Engineered antibodies and uses thereof

Abstract

Disclosed herein are isolated polypeptides and antibodies with improved sensitivity and decreased background signals and methods of using the same. Specifically, the antibodies comprise polypeptides comprising a sequence-defined CH2 domain linked to hinge domain.

Description

Attorney Docket No. TP385350WO1 ENGINEERED ANTIBODIES AND USES THEREOF CROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims the benefit under 35 U.S.C. § 119(e) of Indian Provisional Application no. IN202441066590, filed September 3, 2024, Indian Provisional Application no. IN202341061917, filed September 14, 2023, and Indian Provisional Application no. IN202341061835, filed September 14, 2023. The entire contents of the aforementioned applications are incorporated by reference herein. REFERENCE TO ELECTRONIC SEQUENCE LISTING

[0002] This application contains a Sequence Listing which has been submitted electronically herewith in .XML format and is hereby incorporated by reference in its entirety. Said .XML copy, created on September 10, 2024, is named “TP385350WO1.xml” and is 279,164 bytes in size. The sequence listing contained in this .XML file is part of the specification and is hereby incorporated by reference herein in its entirety. TECHNICAL FIELD

[0003] This invention relates to isolated polypeptides and antibodies with improved sensitivity and methods of use thereof. BACKGROUND

[0004] Recombinant antibodies offer the ability to rationally engineer beneficial properties. One such beneficial property is antibody sensitivity, i.e., the strength of the signal produced in a particular immunoassay. A highly sensitive antibody has several advantages such as the detection of low abundance or rare targets and the ability to detect the target protein in difficult imaging assays. Antibody sensitivity can be increased by affinity maturation of the antibody clone, which involves mutation of the antigen binding site or paratope. However, affinity maturation has to be tailored for each antibody under consideration and is therefore not a scalable solution to address a large number of antibodies. In addition, introduction of mutations to the paratope changes the clone of the antibody andAttorney Docket No. TP385350WO1 essentially produces a new antibody. There is a need for improved approaches for increasing antibody sensitivity by, for example, rational modification of the antibody Fc region. The disclosed peptides, antibodies, compositions and uses thereof are directed to these and other important needs. SUMMARY

[0005] Disclosed herein are isolated polypeptides comprising the amino acid sequence of Formula I: Z-(X-Yn-Zr)p-X, wherein Z is a hinge region, X is a CH2 domain, Y is a linker, n is an integer of 1 or greater, p is an integer of 2 or greater, and r is an integer of 0 or 1. In certain embodiments, the isolated polypeptides have at least a 1.5-fold increase in signal enhancement or sensitivity. In certain embodiments, the isolated polypeptides have at least a 1.5-fold reduction in background signal.

[0006] Disclosed herein are antibodies comprising a polypeptide comprising the amino acid sequence of Formula I: Z-(X-Yn-Zr)p-X, wherein Z is a hinge region, X is a CH2 domain, Y is a linker, n is an integer of 1 or greater, p is an integer of 2 or greater, and r is an integer of 0 or 1 linked to a Fab domain. In certain embodiments, the antibodies have at least a 1.5-fold increase in signal enhancement or sensitivity compared to a corresponding wild type antibody. In certain embodiments, the antibodies have at least a 1.5-fold reduction in background signal compared to a corresponding wild type antibody.

[0007] Disclosed herein are antibodies that immunospecifically bind to Parkin, wherein the antibodies comprise a heavy chain (HC) and a light chain (LC) that pair to form a binding domain comprising a heavy chain complementarity determining region 1 (HCDR1) comprising the amino acid sequence of SEQ ID NO: 4, a HCDR2 comprising the amino acid sequence of SEQ ID NO: 5, a HCDR3 comprising the amino acid sequence of SEQ ID NO: 6, a light chain complementarity determining region 1 (LCDR1) comprising the amino acid sequence of SEQ ID NO: 13, a LCDR2 comprising the amino acid sequence RAS, and a LCDR3 comprising the amino acid sequence of SEQ ID NO: 15, and wherein the heavy chain further comprises a first CH2 domain, a second CH2 domain, and a third CH2 domain, wherein the first CH2 domain and the second CH2 domain are connected by a first linker and an optional first hinge, and wherein the second CH2 domain and the third CH2 domain are connected by a second linker and an optional second hinge, and wherein the heavy chain further comprises a CH3 domain.Attorney Docket No. TP385350WO1

[0008] Disclosed herein are antibodies that immunospecifically bind to OCT4, wherein the antibodies comprise a heavy chain (HC) and a light chain (LC) that pair to form a binding domain comprising a heavy chain complementarity determining region 1 (HCDR1) comprising the amino acid sequence of SEQ ID NO: 29, a HCDR2 comprising the amino acid sequence of SEQ ID NO: 30, and a HCDR3 comprising the amino acid sequence of SEQ ID NO: 31, a light chain complementarity determining region 1 (LCDR1) comprising the amino acid sequence of SEQ ID NO: 37, a LCDR2 comprising the amino acid sequence GAS, and a LCDR3 comprising the amino acid sequence of SEQ ID NO: 39, and wherein the heavy chain further comprises a first CH2 domain, a second CH2 domain, and a third CH2 domain, wherein the first CH2 domain and the second CH2 domain are connected by a first linker and an optional first hinge, and wherein the second CH2 domain and the third CH2 domain are connected by a second linker and an optional second hinge, and wherein the heavy chain further comprises a CH3 domain.

[0009] Disclosed herein are antibodies that immunospecifically bind to IDO1, wherein the antibodies comprise a heavy chain (HC) and a light chain (LC) that pair to form a binding domain comprising a heavy chain complementarity determining region 1 (HCDR1) comprising the amino acid sequence of SEQ ID NO: 66, a HCDR2 comprising the amino acid sequence of SEQ ID NO: 67, and a HCDR3 comprising the amino acid sequence of SEQ ID NO: 68, a light chain complementarity determining region 1 (LCDR1) comprising the amino acid sequence of SEQ ID NO: 72, a LCDR2 comprising the amino acid sequence of YAS, and a LCDR3 comprising the amino acid sequence of SEQ ID NO: 73, and wherein the heavy chain further comprises a first CH2 domain, a second CH2 domain, and a third CH2 domain, wherein the first CH2 domain and the second CH2 domain are connected by a first linker and an optional first hinge, and wherein the second CH2 domain and the third CH2 domain are connected by a second linker and an optional second hinge, and wherein the heavy chain further comprises a CH3 domain.

[0010] Disclosed herein are antibodies that immunospecifically bind to FLAG-tag, wherein the antibodies comprise a heavy chain (HC) and a light chain (LC) that pair to form a binding domain comprising a heavy chain complementarity determining region 1 (HCDR1) comprising the amino acid sequence of SEQ ID NO: 163, a HCDR2 comprising the amino acid sequence of SEQ ID NO: 164, and a HCDR3 comprising the amino acid sequence of SEQ ID NO: 165, a light chain complementarity determining region 1 (LCDR1) comprisingAttorney Docket No. TP385350WO1 the amino acid sequence of SEQ ID NO: 168, a LCDR2 comprising the amino acid sequence of YAS, and a LCDR3 comprising the amino acid sequence of SEQ ID NO: 169, and wherein the heavy chain further comprises a first CH2 domain, a second CH2 domain, and a third CH2 domain, wherein the first CH2 domain and the second CH2 domain are connected by a first linker and an optional first hinge, and wherein the second CH2 domain and the third CH2 domain are connected by a second linker and an optional second hinge, and wherein the heavy chain further comprises a CH3 domain.

[0011] Disclosed herein are rabbit anti-mouse (RAM) secondary antibodies, wherein the antibodies comprise a heavy chain (HC) and a light chain (LC) that pair to form a binding domain comprising a heavy chain complementarity determining region 1 (HCDR1) comprising the amino acid sequence of SEQ ID NO: 127, a HCDR2 comprising the amino acid sequence of SEQ ID NO: 128, and a HCDR3 comprising the amino acid sequence of SEQ ID NO: 129, a light chain complementarity determining region 1 (LCDR1) comprising the amino acid sequence of SEQ ID NO: 132, a LCDR2 comprising the amino acid sequence of GAS, and a LCDR3 comprising the amino acid sequence of SEQ ID NO: 133, and wherein the heavy chain further comprises a first CH2 domain, a second CH2 domain, and a third CH2 domain, wherein the first CH2 domain and the second CH2 domain are connected by a first linker and an optional first hinge, and wherein the second CH2 domain and the third CH2 domain are connected by a second linker and an optional second hinge, and wherein the heavy chain further comprises a CH3 domain.

