide having protease activity, N1 includes nanobodies.

[0190] In some embodiments, the polypeptide has, from N-terminus to C-terminus, the following formula: N1---P1 and During the ceremony, P1 comprises a polypeptide having protease activity, N1 includes nanobodies.

[0191] In some embodiments, the polypeptide has, from N-terminus to C-terminus, the following formula: N1---P1---mutant Fc and During the ceremony, P1 comprises a polypeptide having protease activity, N1 comprises a nanobody, The variant Fc is any variant IgG Fc provided herein; Optionally, the protease is a glycosylation-resistant protease.

[0192] In some embodiments, the polypeptide has, from N-terminus to C-terminus, the following formula: Mutant Fc---P1---N1 and During the ceremony, P1 comprises a polypeptide having protease activity, N1 comprises a nanobody, The variant Fc is any variant IgG Fc provided herein; Optionally, the protease is a glycosylation-resistant protease.

[0193] In some embodiments, the polypeptide has, from N-terminus to C-terminus, the following formula: N1---mutant Fc---P1 and During the ceremony, P1 comprises a polypeptide having protease activity, N1 comprises a nanobody, The variant Fc is any variant IgG Fc provided herein; Optionally, the protease is a glycosylation-resistant protease.

[0194] In some embodiments, the polypeptide has, from N-terminus to C-terminus, the following formula: P1---mutant Fc---N1 and During the ceremony, P1 comprises a polypeptide having protease activity, N1 comprises a nanobody, The variant Fc is any variant IgG Fc provided herein; Optionally, the protease is a glycosylation-resistant protease.

[0195] In some embodiments, the polypeptide has, from N-terminus to C-terminus, the following formula: P1---N1 and During the ceremony, P1 comprises a polypeptide having protease activity, N1 comprises a nanobody, Optionally, the protease is a glycosylation-resistant protease.

[0196] In some embodiments, the polypeptide has, from N-terminus to C-terminus, the following formula: N1---P1 and During the ceremony, P1 comprises a polypeptide having protease activity, N1 comprises a nanobody, Optionally, the protease is a glycosylation-resistant protease.

[0197] In some embodiments, P1 and variant Fc are covalently or non-covalently conjugated to each other according to the formula above. In some embodiments, P1, N1, and variant Fc are covalently or non-covalently conjugated to each other according to the formula above.

[0198] In some embodiments, P1 comprises a polypeptide having protease activity. In some embodiments, N1 comprises a nanobody. In some embodiments, P1 comprises a polypeptide having protease activity and N1 comprises a nanobody.

[0199] In some embodiments, P1 comprises a polypeptide with 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 any sequence provided in Table 4, Table 5, Table 7, or Table 8. In some embodiments, P1 comprises a polypeptide with 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: 26, 28, 29, 31, 36, 37, 38, 39, 40, 41, 50, 51, 52, 55-1019, 1290-1310, or 1333.

[0200] In some embodiments, P1 comprises a polypeptide with protease activity comprising any sequence provided in Table 4, Table 5, Table 7, or Table 8. In some embodiments, P1 comprises a polypeptide with protease activity comprising the amino acid sequence of SEQ ID NO: 26, 28, 29, 31, 36, 37, 38, 39, 40, 41, 50, 51, 52, 55-1019, 1290-1310, or 1333.

[0201] In some embodiments, P1 comprises an IdeS, IdeSsuis, IdeE, IdeZ, IdeZ2, IdeE2, Ide85, or IdeC protease. In some embodiments, P1 comprises an IdeS, IdeSsuis, IdeE, IdeZ, IdeZ2, IdeE2, Ide85, or IdeC protease variant. In some embodiments, P1 comprises an IdeS, IdeSsuis, IdeE, IdeZ, IdeZ2, IdeE2, Ide85, or IdeC protease variant 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: 26, 28, 29, 31, 36, 37, 38, 39, 40, 41, 50, 51, 52, 55-1019, 1290-1310, or 1333.

[0202] In some embodiments, P1 comprises an IdeS, IdeSsuis, IdeE, IdeZ, IdeZ2, IdeE2, Ide85, or IdeC protease variant comprising the amino acid sequence of SEQ ID NO: 26, 28, 29, 31, 36, 37, 38, 39, 40, 41, 50, 51, 52, 55-1019, 1290-1310, or 1333.

[0203] In some embodiments, the variant Fc comprises a variant Fc polypeptide. In some embodiments, the variant Fc polypeptide is such as those provided herein. In some embodiments, the 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, 25, 42, 43, 44, 45, 47, 48, or 49. In some embodiments, the 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.