[0012] Disclosed herein are antibodies that immunospecifically bind to pERK1 / 2, wherein the antibodies comprise a heavy chain (HC) and a light chain (LC) that pair to form a binding domain comprising a heavy chain complementarity determining region 1 (HCDR1) comprising the amino acid sequence of SEQ ID NO: 85, a HCDR2 comprising the amino acid sequence of SEQ ID NO: 86, and a HCDR3 comprising the amino acid sequence of SEQ ID NO: 87, a light chain complementarity determining region 1 (LCDR1) comprising the amino acid sequence of SEQ ID NO: 91, a LCDR2 comprising the amino acid sequence of KVS, and a LCDR3 comprising the amino acid sequence of SEQ ID NO: 92, and wherein the heavy chain further comprises a first CH2 domain, a second CH2 domain, and a third CH2 domain, wherein the first CH2 domain and the second CH2 domain are connected by a first linker and an optional first hinge, and wherein the second CH2 domain and the third CH2Attorney Docket No. TP385350WO1 domain are connected by a second linker and an optional second hinge, and wherein the heavy chain further comprises a CH3 domain.

[0013] Disclosed herein are antibodies that immunospecifically bind to PSD-95, wherein the antibodies comprise a heavy chain (HC) and a light chain (LC) that pair to form a binding domain comprising a heavy chain complementarity determining region 1 (HCDR1) comprising the amino acid sequence of SEQ ID NO: 111, a HCDR2 comprising the amino acid sequence of SEQ ID NO: 112, and a HCDR3 comprising the amino acid sequence of SEQ ID NO: 113, a light chain complementarity determining region 1 (LCDR1) comprising the amino acid sequence of SEQ ID NO: 116, a LCDR2 comprising the amino acid sequence of DTS, and a LCDR3 comprising the amino acid sequence of SEQ ID NO: 117, and wherein the heavy chain further comprises a first CH2 domain, a second CH2 domain, and a third CH2 domain, wherein the first CH2 domain and the second CH2 domain are connected by a first linker and an optional first hinge, and wherein the second CH2 domain and the third CH2 domain are connected by a second linker and an optional second hinge, and wherein the heavy chain further comprises a CH3 domain.

[0014] Disclosed herein are antibodies that immunospecifically bind to TIGIT, wherein the antibodies comprise a heavy chain (HC) and a light chain (LC) that pair to form a binding domain comprising a heavy chain complementarity determining region 1 (HCDR1) comprising the amino acid sequence of SEQ ID NO: 146, a HCDR2 comprising the amino acid sequence of SEQ ID NO: 147, and a HCDR3 comprising the amino acid sequence of SEQ ID NO: 148, a light chain complementarity determining region 1 (LCDR1) comprising the amino acid sequence of SEQ ID NO: 151, a LCDR2 comprising the amino acid sequence of SAS, and a LCDR3 comprising the amino acid sequence of SEQ ID NO: 152, and wherein the heavy chain further comprises a first CH2 domain, a second CH2 domain, and a third CH2 domain, wherein the first CH2 domain and the second CH2 domain are connected by a first linker and an optional first hinge, and wherein the second CH2 domain and the third CH2 domain are connected by a second linker and an optional second hinge, and wherein the heavy chain further comprises a CH3 domain.

[0015] Disclosed herein are methods of detecting the presence of a target protein in a sample, said methods comprising applying the herein disclosed isolated polypeptides or the herein disclosed antibodies to a sample containing a target protein, incubating the isolated polypeptide or the antibody in the sample under conditions permitting the isolatedAttorney Docket No. TP385350WO1 polypeptides or the polypeptide or the antibody to bind to the target protein and to form a bound isolated polypeptide or a bound polypeptide or the antibody, and determining the presence of the bound isolated polypeptide or the bound polypeptide or the antibody, thereby detecting the presence of the target protein.

[0016] Disclosed herein are methods of increasing the sensitivity of an antibody, comprising introducing into a heavy chain of the antibody a polypeptide comprising the amino acid sequence of Formula I: Z-(X-Yn-Zr)p-X, wherein Z is a hinge, X is a CH2 domain, Y is a linker, n is an integer of 1 or greater, p is an integer of 2 or greater, and r is an integer of 0 or 1, wherein the sensitivity of the antibody is increased at least 1.5-fold compared to a corresponding wild type antibody. In certain embodiments, the method further comprises introducing LFLEPS mutations into the amino acid sequence of X, wherein the LFLEPS mutations comprise L234F, L235E and P331S substitutions.

[0017] Disclosed herein are methods of reducing the background signal of an antibody, comprising introducing into a heavy chain of the antibody a polypeptide comprising the amino acid sequence of Formula I: Z-(X-Yn-Zr)p-X, wherein Z is a hinge, X is a CH2 domain, Y is a linker, n is an integer of 1 or greater, p is an integer of 2 or greater, and r is an integer of 0 or 1, wherein the background signal of the antibody is reduced at least 1.5- fold compared to a corresponding wild type antibody. In certain embodiments, the method further comprises introducing LFLEPS mutations into the amino acid sequence of X, wherein the LFLEPS mutations comprise L234F, L235E and P331S substitutions. BRIEF DESCRIPTION OF THE DRAWINGS

[0018] The foregoing summary, as well as the following detailed description of preferred embodiments of the present application, will be better understood when read in conjunction with the appended drawings. It should be understood, however, that the application is not limited to the precise embodiments shown in the drawings.

[0019] FIG.1A is a schematic diagram of Parent wild type antibody (WT IgG, ~150 kDa)

[0020] FIG.1B is a schematic diagram of the 1TdFc design (IgG-2Fc, ~200 kDa). The 1TdFc design has one additional Fc domain connected by a linker and hinge.

[0021] FIG.1C is a schematic diagram of the 2TdFc design (IgG-3Fc, ~250 kDa). The 2TdFc design has two additional Fc domains connected by a linker and hinge.Attorney Docket No. TP385350WO1

[0022] FIG.1D is a schematic diagram of the 2TdCH2 design (~200 kDa). The 2TdCH2 design has two additional CH2 domains connected by linker and optional hinge.

[0023] FIG.1E is a schematic diagram of the 2TdCH2-LFLEPS design (~200 kDa). The 2TdCH2-LFLEPS design has two additional CH2 domains connected by a linker and optional hinge, and FcR mutations (stars) represented by the acronym “LFLEPS”.

[0024] FIG.2A is a plasmid map showing that the polynucleotide sequence of the anti-OCT4 antibody heavy chain (HC) with the 2TdCH2-LFLEPS configuration was synthesized and cloned between XbaI-NotI restriction sites in pcDNA3.4 Vector. XbaI-KasI restriction enzyme can be used for subcloning of another target HC “Fab” region.

[0025] FIG.2B is a plasmid map showing that the polynucleotide sequence of the anti-OCT4 antibody light chain (LC) was synthesized and cloned between XbaI-NotI restriction sites in pcDNA3.4 Vector. XbaI-NotI restriction enzyme can be used for subcloning of another target LC “Fab” region.

[0026] FIG.2C shows a restriction digestion analysis of the anti-OCT4 antibody HC with the 2TdCH2-LFLEPS configuration and anti-OCT4 antibody LC. Anti-OCT4 antibody HC 2TdCH2-LFLEPS shows a 2203 bp insert released upon XbaI-NotI digestion. Anti-OCT4 antibody LC shows a 746 bp insert released upon XbaI-NotI digestion.

[0027] FIG.3A is a plasmid map showing that the polynucleotide sequence of the anti-Parkin antibody heavy chain (HC) with 2TdCH2-LFLEPS configuration was synthesized and cloned between XbaI-NotI restriction sites in pcDNA3.4 Vector. XbaI-KasI restriction enzyme can be used for subcloning of another target HC “Fab” region.

[0028] FIG.3B is a plasmid map showing that the polynucleotide sequence of the anti-Parkin antibody light chain (LC) was synthesized and cloned between XbaI-NotI restriction sites in pcDNA3.4 Vector. XbaI-NotI restriction enzyme can be used for subcloning of another target LC “Fab” region.

[0029] FIG.3C shows a restriction digestion analysis of anti-Parkin antibody HC with the 2TdCH2-LFLEPS configuration and anti-Parkin antibody LC. Anti-Parkin antibody HC 2TdCH2-LFLEPS shows a 2218 bp insert released upon XbaI-NotI digestion. Anti- Parkin antibody LC shows a 743 bp insert released upon XbaI-NotI digestion.

[0030] FIG.4A shows a representative HPLC profile of parent anti-Parkin wild type (WT) antibody having purity of 95% monomer.Attorney Docket No. TP385350WO1

[0031] FIG.4B shows a representative HPLC profile of anti-OCT41TdFc antibody having purity of 63% monomer.

[0032] FIG.4C shows a representative HPLC profile of anti-Parkin 2TdFc antibody having purity of 33% monomer.

[0033] FIG.4D shows a representative HPLC profile of anti-Parkin 2TdCH2- LFLEPS antibody having purity of 87% monomer.

[0034] FIG.5A is a western blot (WB) analysis performed on whole cell extracts of rat brain tissue (left panel) and PC-12 cells (right panel). The blots were probed with wild type (WT) and engineered anti-Parkin antibodies (1TdFC, 2TdFc, 2TdCH2). A ~48 kDa band corresponding to Parkin was observed in rat brain and PC-12 cell lysates tested.

[0035] FIG.5B is a western blot (top panel) showing how antibody specificity was demonstrated by siRNA-mediated knockdown of the target protein. SH-SY5Y cells were transfected with Parkin siRNA and a decrease in signal intensity was observed in western blot application using anti-Parkin 2TdCH2-LFLEPS antibody. The bottom panel is a graphical representation of the western blot analysis.

[0036] FIG.5C is a western blot analysis that was performed on whole cell extracts of PC-12 cells. The blots were probed with wild type (WT) and engineered anti-Parkin antibodies (2TdCH2, 2TdCH2-LFLEPS). A similar intensity of ~48 kDa band corresponding to Parkin was observed in PC-12 cell lysates with 2TdCH2 vs 2TdCH2-LFLEPS.

[0037] FIG.5D is a scatter plot analysis of the WB data: anti-Parkin 2TdCH2 antibody shows significant fold enhancement (**p<0.01) compared to WT antibody.

[0038] FIG.6A is a western blot analysis that was performed on whole cell extracts of NTERA-2 (left panel) and F9 cells (right panel). The blots were probed with wild type (WT) and engineered anti-OCT4 antibodies (1TdFC, 2TdFc, 2TdCH2). A ~38 kDa band corresponding to OCT4 was observed in the cell lysates tested.

[0039] FIG.6B is a western blot analysis performed on whole cell extracts of NTERA-2 cells. The blots were probed with wild type (WT) and engineered anti-OCT4 antibodies (2TdCH2, 2TdCH2-LFLEPS). A similar intensity of ~38 kDa band corresponding to OCT4 was observed in NTERA-2 cell lysates with anti-OCT42TdCH2 vs 2TdCH2- LFLEPS antibodies.

[0040] FIG.6C shows antibody specificity was demonstrated by relative expression (top panel) with BOI in NTERA-2, NCCIT and F9 cells. HEK293, A431 and HELA wereAttorney Docket No. TP385350WO1 negative cell lines. siRNA-mediated knockdown (bottom panel) of the target protein. NTERA-2 cells were transfected with OCT4 siRNA and a decrease in signal intensity was observed in western blot application using anti-OCT42TdCH2-LFLEPS antibody.

[0041] FIG.6D is a scatter plot analysis of WB data: anti-OCT42TdCH2 antibody show significant fold enhancement (****p<0.0001) compared to WT antibody.

[0042] FIG.7A is a bar graph analysis of western blot data obtained from anti- Parkin 2TdCH2 antibody with various secondary antibodies as indicated. The anti-Parkin 2TdCH2 antibody showed fold enhancement across different secondaries compared to wild type (WT) antibody.

[0043] FIG.7B is a bar graph analysis of western blot data obtained from anti- OCT4 2TdCH2 antibody with various secondary antibodies as indicated. The anti-OCT4 2TdCH2 antibody showed fold enhancement across different secondaries compared to wild type (WT) antibody.

[0044] FIG.8A is an immunofluorescent analysis of OCT4 performed using 70% confluent NTERA-2 cells. The cells were stained with different concentrations of anti-OCT4 wild type (WT) antibody (upper panel) or anti-OCT42TdCH2-LFLEPS antibody (1:1,000) (lower panel).

[0045] FIG.8B shows a knockdown of OCT4 was achieved by transfecting NTERA-2 cells with specific validated siRNA (Silencer® select Product # s10871, s10873). Immunofluorescence analysis was performed on NTERA-2 cells (untransfected, panels a-d), transfected with OCT4 specific siRNA (panel i-l) or non-specific scrambled siRNA (panels e- h).

[0046] FIG.9A is an immunohistochemical analysis of Parkin performed using formalin-fixed paraffin-embedded mouse kidney tissue sections. The sections were probed with anti-Parkin wild type (WT) (lower panel) or anti-Parkin 2TdCH2-LFLEPS (upper panel) (1:100 dilution) antibody.

[0047] FIG.9B is an immunohistochemical analysis of Parkin performed using formalin-fixed paraffin-embedded mouse kidney tissue sections. The sections were probed with anti-Parkin polyclonal antibody (Benchmark (BM) or positive control) (lower panel) or Parkin 2TdCH2-LFLEPS (upper panel) (1:100 dilution) antibody.

[0048] FIG.10 is a flow cytometry analysis of Parkin performed in PC-12 cells. Cells were fixed and permeabilized using the Intracellular Fixation & PermeabilizationAttorney Docket No. TP385350WO1 Buffer Set (Product # 88-8824-00) and then stained intracellularly with 2 µg of anti-Parkin wild type (WT) antibody or anti-Parkin 2TdCH2-LFLEPS antibody or positive control (PC) antibody (PA5-1339) antibody. Unstained samples were used as a control. The top panel shows the flow cytometry spectra and the bottom panel shows a graphical representation of the flow data.

[0049] FIG.11 is a graphical representation of an immunofluorescent analysis of IDO1 that was performed using 70% confluent CD64-GFP (FcγR1) transfected HEK-293E cells demonstrating the amount of FcγR1 binding by the antibodies. The cells were stained with 0.5 µg / ml and 2.5 µg / ml of IDO1 wild type (WT) ± LFLEPS antibody or anti-IDO1 2TdCH2 ± LFLEPS antibody followed by Goat anti-Rabbit Alexa Fluor™ 647 secondary antibody (Product #A21244).

[0050] FIG.12A is a flow cytometry analysis of IDO1 that was performed using 70% confluent CD64-GFP (FcγR1) transfected HEK-293E cells. The cells were stained with 0.5 µg / ml (lower panel) and 2.5 µg / ml (upper panel) with anti-IDO1 wild type ± LFLEPS (WT) antibody or anti-IDO12TdCH2 ± LFLEPS antibody followed by Goat anti-Rabbit Alexa Fluor™ 647 secondary antibody (Product #A21244) Viable cells were used for analysis and data was acquired using Attune™ NxT Flow Cytometer (Product # A29004). A shift in the histogram towards the left represents the background signal reduction upon LFLEPS mutation.

[0051] FIG.12B is a graphical representation of the flow cytometry analysis shown in FIG.12A demonstrating the amount of FcγR1 binding by the antibodies. The asterisk (*) indicates statistical significance as determined by p<0.05 using 2-tailed Student’s T test.

[0052] FIG.13A is a graphical representation of an immunofluorescent analysis of IDO1 that was performed using 70% confluent CD64-GFP (FcγR1) transfected HEK-293E cells with / without treatment of Interferon-gamma (INFγ). The cells were stained with 2.5 µg / ml of anti-IDO1 wild type (WT) antibody ± LFLEPS or anti-IDO12TdCH2 ± LFLEPS antibody, followed by Goat anti-Rabbit Alexa Fluor™ 647 secondary antibody (Product #A21244).

[0053] FIG.13B is a graphical representation of an immunofluorescent analysis of IDO1 that was performed using 70% confluent CD64-GFP (FcγR1) transfected HEK-293E cells with / without treatment of INFγ. The cells were stained with 0.5 µg / ml of anti-IDO1Attorney Docket No. TP385350WO1 wild type (WT) antibody ± LFLEPS or anti-IDO12TdCH2 ± LFLEPS antibody, followed by Goat anti-Rabbit Alexa Fluor™ 647 secondary antibody (Product #A21244).