[0204] In some embodiments, a variant Fc polypeptide comprises the amino acid sequence of SEQ ID NO: 19, 20, 21, 22, 23, 24, 25, 42, 43, 44, 45, 47, 48, or 49. In some embodiments, a variant Fc polypeptide comprises the amino acid sequence of SEQ ID NO: 19. In some embodiments, a variant Fc polypeptide comprises the amino acid sequence of SEQ ID NO: 20. In some embodiments, a variant Fc polypeptide comprises the amino acid sequence of SEQ ID NO: 21. In some embodiments, a variant Fc polypeptide comprises the amino acid sequence of SEQ ID NO: 22. In some embodiments, a variant Fc polypeptide comprises the amino acid sequence of SEQ ID NO: 23. In some embodiments, a variant Fc polypeptide comprises the amino acid sequence of SEQ ID NO: 24. In some embodiments, a variant Fc polypeptide comprises the amino acid sequence of SEQ ID NO: 25. In some embodiments, a variant Fc polypeptide comprises the amino acid sequence of SEQ ID NO: 42. In some embodiments, a variant Fc polypeptide comprises the amino acid sequence of SEQ ID NO: 43. In some embodiments, a variant Fc polypeptide comprises the amino acid sequence of SEQ ID NO: 44. In some embodiments, a variant Fc polypeptide comprises the amino acid sequence of SEQ ID NO: 45. In some embodiments, the variant Fc polypeptide comprises the amino acid sequence of SEQ ID NO: 46. In some embodiments, the variant Fc polypeptide comprises the amino acid sequence of SEQ ID NO: 47. In some embodiments, the variant Fc polypeptide comprises the amino acid sequence of SEQ ID NO: 48. In some embodiments, the variant Fc polypeptide comprises the amino acid sequence of SEQ ID NO: 49.

[0205] In some embodiments, N1 comprises a Nanobody comprising an amino acid sequence that has 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: 30, 32, or 33. In some embodiments, N1 comprises a Nanobody comprising an amino acid sequence that has 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: 30. In some embodiments, N1 comprises a Nanobody comprising an amino acid sequence that has 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: 32. In some embodiments, N1 comprises a Nanobody comprising an amino acid sequence that has 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: 33.

[0206] In some embodiments, N1 comprises a Nanobody comprising the amino acid sequence of SEQ ID NO: 30, 32, or 33. In some embodiments, N1 comprises a Nanobody comprising the amino acid sequence of SEQ ID NO: 30. In some embodiments, N1 comprises a Nanobody comprising the amino acid sequence of SEQ ID NO: 32. In some embodiments, N1 comprises a Nanobody comprising the amino acid sequence of SEQ ID NO: 33.

[0207] In some embodiments, R4 comprises a Nanobody comprising an amino acid sequence that has 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: 30, 32, or 33. In some embodiments, R4 comprises a Nanobody comprising an amino acid sequence that has 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: 30. In some embodiments, R4 comprises a Nanobody comprising an amino acid sequence that has 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: 32. In some embodiments, R4 comprises a Nanobody comprising an amino acid sequence that has 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: 33.

[0208] In some embodiments, R4 comprises a Nanobody comprising the amino acid sequence of SEQ ID NO: 30, 32, or 33. In some embodiments, R4 comprises a Nanobody comprising the amino acid sequence of SEQ ID NO: 30. In some embodiments, R4 comprises a Nanobody comprising the amino acid sequence of SEQ ID NO: 32. In some embodiments, R4 comprises a Nanobody comprising the amino acid sequence of SEQ ID NO: 33.

[0209] In some embodiments, from N-terminus to C-terminus, the formula: N1---P1---Mutant Fc---R2---R4 The polypeptide having the formula (I) further comprises a histidine tag (His tag), wherein the His tag is covalently or non-covalently conjugated to the C-terminus of the polypeptide. In some embodiments, the His tag is such as those provided herein. In some embodiments, the His tag comprises the sequence HHHHHH (SEQ ID NO: 34) or HHHHHHHH (SEQ ID NO: 27).

[0210] In some embodiments, the molecule comprises: a) a first Fc 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% sequence identity to SEQ ID NO:1 (WT), wherein the Fc polypeptide has 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% sequence identity to SEQ ID NO:1 (WT), and a first Fc polypeptide comprising a mutation or set of mutations at any one position or any combination of positions selected from 234, 235, 237, 296, and 329, ... mutation or set of mutations at any one position or any combination of positions selected from 234, 235, 237, 296, and 329, and a mutation or set of mutation %, or at least 99% sequence identity to SEQ ID NO: 35, wherein the Fc polypeptide comprises a mutation or set of mutations at any one position or any combination of positions selected from 234, 235, 237, 296, and 329 relative to SEQ ID NO: 1, and wherein the first Fc polypeptide and the second Fc polypeptide associate with each other to form a dimeric Fc molecule; and b) a second Fc polypeptide comprising an amino acid sequence having at least 50%, at least 55%, at least 60%, or at least 99% sequence identity to SEQ ID NO: 35, wherein the Fc polypeptide comprises a mutation or set of mutations at any one position or any combination of positions selected from 234, 235, 237, 296, and 329 relative to SEQ ID NO: 1, and wherein the first Fc polypeptide and the second Fc polypeptide associate with each other to form a dimeric Fc molecule. 10. A polypeptide comprising an amino acid sequence having 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% sequence identity, with respect to SEQ ID NO: 35, wherein the polypeptide is 60, 61, 63, 64, 69, 74, 76, 85, 112, 115, 116, 119, 120, 121, 130, 134, 145, 146, 147, 148, 149, 150, 151, 152, 153, 154, 155, 156, 157, 158, 159, 160, 161, 162, 163, 164, 165, 166, 167, 168, 169, 200, 201, 202, 203, 204, 205, 206, 207, 208, 209, 210, 211, 212, 213, 214, 215, 216, 217, 218, 219, 220, 221, 222, 223, 224, 225, 226, 227, 228, 229, 230, 231, 232, 233, 234, 235, 236, 237, 2a polypeptide comprising a mutation or set of mutations at any one position, or any combination of positions, selected from: 146, 154, 190, 198, 223, 225, 227, 231, 241, 251, 258, 261, 273, 274, 278, 286, 289, 303, 314, 334, and 336, wherein the polypeptide is linked or conjugated to the N-terminus or C-terminus of the first Fc polypeptide and / or the second Fc polypeptide;

[0211] In some embodiments, the dimer is a homodimer. In some embodiments, the dimer is a heterodimer. In some embodiments, the first Fc polypeptide comprises a set of mutations at positions 234, 235, 237, 296, and 329 relative to SEQ ID NO:1.