[0054] FIG.14A is a flow cytometry analysis demonstrating that the 2TdCH2- LFLEPS design in a mouse-rabbit chimeric antibody (Fab-Fc Chimera) enhances TIGIT signal detection in human PBMCs compared to wild type design. Human PBMCs were stained with CD4 FITC (Product # 11-0049-42) and 0.25 µg of anti-TIGIT wild type antibody, or anti-TIGIT + LFLEPS antibody, or anti-TIGIT + 2TdCH2-LFLEPS antibody, followed by Goat anti-Rabbit IgG (H+L) Alexa Fluor™ Plus 647 secondary antibody (Product # A55060). Viable cells in the lymphocyte gate were used for analysis and data was acquired using Attune™ NxT Flow Cytometer (Product # A29004).

[0055] FIG.14B is a graphical representation of the flow cytometry analysis shown in FIG.14A showing the staining index of the various antibodies. The asterisk (*) indicates statistical significance as determined by p<0.05 and asterisk (**) indicates statistical significance as determined by p<0.01 using 2-tailed Student’s T test.

[0056] FIG.15 is a graphical representation of an immunofluorescent analysis of the effect of the 2TdCH2 LFLEPS design on a rabbit anti-mouse (RAM) IgG secondary antibody and showing the staining index of the various antibodies. The analysis was performed using 70% confluent CD64-GFP (FcγR1) transfected HEK-293E cells. The cells were stained with 0.5 µg / ml and 2.5 µg / ml of a RAM wild type (WT) antibody ± LFLEPS or a RAM 2TdCH2-LFLEPS ± LFLEPS antibody followed by Donkey anti-Rabbit Alexa Fluor™ 647 secondary antibody d(A32795) to detect the RAM antibody. Increased staining with the RAM WT antibody and the RAM 2TdCH2 antibody (without LFLEPS) indicates the non-specific binding of the secondary antibody as there is no primary antibody present in the sample. The asterisk (*) indicates statistical significance as determined by p<0.05 using 2- tailed Student’s T test.

[0057] FIG.16A shows a western blot (left panel) that was performed using 0.5 µg / ml of anti-pERK1 / 2 WT antibody and anti-pERK1 / 22TdCH2 recombinant mouse monoclonal antibody followed by Goat anti-Mouse IgG (H+L) Horseradish Peroxidase (HRP) Superclonal™ secondary antibody (Product # A28177) by loading whole cell extracts of Jurkat cells with and without PMA treatment (200 ng / ml). The right panel shows a graphical representation of the western blot analysis.Attorney Docket No. TP385350WO1

[0058] FIG.16B is an immunofluorescent analysis was performed using 2.5 µg / ml of anti-pERK1 / 2 WT antibody and anti-pERK1 / 22TdCH2 recombinant mouse monoclonal antibody followed by Goat anti-Mouse IgG (H+L) Alexa Fluor™ Plus 488 secondary antibody in serum starved A431 cells with EGF treatment (200 ng / ml). The left panel shows the immunofluorescence images and the right panel shows a graphical representation of the immunofluorescent analysis.

[0059] FIG.17 shows a western blot that was performed using 4.0 to 0.5 µg / ml of anti-PSD-95 WT antibody and anti-PSD-952TdCH2 antibody followed by Goat anti-Mouse IgG (H+L) HRP Superclonal™ Secondary antibody (Product # A28177) by loading whole cell extracts of Neuro2A cells.

[0060] FIG.18A is an immunofluorescent analysis that was performed using 1 µg / ml of wild type or 2TdCH2 LFLEPS anti-OCT4-Alexa Fluor™ Plus 555 antibody conjugate in NTERA2 cells.

[0061] FIG.18B is a graphical representation of the immunofluorescent analysis shown in FIG.18A. The asterisk (**) indicates statistical significance as determined by p<0.01 using 2-tailed Student’s T test.

[0062] FIG.19A is an immunofluorescent analysis that was performed using 1 µg / ml of wild type or 2TdCH2 LFLEPS anti-OCT4-Alexa Fluor™ Plus 647 conjugate in NTERA2 cells.

[0063] FIG.19B is a graphical representation of the immunofluorescent analysis shown in FIG.19A. The asterisk (*) indicates statistical significance as determined by p<0.05 using 2-tailed Student’s T test.

[0064] FIG.20A is western blot analysis that was performed using 1 µg / ml of wild type or 2TdCH2 LFLEPS anti-Parkin HRP conjugate at 4 and 8 molar ratio in PC-12 cell whole cell lysate (WCL).

[0065] FIG.20B is western blot analysis that was performed using 1 µg / ml of wild type or 2TdCH2 LFLEPS anti-FLAG-tag HRP conjugate at 4 and 8 molar ratio in EXPI293 cell transfected with FLAG-Histone H4 construct.

[0066] FIG.20C is western blot analysis that was performed using 1 µg / ml of wild type or 2TdCH2 LFLEPS anti-FLAG-tag Biotin conjugate at 25 and 50 molar ratio in EXPI293 cell transfected with FLAG-Histone H4 construct.Attorney Docket No. TP385350WO1

[0067] FIG.21A indicates degree of labeling (DOL) from SiteClick™ conjugation of Alexa Fluor™ 488 dyes with anti-Parkin WT, 2TdCH2 and 2TdCH2 LFLEPS antibodies. The asterisk (*) indicates statistical significance as determined by p<0.05 and asterisk (**) indicates statistical significance as determined by p<0.01 using 2-tailed Student’s T test.

[0068] FIG.21B indicates degree of labeling (DOL) from SiteClick™ conjugation of Alexa Fluor™ 488 dyes with anti-OCT4 WT, 2TdCH2 and 2TdCH2 LFLEPS antibodies. The asterisk (*) indicates statistical significance as determined by p<0.05 using 2- tailed Student’s T test. DETAILED DESCRIPTION

[0069] Various publications, articles and patents are cited or described in the background and throughout the specification; each of these references is herein incorporated by reference in its entirety. Discussion of documents, acts, materials, devices, articles or the like which has been included in the present specification is for the purpose of providing context for the invention. Such discussion is not an admission that any or all of these matters form part of the prior art with respect to any inventions disclosed or claimed.

[0070] The disclosed polypeptides, antibodies and methods may be understood more readily by reference to the following detailed description taken in connection with the accompanying figures, which form a part of this disclosure. It is to be understood that the disclosed polypeptides, antibodies, and methods are not limited to the specific polypeptides, antibodies, and methods described and / or shown herein, and that the terminology used herein is for the purpose of describing particular embodiments by way of example only and is not intended to be limiting of the claimed polypeptides, antibodies, and methods.

[0071] Unless specifically stated otherwise, any description as to a possible mechanism or mode of action or reason for improvement is meant to be illustrative only, and the disclosed polypeptides, antibodies, and methods are not to be constrained by the correctness or incorrectness of any such suggested mechanism or mode of action or reason for improvement.

[0072] Throughout this text, the descriptions refer to polypeptides, antibodies, and methods of producing the polypeptides and antibodies. Where the disclosure describes or claims a feature or embodiment associated with polypeptides or antibodies, such a feature or embodiment is equally applicable to the methods of producing the polypeptides or antibodies.Attorney Docket No. TP385350WO1 Likewise, where the disclosure describes or claims a feature or embodiment associated with methods of producing the polypeptide or antibodies, such a feature or embodiment is equally applicable to the polypeptides or antibodies.

[0073] It is to be appreciated that certain features of the disclosed polypeptides, antibodies, and methods which are, for clarity, described herein in the context of separate embodiments, may also be provided in combination in a single embodiment. Conversely, various features of the disclosed polypeptides, antibodies, and methods that are, for brevity, described in the context of a single embodiment, may also be provided separately or in any subcombination.

[0074] As used herein, the singular forms “a,” “an,” and “the” include the plural.

[0075] When a list is presented, unless stated otherwise, it is to be understood that each individual element of that list, and every combination of that list, is a separate embodiment. For example, a list of embodiments presented as “A, B, or C” is to be interpreted as including the embodiments: “A;” “B;” “C;” “A or B;” “A or C;” “B or C;” or “A, B, or C.”

[0076] Various terms relating to aspects of the description are used throughout the specification and claims. Such terms are to be given their ordinary meaning in the art unless otherwise indicated. Other specifically defined terms are to be construed in a manner consistent with the definitions provided herein.