[0212] In some embodiments, the second Fc polypeptide comprises a set of mutations at positions 234, 235, 237, 296, and 329 compared to SEQ ID NO:1.

[0213] In some embodiments, the first Fc polypeptide comprises a set of mutations at positions 234, 235, 237, 296, and 329 compared to SEQ ID NO:1, and the second Fc polypeptide comprises a set of mutations at positions 234, 235, 237, 296, and 329 compared to SEQ ID NO:1.

[0214] In some embodiments, the molecule comprises: a) a first Fc 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% sequence identity to SEQ ID NO:23, with the proviso that the Fc polypeptide has ... and a first Fc polypeptide comprising a mutation or set of mutations at any one position or any combination of positions selected from 234, 235, 237, 296, and 329, and a first Fc polypeptide comprising a mutation or set of mutations at any one position or any combination of positions selected from 234, 235, 237, 296, and 329 that is 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 and b) a second Fc polypeptide comprising an amino acid sequence having at least 99% sequence identity to SEQ ID NO: 1, wherein the Fc polypeptide comprises a mutation or set of mutations at any one position or any combination of positions selected from 234, 235, 237, 296, and 329 relative to SEQ ID NO: 1, and wherein the first Fc polypeptide and the second Fc polypeptide associate with each other to form a dimeric Fc molecule; and 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 compared to SEQ ID NO: 35, provided that the polypeptide is 60, 61, 63, 64, 69, 74, 76, 85, 112, 115, 116, 119, 200, 201, 202, 203, 204, 205, 206, 207, 208, 209, 210, 211, 212, 213, 214, 215, 216, 217, 218, 219, 220, 221, 222, 223, 224, 225, 226, 227, 228, 229, 230, 231, 232, 233, 234, 235, 236, 237, 238, 239, 240, 241, 242, 243, 244, 245, 246, 247, 248, 249, 250, 251, 252, 253, 254, 255, 256, 257, 258, 259, 260, 261, 262, 263, 264, 265, 266, 267, 268, 269, 300, 301, 302, 303, 304, 305, 3a polypeptide comprising a mutation or set of mutations at any one position, or any combination of positions, selected from 120, 121, 130, 134, 145, 146, 154, 190, 198, 223, 225, 227, 231, 241, 251, 258, 261, 273, 274, 278, 286, 289, 303, 314, 334, and 336, wherein the polypeptide is linked or conjugated to the N-terminus or C-terminus of the first Fc polypeptide and / or the second Fc polypeptide;

[0215] In some embodiments, the molecule comprises a dimeric Fc molecule comprising a first Fc polypeptide having the amino acid sequence of SEQ ID NO: 23 and a second Fc polypeptide having the 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 dimeric Fc molecule; and a polypeptide having the amino acid sequence of any one of SEQ ID NOs: 1290, 1295, 1301, 1303, and 1310, or any combination thereof, wherein the polypeptide is linked or conjugated to the N-terminus or C-terminus of the first Fc polypeptide and / or the second Fc polypeptide.

[0216] In some embodiments, the molecule comprises a dimeric Fc molecule comprising a first Fc polypeptide having the amino acid sequence of SEQ ID NO: 23, a second Fc polypeptide having the amino acid sequence of SEQ ID NO: 23, and a polypeptide having the amino acid sequence of SEQ ID NO: 1290, wherein the polypeptides are linked or conjugated to the N-terminus or C-terminus of the first Fc polypeptide and / or the second Fc polypeptide. In some e...

Claims

1. 1. A polypeptide comprising an amino acid sequence at least 50% identical to a polypeptide comprising the amino acid sequence of SEQ ID NO:36, provided that said 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 tyrosine (Y) at position 146, a tyrosine (Y) at position 147, a tyrosine (Y) at position 148, a tyrosine (Y) at position 149, a tyrosine (Y) at position 150, a tyrosine (Y) at position 151, a tyrosine (Y) at position 152, a tyrosine (Y) at position 153, a tyrosine (Y) at position 154, a tyrosine (Y) at position 155, a tyrosine (Y) at position 156, a tyrosine (Y) at position 157, a tyrosine (Y) at position 158, a tyrosine (Y) at position 159, a tyrosine (Y) at position 160, a tyrosine (Y) at position 161, a 35. A polypeptide comprising: serine (S) at position 146, aspartic acid (D) at position 154, aspartic acid (D) at position 190, arginine (R) at position 198, asparagine (N) at position 223, tyrosine (Y) at position 225, leucine (L) at position 227, threonine (T) at position 231, glutamine (Q) at position 241, isoleucine (I) at position 251, glycine (G) at position 261, alanine (A) at position 273, glutamic acid (E) at position 274, glutamic acid (E) at position 278, proline (P) at position 289, serine (S) at position 303, glutamic acid (E) at position 314, and isoleucine (I) at position 334, wherein said positions correspond to those in SEQ ID NO:

35.

2. The polypeptide of claim 1, wherein the polypeptide does not comprise the sequence ANQEIRYSEVTPYHVT (SEQ ID NO: 1353) or ANQEIRYSEVTPYH (SEQ ID NO: 1354).

3. The polypeptide of claim 1, wherein the polypeptide comprises an amino acid sequence that is at least 50% identical to a polypeptide comprising the amino acid sequence of SEQ ID NO: 1349.

4. 4. The polypeptide of claim 3, wherein the polypeptide does not comprise the amino acid sequence of DSFSANQEIRYSEVTPYHVT (SEQ ID NO: 1351) or DSFSANQEIRYSEVTPYH (SEQ ID NO: 1352).

5. The polypeptide comprises an amino acid sequence having at least 80% sequence identity to SEQ ID NO:1303, with the proviso that the polypeptide contains 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, and an asparagine (N) at position 145, as compared to SEQ ID NO:

35.

2. The polypeptide of claim 1, comprising a nucleotide sequence selected from the group consisting of ...

6. 6. The polypeptide of claim 5, wherein the polypeptide comprises a cysteine ​​(C) at position 94 and / or has lysine (K), histidine (H), aspartic acid (D) and / or aspartic acid (D) at positions in the variant sequence corresponding to positions 84, 262, 284 and 286 of SEQ ID NO:

35.

7. The polypeptide is a positively charged amino acid at position 130, optionally arginine (R) or lysine (K), and / or 7. The polypeptide of claim 6, further comprising a positively charged amino acid at position 131, optionally arginine (R) or lysine (K).

8. The polypeptide of claim 1 , wherein the polypeptide has protease activity.

9. The polypeptide of claim 8 , wherein the polypeptide has IgG-cleaving protease activity.

10. The polypeptide of claim 1 , wherein the polypeptide is less immunogenic than wild-type IdeS.

11. The polypeptide of claim 1, wherein the polypeptide comprises the amino acid sequence of SEQ ID NO: 1303.

12. The polypeptide of claim 11, wherein the polypeptide is covalently or non-covalently conjugated to an Fc polypeptide.

13. The polypeptide of claim 12, wherein the Fc polypeptide comprises an amino acid sequence having at least 90% sequence identity to SEQ ID NO:

23.

14. The polypeptide of claim 13, wherein the Fc polypeptide comprises the amino acid sequence of SEQ ID NO:

23.

15. 13. The polypeptide of claim 12, wherein the polypeptide is covalently or non-covalently conjugated to the N-terminus or C-terminus of the Fc polypeptide via a linker.

16. The polypeptide of claim 15 , wherein the linker is a peptide linker.

17. The polypeptide of claim 12, wherein the polypeptide comprises the amino acid sequence of SEQ ID NO: 1318.

18. The polypeptide of claim 12, wherein the Fc polypeptide self-associates to form a dimer.

19. 1303, 1290, 1291, 1292, 1293, 1294, 1295, 1296, 1297, 1298, 1299, 1300, 1301, 1302, 1304, 1305, 1306, 1307, 1308, 1309, 1310, and 1333, provided that the polypeptide has at least 50% sequence identity between position 60 and 61 compared to SEQ ID NO:

35. threonine (T) at position 61, aspartic acid (D) at position 61, isoleucine (I) at position 63, tyrosine (Y) at position 64, leucine (L) at position 69, lysine (K) at position 74, glycine (G) at position 76, leucine (L) at position 85, glutamic acid (E) at position 112, glutamic acid (E) at position 115, asparagine (N) at position 116, lysine (K) at position 119, lysine (K) at position 120, tyrosine (Y) at position 121, Lysine (K) at position 130, glycine (G) at position 130, leucine (L) at position 134, aspartic acid (D) at position 145, serine (S) at position 146, aspartic acid (D) at position 154, aspartic acid (D) at position 190, arginine (R) at position 198, asparagine (N) at position 223, tyrosine (Y) at position 225, leucine (L) at position 227, threonine (T) at position 231, glutamine (Q) at position 241, A polypeptide comprising any one or any combination of isoleucine (I) at position 251, glycine (G) at position 261, alanine (A) at position 273, glutamic acid (E) at position 274, glutamic acid (E) at position 278, asparagine (N) at position 286, proline (P) at position 289, serine (S) at position 303, glutamic acid (E) at position 314, isoleucine (I) at position 334, and glutamine (Q) at position 336.