[0077] The term “comprising” is intended to include examples encompassed by the terms “consisting essentially of” and “consisting of”; similarly, the term “consisting essentially of” is intended to include examples encompassed by the term “consisting of.” Certain Terminology

[0078] “Antibody” as used herein means an antibody of classes IgG, IgM, IgA, IgD, or IgE, or fragments, or derivatives thereof, derived from any animal including, but not limited to, sheep, rabbits, goats, mice, camelids, and egg yolk.

[0079] Antibodies include but are not limited to monoclonal or polyclonal antibodies, bispecific antibodies, bifunctional antibodies, fragments of antibodies, humanized antibodies, chimeric antibodies, engineered antibodies, antibody constructs, and altered antibody-like structures. Engineered antibodies or antibody constructs include antibody-like structures having the herein described polypeptides comprising the amino acid sequence ofAttorney Docket No. TP385350WO1 Formula I: Z-(X-Yn-Zr)p-X, wherein Z is a hinge comprising an amino acid sequence at least 90% identical to SEQ ID NO: 18, SEQ ID NO: 48, or SEQ ID NO: 102, X is a CH2 domain comprising an amino acid sequence at least 90% identical to SEQ ID NO: 19, SEQ ID NO: 49, or SEQ ID NO: 104, Y is a linker, n is an integer of 1 or greater, p is an integer of 2 or greater, and r is an integer of 0 or 1. Antibodies may be produced or modified by recombinant DNA or other synthetic techniques. The antibody can be an antibody isolated from the serum sample of a mammal, an affinity purified antibody, or any mixture thereof which exhibits sufficient binding specificity to a desired epitope or a sequence derived therefrom. Also included are functional derivatives or “equivalents” of antibodies, e.g., single chain antibodies, CDR-grafted antibodies, etc. Also included are VHH antibodies that may be monovalent or bivalent.

[0080] “Biotin” as used herein means biotin or any biotin derivative, including without limitation, substituted and unsubstituted biotin, and analogs and derivatives thereof, as well as substituted and unsubstituted derivatives of caproylamidobiotin, biocytin, desthiobiotin, desthiobiocytin, iminobiotin, and biotin sulfone.

[0081] “Coding sequence” or “polynucleotide encoding” as used herein means RNA or DNA molecules that comprise a nucleotide sequence which encodes a protein. The coding sequence can further include initiation and termination signals operably linked to regulatory elements including a promoter and polyadenylation signal capable of directing expression in the cells of a subject or mammal to which the nucleic acid molecule is administered.

[0082] “Identity” or “percent identity” as used herein in the context of two or more nucleic acid molecules or polypeptide sequences means that the sequences have a specified percentage of residues that are the same over a specified region. The percentage can be calculated by optimally aligning the two sequences, comparing the two sequences over the specified region, determining the number of positions at which the identical residue occurs in both sequences to yield the number of matched positions, dividing the number of matched positions by the total number of positions in the specified region, and multiplying the result by 100 to yield the percentage of sequence identity. In cases where the two sequences are of different lengths or the alignment produces one or more staggered ends and the specified region of comparison includes only a single sequence, the residues of single sequence are included in the denominator but not the numerator of the calculation. When comparing DNA and RNA, thymine (T) and uracil (U) can be considered equivalent. Identity can beAttorney Docket No. TP385350WO1 performed manually or by using a computer sequence algorithm such as BLAST or BLAST 2.0.

[0083] “Fluorophore” as used herein means a chemical compound that, when excited by exposure to a particular wavelength of light, emits light (fluoresces), for example at a different wavelength than that to which it was exposed. Fluorophores may contain substituents that alter the solubility, spectral properties or physical properties of the fluorophore.

[0084] Exemplary fluorophores include, but are not limited to, xanthene, fluorescein, rhodamine, silicon rhodamine, rhodol, roseamine, carbopyranone, indole, indacene, borapolyazaindacene, furan, benzofuran, cyanine, benzocyanine, benzopyrilium, pyrene, coumarin, styryl, squarine, resorufin, anthraquinone, acridine, benzophenoxazine, quinoline, quinazolinone, oligonucleotide-based fluorescent dyes, conjugated polymer dyes, and tandem dyes.

[0085] Examples of fluorophores suitable for use with the disclosed assays, include, but are not limited to, 6-carboxyfluorescein (FAM), tetrachlorofluorescein (TET), tetramethylrhodamine (TMR), hexachlorofluorescein (HEX), JOE, 6-carboxy-X-rhodamine (ROX), CAL FluorTM, PulsarTM, QuasarTM, Texas RedTM, Texas Red-X, CyTM3, CyTM5, BODIPYTM(boron-dipyrromethene), Alexa FluorTMdyes (such as Alexa FluorTM488), Alexa Fluor Plus dyes (such as Alexa Fluor™ Plus 488), DyLight™ dyes (such as DyLight™ 488), Oregon GreenTM, and NovaFluor™ dyes (such as NovaFluor™ Red 710), as well as other fluorophores described in Johnson, I. and Spence, M.T.Z, (Eds.). (2010) The Molecular Probes Handbook: A Guide to Fluorescent Probes and Labeling Technologies, 11thedition.

[0086] “Immunospecifically binds” as used herein refers to the ability of the disclosed polypeptides or antibodies to preferentially bind to their target(s) without preferentially binding other molecules in a sample or environment containing a mixed population of molecules.

[0087] “Label” as used herein means an agent capable of detection, for example by ELISA, spectrophotometry, flow cytometry, or microscopy. For example, a label can be attached (e.g., covalently linked or conjugated) to a nucleic acid molecule or protein (such as an antibody or antigen binding fragment thereof), thereby permitting detection of the nucleic acid molecule or protein. Examples of labels include, but are not limited to, radioactive isotopes, enzyme substrates, co-factors, ligands, chemiluminescent agents, fluorophores,Attorney Docket No. TP385350WO1 haptens, enzymes, and combinations thereof. Specific non-limiting examples of labels include fluorescent and fluorogenic moieties (e.g., fluorophores).

[0088] Illustrative labels include labels that can be directly observed or measured or indirectly observed or measured. Such labels include, but are not limited to, radiolabels that can be measured with radiation-counting devices; pigments, dyes or other chromogens that can be visually observed or measured with a spectrophotometer; spin labels that can be measured with a spin label analyzer; and fluorescent moieties, where the output signal is generated by the excitation of a suitable molecular adduct and that can be visualized by excitation with light that is absorbed by the dye or can be measured with standard fluorometers or imaging systems, for example.

[0089] The label can be a luminescent substance such as a phosphor or fluorogen; a bioluminescent substance; a chemiluminescent substance, where the output signal is generated by chemical modification of the signal compound; a metal-containing substance; or an enzyme, where there occurs an enzyme-dependent secondary generation of signal, such as the formation of a colored product from a colorless substrate. The label may also take the form of a chemical or biochemical, or an inert particle, including but not limited to colloidal gold, microspheres, quantum dots, or inorganic crystals such as nanocrystals or phosphors (see, e.g., Beverloo, et al., Anal. Biochem.203, 326-34 (1992)).

[0090] The term label can also refer to a “tag” or hapten that can bind selectively to a labeled molecule such that the labeled molecule, when added subsequently, is used to generate a detectable signal. For instance, one can use biotin, iminobiotin or desthiobiotin as a tag and then use an avidin or streptavidin conjugate of horseradish peroxidase (HRP) to bind to the tag, and then use a chromogenic substrate (e.g., tetramethylbenzidine) or a fluorogenic substrate such as Amplex™ Red or Amplex™ Gold (Thermo Fisher Scientific, Waltham, MA) to detect the presence of HRP. In a similar fashion, the tag can be a hapten or antigen (e.g., digoxigenin), and an enzymatically, fluorescently, or radioactively labeled antibody can be used to bind to the tag.

[0091] Numerous labels are known by those of skill in the art and include, but are not limited to, particles, fluorescent dyes, haptens, enzymes and their chromogenic, fluorogenic, and chemiluminescent substrates, and other labels that are described in Johnson, I. and Spence, M.T.Z, (Eds.). (2010) The Molecular Probes Handbook: A Guide to Fluorescent Probes and Labeling Technologies, 11thedition.Attorney Docket No. TP385350WO1

[0092] “Nucleic acid molecule,” “oligonucleotide,” or “polynucleotide” as used herein means at least two nucleotides covalently linked together. The depiction of a single strand also defines the sequence of the complementary strand. Thus, a nucleic acid molecule also encompasses the complementary strand of a depicted single strand. Many variants of a nucleic acid molecule can be used for the same purpose as a given nucleic acid molecule. Thus, a nucleic acid molecule also encompasses substantially identical nucleic acid molecules and complements thereof.