20. The polypeptide is Compared to SEQ ID NO:35, there is 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, a glutamic acid (E) at position 115, 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 lysine (K) at position 130, a leucine (L) at position 134, an aspartic acid (D) at position 145, a serine ( S), aspartic acid (D) at position 154, aspartic acid (D) at position 190, arginine (R) at position 198, asparagine (N) at position 223, tyrosine (Y) at position 225, leucine (L) at position 227, threonine (T) at position 231, glutamine (Q) at position 241, isoleucine (I) at position 251, alanine (A) at position 273, glutamic acid (E) at position 274, glutamic acid (E) at position 278, proline (P) at position 289, serine (S) at position 303, glutamic acid (E) at position 314, isoleucine (I) at position 334, and glutamine (Q) at position 336, Compared to SEQ ID NO: 35, threonine (T) at position 60, aspartic acid (D) at position 61, isoleucine (I) at position 63, tyrosine (Y) at position 64, leucine (L) at position 69, lysine (K) at position 74, glycine (G) at position 76, leucine (L) at position 85, glutamic acid (E) at position 112, glutamic acid (E) at position 115, asparagine (N) at position 116, lysine (K) at position 119, lysine (K) at position 120, tyrosine (Y) at position 121, lysine (K) at position 130, leucine (L) at position 134, aspartic acid (D) at position 145, serine (S) at position 146, aspartic acid (D) at position 4, aspartic acid (D) at position 190, arginine (R) at position 198, asparagine (N) at position 223, tyrosine (Y) at position 225, leucine (L) at position 227, threonine (T) at position 231, glutamine (Q) at position 241, isoleucine (I) at position 251, glycine (G) at position 258, alanine (A) at position 273, glutamic acid (E) at position 274, glutamic acid (E) at position 278, proline (P) at position 289, serine (S) at position 303, glutamic acid (E) at position 314, isoleucine (I) at position 334, and glutamine (Q) at position 336, Compared to SEQ ID NO: 35, threonine (T) at position 60, aspartic acid (D) at position 61, isoleucine (I) at position 63, tyrosine (Y) at position 64, leucine (L) at position 69, lysine (K) at position 74, glycine (G) at position 76, leucine (L) at position 85, glutamic acid (E) at position 112, glutamic acid (E) at position 115, asparagine (N) at position 116, lysine (K) at position 119, lysine (K) at position 120, tyrosine (Y) at position 121, leucine (L) at position 134, aspartic acid (D) at position 145, serine (S) at position 146, asparagine at position 154 acid (D) at position 190, aspartic acid (D) at position 190, arginine (R) at position 198, asparagine (N) at position 223, tyrosine (Y) at position 225, leucine (L) at position 227, threonine (T) at position 231, glutamine (Q) at position 241, isoleucine (I) at position 251, alanine (A) at position 273, glutamic acid (E) at position 274, glutamic acid (E) at position 278, asparagine (N) at position 286, proline (P) at position 289, serine (S) at position 303, glutamic acid (E) at position 314, isoleucine (I) at position 334, and glutamine (Q) at position 336, Compared to SEQ ID NO: 35, threonine (T) at position 60, aspartic acid (D) at position 61, isoleucine (I) at position 63, tyrosine (Y) at position 64, leucine (L) at position 69, lysine (K) at position 74, glycine (G) at position 76, leucine (L) at position 85, glutamic acid (E) at position 112, asparagine (N) at position 116, lysine (K) at position 119, lysine (K) at position 120, tyrosine (Y) at position 121, glycine (G) at position 130, leucine (L) at position 134, aspartic acid (D) at position 145, serine (S) at position 146, and leucine (L) at position 154. aspartic acid (D) at position 190, aspartic acid (D) at position 190, arginine (R) at position 198, asparagine (N) at position 223, tyrosine (Y) at position 225, leucine (L) at position 227, threonine (T) at position 231, glutamine (Q) at position 241, isoleucine (I) at position 251, glycine (G) at position 261, alanine (A) at position 273, glutamic acid (E) at position 274, glutamic acid (E) at position 278, proline (P) at position 289, serine (S) at position 303, glutamic acid (E) at position 314, and isoleucine (I) at position 334, or Compared to SEQ ID NO:35, there is 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, a glutamic acid (E) at position 115, 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 lysine (K) at position 130, a leucine (L) at position 134, an aspartic acid (D) at position 145, a serine (S) at position 146, an aspartic acid ( 20. The polypeptide of claim 19, comprising: an amino acid sequence selected from the group consisting of ...

21. 20. The polypeptide of claim 19, wherein the polypeptide comprises a cysteine ​​(C) at position 94 and / or has lysine (K), histidine (H), aspartic acid (D) and / or aspartic acid (D) at positions in the variant sequence corresponding to positions 84, 262, 284 and 286 of SEQ ID NO: 35, respectively.

22. The polypeptide is a positively charged amino acid at position 130 (optionally, said positively charged amino acid is arginine (R) or lysine (K)), and / or 21. The polypeptide of claim 20, further comprising a positively charged amino acid at position 131 (optionally, the positively charged amino acid is arginine (R) or lysine (K)).

23. 20. The polypeptide of claim 19, wherein the polypeptide has protease activity.

24. 24. The polypeptide of claim 23, wherein the polypeptide has IgG-cleaving protease activity.

25. 20. The polypeptide of claim 19, wherein the polypeptide is less immunogenic than wild-type IdeS.

26. 24. The polypeptide of any one of claims 19 to 23, wherein the polypeptide comprises the amino acid sequence of any one of SEQ ID NOs: 1303, 1290, 1291, 1292, 1293, 1294, 1295, 1296, 1297, 1298, 1299, 1300, 1301, 1302, 1304, 1305, 1306, 1307, 1308, 1309, 1310, and 1333.

27. 20. The polypeptide of claim 19, wherein the polypeptide is covalently or non-covalently conjugated to an Fc polypeptide.

28. 28. The polypeptide of claim 27, wherein the Fc polypeptide comprises an amino acid sequence having at least 90% sequence identity to any one of SEQ ID NOs: 19, 20, 21, 22, 23, 24, or 25.

29. 29. The polypeptide of claim 28, wherein the Fc polypeptide comprises the amino acid sequence of any one of SEQ ID NOs: 19, 20, 21, 22, 23, 24, or 25.

30. 28. The polypeptide of claim 27, wherein the polypeptide is covalently or non-covalently conjugated to the N-terminus or C-terminus of the Fc polypeptide via a linker.

31. 31. The polypeptide of claim 30, wherein the linker is a peptide linker.

32. The polypeptide comprises, from the N-terminus to the C-terminus: the Fc polypeptide having the amino acid sequence of SEQ ID NO: 23, a peptide linker, and the polypeptide having the amino acid sequence of SEQ ID NO: 1303; the Fc polypeptide having the amino acid sequence of SEQ ID NO:23, a peptide linker, and the polypeptide having the amino acid sequence of SEQ ID NO:1290; the Fc polypeptide having the amino acid sequence of SEQ ID NO: 23, a peptide linker, and the polypeptide having the amino acid sequence of SEQ ID NO: 1291; the Fc polypeptide having the amino acid sequence of SEQ ID NO:23, a peptide linker, and the polypeptide having the amino acid sequence of SEQ ID NO:1292; the Fc polypeptide having the amino acid sequence of SEQ ID NO: 23, a peptide linker, and the polypeptide having the amino acid sequence of SEQ ID NO: 1293; the Fc polypeptide having the amino acid sequence of SEQ ID NO:23, a peptide linker, and the polypeptide having the amino acid sequence of SEQ ID NO:1294; the Fc polypeptide having the amino acid sequence of SEQ ID NO:23, a peptide linker, and the polypeptide having the amino acid sequence of SEQ ID NO:1295; the Fc polypeptide having the amino acid sequence of SEQ ID NO:23, a peptide linker, and the polypeptide having the amino acid sequence of SEQ ID NO:1296; the Fc polypeptide having the amino acid sequence of SEQ ID NO:23, a peptide linker, and the polypeptide having the amino acid sequence of SEQ ID NO:1297; the Fc polypeptide having the amino acid sequence of SEQ ID NO:23, a peptide linker, and the polypeptide having the amino acid sequence of SEQ ID NO:1298; the Fc polypeptide having the amino acid sequence of SEQ ID NO:23, a peptide linker, and the polypeptide having the amino acid sequence of SEQ ID NO:1299; the Fc polypeptide having the amino acid sequence of SEQ ID NO: 23, a peptide linker, and the polypeptide having the amino acid sequence of SEQ ID NO: 1300; the Fc polypeptide having the amino acid sequence of SEQ ID NO: 23, a peptide linker, and the polypeptide having the amino acid sequence of SEQ ID NO: 1301; the Fc polypeptide having the amino acid sequence of SEQ ID NO: 23, a peptide linker, and the polypeptide having the amino acid sequence of SEQ ID NO: 1302; the Fc polypeptide having the amino acid sequence of SEQ ID NO: 23, a peptide linker, and the polypeptide having the amino acid sequence of SEQ ID NO: 1304; the Fc polypeptide having the amino acid sequence of SEQ ID NO: 23, a peptide linker, and the polypeptide having the amino acid sequence of SEQ ID NO: 1305; the Fc polypeptide having the amino acid sequence of SEQ ID NO: 23, a peptide linker, and the polypeptide having the amino acid sequence of SEQ ID NO: 1306; the Fc polypeptide having the amino acid sequence of SEQ ID NO: 23, a peptide linker, and the polypeptide having the amino acid sequence of SEQ ID NO: 1307; the Fc polypeptide having the amino acid sequence of SEQ ID NO: 23, a peptide linker, and the polypeptide having the amino acid sequence of SEQ ID NO: 1308; the Fc polypeptide having the amino acid sequence of SEQ ID NO: 23, a peptide linker, and the polypeptide having the amino acid sequence of SEQ ID NO: 1309; the Fc polypeptide having the amino acid sequence of SEQ ID NO: 23, a peptide linker, and the polypeptide having the amino acid sequence of SEQ ID NO: 1310; the Fc polypeptide having the amino acid sequence of SEQ ID NO: 23, a peptide linker, and the polypeptide having the amino acid sequence of SEQ ID NO: 1333; the polypeptide having the amino acid sequence of SEQ ID NO: 1290, a peptide linker, and the Fc polypeptide having the amino acid sequence of SEQ ID NO: 23; the polypeptide having the amino acid sequence of SEQ ID NO: 1291, a peptide linker, and the Fc polypeptide having the amino acid sequence of SEQ ID NO: 23; the polypeptide having the amino acid sequence of SEQ ID NO: 1292, a peptide linker, and the Fc polypeptide having the amino acid sequence of SEQ ID NO: 23; the polypeptide having the amino acid sequence of SEQ ID NO: 1293, a peptide linker, and the Fc polypeptide having the amino acid sequence of SEQ ID NO: 23; the polypeptide having the amino acid sequence of SEQ ID NO: 1294, a peptide linker, and the Fc polypeptide having the amino acid sequence of SEQ ID NO: 23; the polypeptide having the amino acid sequence of SEQ ID NO: 1295, a peptide linker, and the Fc polypeptide having the amino acid sequence of SEQ ID NO: 23; the polypeptide having the amino acid sequence of SEQ ID NO: 1296, a peptide linker, and the Fc polypeptide having the amino acid sequence of SEQ ID NO: 23; the polypeptide having the amino acid sequence of SEQ ID NO: 1297, a peptide linker, and the Fc polypeptide having the amino acid sequence of SEQ ID NO: 23; the polypeptide having the amino acid sequence of SEQ ID NO: 1298, a peptide linker, and the Fc polypeptide having the amino acid sequence of SEQ ID NO: 23; the polypeptide having the amino acid sequence of SEQ ID NO: 1299, a peptide linker, and the Fc polypeptide having the amino acid sequence of SEQ ID NO: 23; the polypeptide having the amino acid sequence of SEQ ID NO: 1300, a peptide linker, and the Fc polypeptide having the amino acid sequence of SEQ ID NO: 23; the polypeptide having the amino acid sequence of SEQ ID NO: 1301, a peptide linker, and the Fc polypeptide having the amino acid sequence of SEQ ID NO: 23; the polypeptide having the amino acid sequence of SEQ ID NO: 1302, a peptide linker, and the Fc polypeptide having the amino acid sequence of SEQ ID NO: 23; the polypeptide having the amino acid sequence of SEQ ID NO: 1303, a peptide linker, and the Fc polypeptide having the amino acid sequence of SEQ ID NO: 23; the polypeptide having the amino acid sequence of SEQ ID NO: 1304, a peptide linker, and the Fc polypeptide having the amino acid sequence of SEQ ID NO: 23; the polypeptide having the amino acid sequence of SEQ ID NO: 1305, a peptide linker, and the Fc polypeptide having the amino acid sequence of SEQ ID NO: 23; the polypeptide having the amino acid sequence of SEQ ID NO: 1306, a peptide linker, and the Fc polypeptide having the amino acid sequence of SEQ ID NO: 23; the polypeptide having the amino acid sequence of SEQ ID NO: 1307, a peptide linker, and the Fc polypeptide having the amino acid sequence of SEQ ID NO: 23; the polypeptide having the amino acid sequence of SEQ ID NO: 1308, a peptide linker, and the Fc polypeptide having the amino acid sequence of SEQ ID NO: 23; the polypeptide having the amino acid sequence of SEQ ID NO: 1309, a peptide linker, and the Fc polypeptide having the amino acid sequence of SEQ ID NO: 23; the polypeptide having the amino acid sequence of SEQ ID NO: 1310, a peptide linker, and the Fc polypeptide having the amino acid sequence of SEQ ID NO: 23; or 28. The polypeptide of claim 27, comprising the polypeptide having the amino acid sequence of SEQ ID NO: 1333, a peptide linker, and the Fc polypeptide having the amino acid sequence of SEQ ID NO:

23.

33. 28. The polypeptide of claim 27, wherein the Fc polypeptide self-associates to form a dimer.

34. 34. The polypeptide of claim 33, wherein the dimer is a homodimer or a heterodimer.

35. 28. The polypeptide of claim 27, wherein the polypeptide comprises the amino acid sequence of any one of SEQ ID NOs: 1318, 1311, 1312, 1313, 1314, 1315, 1316, 1317, 1319, 1320, 1321, 1322, 1323, 1324, 1325, 1326, 1327, 1328, 1329, 1334, 1335, and 1336.

36. A polypeptide comprising 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 of SEQ ID NOs: 55-1019.

37. The polypeptide of claim 36, wherein the polypeptide has protease activity.

38. 38. The polypeptide of claim 37, wherein the polypeptide has IgG-cleaving protease activity.

39. The polypeptide of any one of claims 36 to 38, wherein the polypeptide comprises the amino acid sequence of any one of SEQ ID NOs: 55 to 1019.

40. 37. The polypeptide of claim 36, wherein the polypeptide is covalently or non-covalently conjugated to an Fc polypeptide.

41. 41. The polypeptide of claim 40, wherein the Fc polypeptide comprises an amino acid sequence having 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 of SEQ ID NOs: 19, 20, 21, 22, 23, 24, or 25.

42. 42. The polypeptide of claim 41, wherein the Fc polypeptide comprises the amino acid sequence of any one of SEQ ID NOs: 19, 20, 21, 22, 23, 24, or 25.

43. 41. The polypeptide of claim 40, wherein the polypeptide is covalently or non-covalently conjugated to the N-terminus or the C-terminus of the Fc polypeptide via a linker.

44. 44. The polypeptide of claim 43, wherein the linker is a peptide linker.

45. 41. The polypeptide of claim 40, wherein the Fc polypeptide self-associates or associates with another Fc polypeptide to form a dimer.

46. 46. ​​The polypeptide of claim 45, wherein the dimer is a homodimer or a heterodimer.

47. The polypeptide of claim 40, wherein the polypeptide comprises the amino acid sequence of any one of SEQ ID NOs: 1023 to 1289.

48. A pharmaceutical composition comprising a polypeptide according to any one of claims 1 to 25 and 27 to 35.

49. A pharmaceutical composition comprising a polypeptide according to any one of claims 1 to 47.

50. A method for reducing immunoglobulins in a subject, comprising administering to the subject a polypeptide described in any one of claims 1 to 25 or 27 to 35, or a pharmaceutical composition comprising the same, thereby reducing immunoglobulins in the subject.