[0093] Nucleic acid molecules can be single stranded or double stranded, or can contain portions of double stranded and single stranded sequence. The nucleic acid molecule can be DNA (both genomic and cDNA), RNA, or a hybrid, where the nucleic acid molecule can contain combinations of deoxyribo- and ribo-nucleotides, and combinations of bases including uracil, adenine, thymine, cytosine, guanine, inosine, xanthine hypoxanthine, isocytosine and isoguanine. Nucleic acid molecules can have post-transcriptional chemical modifications. Nucleic acid molecules can be obtained by chemical synthesis methods or by recombinant methods.

[0094] “Oligonucleotide-based fluorescent dye” as used herein means a nucleic acid nanostructure composed of a DNA-based scaffold that precisely arranges fluorophores in order to engineer their interactions and the overall fluorescent properties of the structure. Examples of oligonucleotide-based fluorescent dyes are described in International Patent Application Publication No. WO 2018 / 231805 and include, but are not limited to, NovaFluor™ dyes (Thermo Fisher Scientific, Waltham, MA).

[0095] A “peptide,” “protein,” or “polypeptide” as used herein can mean a linked sequence of amino acids and can be natural, synthetic, or a modification or combination of natural and synthetic. A peptide, protein, or polypeptide can have post-translational modifications.

[0096] “Vector” as used herein means a polynucleotide sequence containing an origin of replication. A vector can be a viral vector, bacteriophage, bacterial artificial chromosome, or yeast artificial chromosome. A vector can be a DNA or RNA vector. A vector can be a self-replicating extrachromosomal vector, and preferably, is a DNA plasmid. The vector can contain or include one or more heterologous polynucleotide sequences. Engineered AntibodiesAttorney Docket No. TP385350WO1

[0097] Recombinant antibodies offer the ability to engineer benefits in a rational method. One of the most important characteristics of an antibody is its sensitivity, i.e. the strength of the signal produced in a particular immunoassay. A highly sensitive antibody has several benefits, such as the detection of low abundance or rare targets and the ability to detect the protein of interest in complex assays such as imaging. Antibody sensitivity is influenced both by increased signal and decreased background. Antibody sensitivity can be increased by affinity maturation of the antibody clone, which involves mutations of the antigen binding site or paratope. However, affinity maturation has to be tailored for each antibody and therefore is not a scalable solution to address a large number of antibodies. Furthermore, mutations to the paratope change the clone of the antibody and essentially produces a new antibody.

[0098] The instant disclosure provides an approach for increasing antibody sensitivity by rational modification of the antibody Fc region. Amplification or expansion of the entire Fc domain or portions of the Fc domain provides additional binding sites for secondary antibodies, resulting in an enhanced signal. Prior studies have utilized Fc amplification for other purposes such as increased antibody effects in a therapeutic application by enhanced binding to Fc receptors, in the case of a humanized antibody. It was found that antibody modification by adding one additional Fc domain (1TdFc) or two additional Fc domains (2TdFc) to the antibody resulted in poor expression resulting in a low yield and have aggregation and purity challenges. To overcome the above limitations, the instant disclosure provides a rational design to circumvent the disadvantages of the prior studies by selectively adding additional CH2 domains (2TdCH2) instead of the entire Fc region. The 2TdCH2 design of the instant polypeptides and antibodies provided herein overcame the above challenges and it was unexpectedly found that the 2TdCH2 engineered antibodies provided herein had comparable yields, monomer purity, and stability to wildtype antibodies. The two additional CH2 domains, joined by a hinge and a linker, offer additional epitopes for secondary antibody binding and additional lysine residues for conjugation. In addition, the 2TdCH2 engineered antibodies of the present disclosure provide a significant increase in sensitivity in multiple applications such as western blotting and immunocytochemistry and allow for broader applicability of the antibodies. In addition to the benefits for the unconjugated antibodies, 2TdCH2 engineering also provides additional binding sites for different conjugation chemistries. It was also unexpectedly found thatAttorney Docket No. TP385350WO1 introduction of specific mutations, L234F, L235E and P331S, herein described as “LFLEPS mutations”, into the engineered backbone reduced background binding of the antibody to Fc gamma receptors present on selected cells. Thus, the advantages of the engineered antibodies of the present disclosure include: i) increased sensitivity of at least 1.5-fold; ii) reduced background by at least 1.5-fold; ii) detection of low abundance targets that are otherwise difficult to detect with standard antibodies; iii) increased application coverage through the use of multiplexing; iv) increased conjugation sites for primary conjugates; and v) addresses design challenges of expression, purity and stability found with other recombinant antibodies.

[0099] Disclosed herein are isolated polypeptides comprising, consisting essentially of, or consisting of the amino acid sequence of Formula I: Z-(X-Yn-Zr)p-X, wherein: Z is a hinge region, X is a CH2 domain, Y is a linker, n is an integer of 1 or greater, p is an integer of 2 or greater, and r is an integer of 0 or 1. In certain embodiments the elements of the amino acid sequence of Formula I defined herein as Z-(X-Yn-Zr)p-X are linked directly to one another with no intervening amino acid sequences.

[0100] The hinge (Z) region can comprise the amino acid sequence of SEQ ID NO: 18. The hinge (Z) can comprise an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 18. The hinge (Z) can comprise an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 18. The hinge (Z) can comprise the amino acid sequence of SEQ ID NO: 48. The hinge (Z) can comprise an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 48. The hinge (Z) can comprise an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 48. The hinge (Z) can comprise the amino acid sequence of SEQ ID NO: 102. The hinge (Z) can comprise an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 102. The hinge (Z) can comprise an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 102.

[0101] The CH2 domain (X) can comprise the amino acid sequence of SEQ ID NO: 19. The CH2 domain (X) can comprise an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 19. The CH2 domain (X) can comprise an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 19. The CH2 domain (X) can comprise the amino acid sequence of SEQ ID NO: 49. The CH2 domain (X) can comprise an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 49. The CH2 domain (X) can comprise an amino acid sequence with at least 95% sequence identity to SEQAttorney Docket No. TP385350WO1 ID NO: 49. The CH2 domain (X) can comprise the amino acid sequence of SEQ ID NO: 104. The CH2 domain (X) can comprise an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 104. The CH2 domain (X) can comprise an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 104.

[0102] The isolated polypeptide can comprise one or more linkers (Y) comprising the amino acid sequence of GS, GGS, SEQ ID NO: 14, SEQ ID NO: 38, SEQ ID NO: 43, or SEQ ID NO: 44. The one or more linkers (Y) can comprise GS. The one or more linkers (Y) can comprise GGS. The one or more linkers (Y) can comprise SEQ ID NO: 14. The one or more linkers (Y) can comprise SEQ ID NO: 38. The one or more linkers can comprise SEQ ID NO: 43. The one or more linkers (Y) can comprise a sequence with at least 90% sequence identity to SEQ ID NO: 43. The one or more linkers (Y) can comprise SEQ ID NO: 44. The one or more linkers (Y) can comprise a sequence with at least 90% sequence identity to SEQ ID NO: 44.

[0103] The isolated polypeptide can comprise one linker (Y), such that n is 1. The isolated polypeptide can comprise two linkers (Y), such that n is 2. The isolated polypeptide can comprise three linkers (Y), such that n is 3. The isolated polypeptide can comprise four linkers (Y), such that n is 4. The isolated polypeptide can comprise five linkers (Y), such that n is 5.

[0104] The isolated polypeptide can comprise two or more regions comprising a CH2-linker region ((X-Yn)p), such that p is 2 or greater. The isolated polypeptide can comprise two CH2-linker regions ((X-Yn)p), such that p is 2. The isolated polypeptide can comprise three CH2-linker regions ((X-Yn)p), such that p is 3. The isolated polypeptide can comprise four CH2-linker regions ((X-Yn)p), such that p is 4. The isolated polypeptide can comprise five CH2-linker regions ((X-Yn)p), such that p is 5.

[0105] The isolated polypeptide can comprise two or more regions comprising a CH2-linker-hinge region ((X-Yn-Zr)p), such that p is 2 or greater. The isolated polypeptide can comprise two CH2-linker-hinge regions ((X-Yn-Zr)p), such that p is 2. The isolated polypeptide can comprise three CH2-linker-hinge regions ((X-Yn-Zr)p), such that p is 3. The isolated polypeptide can comprise four CH2-linker-hinge regions ((X-Yn-Zr)p), such that p is 4. The isolated polypeptide can comprise five CH2-linker-hinge regions ((X-Yn-Zr)p), such that p is 5.Attorney Docket No. TP385350WO1

[0106] In some embodiments, the isolated polypeptide comprises, consists essentially of, or consists of (X-Yn-Zr)p, wherein Z (hinge) comprises the amino acid sequence of SEQ ID NO: 18, X (CH2 domain) comprises the amino acid sequence of SEQ ID NO: 19, Y (linker) comprises the amino acid sequence of SEQ ID NO: 44, wherein n is 1, r is 0 or 1, and p is 2.

[0107] In some embodiments, the isolated polypeptide comprises, consists essentially of, or consists of (X-Yn-Zr)p, wherein Z (hinge) comprises the amino acid sequence of SEQ ID NO: 48, X (CH2 domain) comprises the amino acid sequence of SEQ ID NO: 49, Y (linker) comprises the amino acid sequence of SEQ ID NO: 44, wherein n is 1, r is 0 or 1, and p is 2.

[0108] In some embodiments, the isolated polypeptide comprises, consists essentially of, or consists of (X-Yn-Zr)p, wherein Z (hinge) comprises the amino acid sequence of SEQ ID NO: 102, X (CH2 domain) comprises the amino acid sequence of SEQ ID NO: 104, Y (linker) comprises the amino acid sequence of SEQ ID NO: 44, wherein n is 1, r is 0 or 1, and p is 2.

[0109] In some embodiments, the isolated polypeptide comprises, consists essentially of, or consists of (X-Yn-Zr)p, wherein Z (hinge) comprises the amino acid sequence of SEQ ID NO: 102, X (CH2 domain) comprises the amino acid sequence of SEQ ID NO: 104, Y (linker) comprises the amino acid sequence of SEQ ID NO: 45, wherein n is 1, r is 0 or 1, and p is 2.

[0110] In some embodiments, the isolated polypeptide comprises two additional CH2 domains N-terminal to the Fc domain connected via a hinge and optionally, one or more linkers. The isolated polypeptide comprising two additional CH2 domains N-terminal to the Fc domain connected via a hinge and optionally one or more linkers can comprise the amino acid sequence of SEQ ID NO: 52 or SEQ ID NO: 54. In some embodiments, the isolated polypeptide comprising a CH3 domain comprises SEQ ID NO: 52. The isolated polypeptide comprising two additional CH2 domains N-terminal to the Fc domain connected via a hinge and optionally one or more linkers can comprise an amino acid sequence with at least 90% identity to SEQ ID NO: 52. The isolated polypeptide comprising two additional CH2 domains N-terminal to the Fc domain connected via a hinge and optionally one or more linkers can comprise an amino acid sequence with at least 95% identity to SEQ ID NO: 52.Attorney Docket No. TP385350WO1

[0111] In some embodiments, the isolated polypeptide comprises two additional CH2 domains N-terminal to the Fc domain connected via a hinge and optionally one or more linkers and comprises “LFLEPS” mutations within the lower hinge and CH2 domains. The “LFLEPS” mutations can comprise the following amino acid substitutions: L234F, L235E and P331S. The isolated polypeptide comprising two additional CH2 domains N-terminal to the Fc domain connected via a hinge and optionally one or more linkers and comprises “LFLEPS” mutations within the lower hinge and CH2 domains comprises SEQ ID NO: 54. The isolated polypeptide comprising two additional CH2 domains N-terminal to the Fc domain connected via a hinge and optionally one or more linkers and comprises “LFLEPS” mutations within the lower hinge and CH2 domains can comprise an amino acid sequence with at least 90% identity to SEQ ID NO: 54. The isolated polypeptide comprising two additional CH2 domains N-terminal to the Fc domain connected via a hinge and optionally one or more linkers and comprises “LFLEPS” mutations within the lower hinge and CH2 domains can comprise an amino acid sequence with at least 95% identity to SEQ ID NO: 54.

[0112] Any isolated polypeptide disclosed herein can be linked to a second polypeptide by one or more disulfide bonds, wherein the isolated polypeptide and the second polypeptide share the same amino acid sequence. The one or more disulfide bonds can occur between a hinge of the isolated polypeptide and a hinge of the second polypeptide.

[0113] Any isolated polypeptide disclosed herein can further comprise a CH3 domain. In certain embodiments, isolated polypeptides disclosed herein further comprises no more than one CH3 domain. In some embodiments the CH3 domain comprises the amino acid sequence of SEQ ID NO: 20. The CH3 domain can comprise an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 20. The CH3 domain can comprise an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 20.

[0114] In some embodiments, the isolated polypeptide comprising a CH3 domain comprises SEQ ID NO: 106. The isolated polypeptide comprising a CH3 domain can comprise an amino acid sequence with at least 90% identity to SEQ ID NO: 106. The isolated polypeptide comprising a CH3 domain can comprise an amino acid sequence with at least 95% identity to SEQ ID NO: 106.

[0115] In some embodiments, the isolated polypeptide comprising a CH3 domain comprises SEQ ID NO: 141. The isolated polypeptide comprising a CH3 domain can comprise an amino acid sequence with at least 90% identity to SEQ ID NO: 141. The isolatedAttorney Docket No. TP385350WO1 polypeptide comprising a CH3 domain can comprise an amino acid sequence with at least 95% identity to SEQ ID NO: 141.

[0116] Also provided herein are antibodies comprising any isolated peptide disclosed herein linked to a Fab domain. The Fab can comprise a CH1 domain. The CH1 domain can comprise the amino acid sequence of SEQ ID NO: 17. The CH1 domain can comprise an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 17. The CH1 domain can comprise an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 17. The Fab can comprise a Fab heavy chain and a Fab light chain that pair to form a binding domain.

[0117] In some embodiments, the Fab can comprise a CH1 domain. The CH1 domain can comprise the amino acid sequence of SEQ ID NO: 100. The CH1 domain can comprise an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 100. The CH1 domain can comprise an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 100. The Fab can comprise a Fab heavy chain and a Fab light chain that pair to form a binding domain.

[0118] In some embodiments, the Fab can comprise a CH1 domain. The CH1 domain can comprise the amino acid sequence of SEQ ID NO: 160. The CH1 domain can comprise an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 160. The CH1 domain can comprise an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 160. The Fab can comprise a Fab heavy chain and a Fab light chain that pair to form a binding domain.

[0119] In some embodiments, the antibodies comprising any isolated peptide disclosed herein linked to a Fab domain have a reduction of background signal of at least 1.5- fold, compared to a corresponding wildtype antibody. In some embodiments, the reduction of background signal is at least 1.5-fold, at least 2-fold, at least 2.5-fold, at least 3-fold, at least 3.5-fold, at least 4-fold, at least 4.5-fold or at least 5-fold compared to a corresponding wild type antibody.

[0120] In some embodiments, the antibodies comprising any isolated peptide disclosed herein linked to a Fab domain have an increase in sensitivity of at least 1.5-fold compared to a corresponding wild type antibody. In some embodiments, the increase in sensitivity is at least 1.5-fold, at least 2-fold, at least 2.5-fold, at least 3-fold, at least 3.5-fold, at least 4-fold, at least 4.5-fold, at least 5-fold, at least 5.5-fold, at least 6-fold, at least 6.5-Attorney Docket No. TP385350WO1 fold, at least 7-fold, at least 7.5-fold, at least 8-fold, at least 8.5-fold, at least 9-fold, at least 9.5-fold, or at least 10-fold compared to a corresponding wild type antibody.

[0121] In some embodiments, the antibodies comprising any isolated peptide disclosed herein linked to a Fab domain has an increase in signal enhancement of at least 1.5- fold compared to a corresponding wild type antibody. In some embodiments, the increase in signal enhancement is at least 1.5-fold, at least 2-fold, at least 2.5-fold, at least 3-fold, at least 3.5-fold, at least 4-fold, at least 4.5-fold, at least 5-fold, at least 5.5-fold, at least 6-fold, at least 6.5-fold, at least 7-fold, at least 7.5-fold, at least 8-fold, at least 8.5-fold, at least 9-fold, at least 9.5-fold, or at least 10-fold compared to a corresponding wild type antibody.