51. 51. The method of claim 50, wherein the immunoglobulin is an IgG immunoglobulin.

52. A method for reducing immunoglobulins in a subject, comprising administering to the subject a polypeptide described in any one of claims 1 to 47, or a pharmaceutical composition comprising the same, thereby reducing immunoglobulins in the subject.

53. A method for treating a disease or disorder in a subject, comprising administering to the subject a polypeptide described in any one of claims 1 to 47, or a pharmaceutical composition comprising the same, thereby treating the disease or disorder.

54. The diseases or disorders include Addison's disease, anti-GBM glomerulonephritis, antineutrophil cytoplasmic antibody-associated vasculitis, ANCA-associated vasculitis, granulomatosis with polyangiitis (Wegener's granulomatosis), eosinophilic granulomatosis with polyangiitis (Churg-Strauss syndrome), microscopic polyangiitis, anti-NMDAR encephalitis, antiphospholipid syndrome (APS) and fulminant APS, autoimmune bullous skin diseases, pemphigus, pemphigus foliaceus (PF), pemphigus braziliensis (FS), pemphigus vulgaris (PV), autoimmune hemolytic anemia (AIHA), autoimmune hepatitis (AIH), autoimmune neutropenia (AIN), bullous granulomatosis with polyangiitis (Wegener's granulomatosis), antiphospholipid syndrome (APS), and fulminant APS. Pemphigus (BP), celiac disease, chronic urticaria, congenital complete atrioventricular block (CCHB), type 1A diabetes mellitus (T1DM), epidermolysis bullosa acquisita (EBA), essential mixed cryoglobulinemia, Goodpasture's syndrome, anti-glomerular basement membrane disease, Graves' disease, goiter, hyperthyroidism, infiltrative exophthalmos, infiltrative dermatosis, Guillain-Barré syndrome (GBS), acute inflammatory demyelinating polyneuropathy (AIDP), acute motor axonal neuropathy (AMAN), hemophilia, acquired FVII deficiency, idiopathic thrombocytopenic purpura (ITP), Lambert-Eaton myasthenia gravis Leukemia-associated 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), Sjögren's syndrome (SS), lupus nephritis, systemic lupus erythematosus (SLE), cutaneous lupus erythematosus (CLE), transplant rejection, thrombotic Thrombocytopenic purpura (TTP), idiopathic inflammatory myopathy, 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 rheumatica, Takayasu's arteritis, antineutrophil cytoplasmic antibody-associated vasculitis, Henoch-Schonlein purpura, polyarteritis nodosa, Cogan's syndrome, Buerger's disease, Susan's disease, immune complex vasculitis, primary central nervous system vasculitis (CNS), Behçet's disease, relapsing polychondritis, juvenile idiopathic arthritis,Juvenile dermatomyositis, autoimmune encephalopathy, sarcoidosis, neurosarcoidosis, IgG4-related disease, immune-related adverse events (IRAEs) of immune checkpoint inhibitors, adult-onset Still's disease, warm antibody hemolytic anemia (wAIHA), immune thrombocytopenia, immune thrombocytopenia, thrombotic thrombocytopenia, pernicious anemia, aplastic anemia, Evans syndrome, autoimmune neutropenia, acquired von Willebrand syndrome , recurrent fetal loss, Rh incompatibility, 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 1 diabetes, Hashimoto's thyroiditis, polyglandular autoimmune endocrine syndrome, atopic dermatitis, dermatitis herpetiformis, pemphigus foliaceus, pemphigus brazilianus, cutaneous lupus erythematosus, linear IgA disease , lichen planus, transplantation, antibody-mediated rejection, alloantibody hypersensitivity, xenoantibody-mediated rejection, solid organ rejection, graft-versus-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, sensitization 54. The method of claim 53, wherein the disease is selected from infection, vaccination, antibody-dependent enhancement, antibodies to therapeutic biologics (cytokines, monoclonal antibodies, enzymes, clotting factors), allergic asthma, eosinophilic pneumonia, nonspecific interstitial pneumonia, rheumatoid arthritis-associated interstitial lung disease (RA-ILD), sarcoidosis, hypersensitivity pneumonitis, allergic bronchopulmonary mycosis, chronic sinusitis with nasal polyps, or any combination thereof.

55. A method for treating a transplant subject, comprising administering to the subject a therapeutically effective amount of a polypeptide described in any one of claims 1 to 47, or a pharmaceutical composition comprising the same, thereby treating the transplant (recipient) subject.

56. A method for improving gene therapy in a subject, comprising administering to the subject a therapeutically effective amount of a polypeptide described in any one of claims 1 to 47, or a pharmaceutical composition comprising same, thereby improving gene therapy in the subject.

57. 50. A method of treating a subject having, or at risk of having, an IgG-mediated disease or disorder, comprising administering a therapeutically effective amount of a polypeptide according to any one of claims 1 to 47, or a pharmaceutical composition comprising same, thereby treating the subject.

58. A method for cleaving a B cell receptor in a subject, the method comprising administering to the subject a therapeutically effective amount of a polypeptide described in any one of claims 1 to 47, or a pharmaceutical composition comprising the same, thereby cleaving the B cell receptor in the subject.

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