[0122] The Fab of the antibody can immunospecifically bind to one or more target molecules. The Fab can immunospecifically bind to Parkin, OCT4, IDO1, FLAG-tag, pERK1 / 2, PSD-95, or TIGIT, or the Fab can be a rabbit anti-mouse (RAM) IgG secondary antibody.

[0123] In some embodiments, the Fab immunospecifically binds to Parkin. The Fab that immunospecifically binds to Parkin can comprise a Fab heavy chain comprising a heavy chain complementarity determining region 1 (HCDR1) comprising the amino acid sequence of SEQ ID NO: 4, a HCDR2 comprising the amino acid sequence of SEQ ID NO: 5, and a HCDR3 comprising the amino acid sequence of SEQ ID NO: 6. In certain embodiments throughout this disclosure, the CDRs are defined according to the IMGT system.

[0124] The Fab that immunospecifically binds to Parkin can comprise a Fab heavy chain comprising a variable region comprising the amino acid sequence of SEQ ID NO: 7. The Fab that immunospecifically binds to Parkin can comprise a Fab heavy chain comprising a variable region comprising an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 7. The Fab that immunospecifically binds to Parkin can comprise a Fab heavy chain comprising a variable region comprising an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 7.

[0125] The Fab that immunospecifically binds to Parkin can comprise a Fab heavy chain comprising a variable region comprising the amino acid sequence of SEQ ID NO: 9. The Fab that immunospecifically binds to Parkin can comprise a Fab heavy chain comprising a variable region comprising an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 9. The Fab that immunospecifically binds to Parkin can comprise a Fab heavyAttorney Docket No. TP385350WO1 chain comprising a variable region comprising an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 9.

[0126] The Fab that immunospecifically binds to Parkin can comprise a Fab light chain comprising a light chain complementarity determining region 1 (LCDR1) comprising the amino acid sequence of SEQ ID NO: 13, a LCDR2 comprising the amino acid sequence RAS, and a LCDR3 comprising the amino acid sequence of SEQ ID NO: 15.

[0127] The Fab that immunospecifically binds to Parkin can comprise a Fab light chain comprising a variable region comprising the amino acid sequence of SEQ ID NO: 16. The Fab that immunospecifically binds to Parkin can comprise a Fab light chain comprising a variable region comprising an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 16. The Fab that immunospecifically binds to Parkin can comprise a Fab light chain comprising a variable region comprising an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 16.

[0128] The Fab that immunospecifically binds to Parkin can comprise a Fab light chain comprising a variable region comprising the amino acid sequence of SEQ ID NO: 12. The Fab that immunospecifically binds to Parkin can comprise a Fab light chain comprising a variable region comprising an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 12. The Fab that immunospecifically binds to Parkin can comprise a Fab light chain comprising a variable region comprising an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 12.

[0129] The Fab that immunospecifically binds to Parkin can comprise a Fab heavy chain comprising a heavy chain complementarity determining region 1 (HCDR1) comprising the amino acid sequence of SEQ ID NO: 4, a HCDR2 comprising the amino acid sequence of SEQ ID NO: 5, and a HCDR3 comprising the amino acid sequence of SEQ ID NO: 6, and a Fab light chain comprising a light chain complementarity determining region 1 (LCDR1) comprising the amino acid sequence of SEQ ID NO: 13, a LCDR2 comprising the amino acid sequence RAS, and a LCDR3 comprising the amino acid sequence of SEQ ID NO: 15.

[0130] The Fab that immunospecifically binds to Parkin can comprise a Fab heavy chain comprising a variable region comprising the amino acid sequence of SEQ ID NO: 7 and a Fab light chain comprising a variable region comprising the amino acid sequence of SEQ ID NO: 16. The Fab that immunospecifically binds to Parkin can comprise a Fab heavy chain comprising a variable region comprising an amino acid sequence with at least 90%Attorney Docket No. TP385350WO1 sequence identity to SEQ ID NO: 7 and a Fab light chain comprising a variable region comprising an amino acid sequence with at least 90% sequence identity to the amino acid sequence of SEQ ID NO: 16. The Fab that immunospecifically binds to Parkin can comprise a Fab heavy chain comprising a variable region comprising an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 7 and a Fab light chain comprising a variable region comprising an amino acid sequence with at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 16.

[0131] The Fab that immunospecifically binds to Parkin can comprise a Fab heavy chain comprising a variable region comprising the amino acid sequence of SEQ ID NO: 9 and a Fab light chain comprising a variable region comprising the amino acid sequence of SEQ ID NO: 12. The Fab that immunospecifically binds to Parkin can comprise a Fab heavy chain comprising a variable region comprising an amino acid sequence with at least 90% sequence identity to the amino acid sequence of SEQ ID NO: 9 and a Fab light chain comprising a variable region comprising an amino acid sequence with at least 90% sequence identity to the amino acid sequence of SEQ ID NO: 12. The Fab that immunospecifically binds to Parkin can comprise a Fab heavy chain comprising a variable region comprising an amino acid sequence with at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 9 and a Fab light chain comprising a variable region comprising an amino acid sequence with at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 12.

[0132] In some embodiments, the antibody comprising a Fab that immunospecifically binds to Parkin comprises the amino acid sequence of SEQ ID NO: 3. The antibody comprising a Fab that immunospecifically binds to Parkin can comprise an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 3. The antibody comprising a Fab that immunospecifically binds to Parkin can comprise an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 3.

[0133] In some embodiments, the Fab immunospecifically binds to OCT4. The Fab that immunospecifically binds to OCT4 can comprise a heavy chain complementarity determining region 1 (HCDR1) comprising the amino acid sequence of SEQ ID NO: 29, a HCDR2 comprising the amino acid sequence of SEQ ID NO: 30, and a HCDR3 comprising the amino acid sequence of SEQ ID NO: 31.

[0134] The Fab that immunospecifically binds to OCT4 can comprise a Fab heavy chain comprising a variable region comprising the amino acid sequence of SEQ ID NO: 32.Attorney Docket No. TP385350WO1 The Fab that immunospecifically binds to OCT4 can comprise a Fab heavy chain comprising a variable region comprising an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 32. The Fab that immunospecifically binds to OCT4 can comprise a Fab heavy chain comprising a variable region comprising an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 32.

[0135] The Fab that immunospecifically binds to OCT4 can comprise a Fab heavy chain comprising a variable region comprising the amino acid sequence of SEQ ID NO: 34. The Fab that immunospecifically binds to OCT4 can comprise a Fab heavy chain comprising a variable region comprising an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 34. The Fab that immunospecifically binds to OCT4 can comprise a Fab heavy chain comprising a variable region comprising an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 34.

[0136] The Fab that immunospecifically binds OCT4 can comprise a Fab light chain comprising a light chain complementarity determining region 1 (LCDR1) comprising the amino acid sequence of SEQ ID NO: 37, a LCDR2 comprising the amino acid sequence GAS, and a LCDR3 comprising the amino acid sequence of SEQ ID NO: 39.

[0137] The Fab that immunospecifically binds OCT4 can comprise a Fab light chain comprising a variable region comprising the amino acid sequence of SEQ ID NO: 40. The Fab that immunospecifically binds OCT4 can comprise a Fab light chain comprising a variable region comprising an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 40. The Fab that immunospecifically binds OCT4 can comprise a Fab light chain comprising a variable region comprising an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 40.

[0138] The Fab that immunospecifically binds to OCT4 can comprise a Fab light chain comprising a variable region comprising the amino acid sequence of SEQ ID NO: 36. The Fab that immunospecifically binds OCT4 can comprise a Fab light chain comprising a variable region comprising an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 36. The Fab that immunospecifically binds OCT4 can comprise a Fab light chain comprising a variable region comprising an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 36.

[0139] The Fab that immunospecifically binds to OCT4 can comprise a Fab heavy chain comprising a heavy chain complementarity determining region 1 (HCDR1) comprisingAttorney Docket No. TP385350WO1 the amino acid sequence of SEQ ID NO: 29, a HCDR2 comprising the amino acid sequence of SEQ ID NO: 30, and a HCDR3 comprising the amino acid sequence of SEQ ID NO: 31, and a Fab light chain comprising a light chain complementarity determining region 1 (LCDR1) comprising the amino acid sequence of SEQ ID NO: 37, a LCDR2 comprising the amino acid sequence GAS, and a LCDR3 comprising the amino acid sequence of SEQ ID NO: 39.

[0140] The Fab that immunospecifically binds to OCT4 can comprise a Fab heavy chain comprising a variable region comprising the amino acid sequence of SEQ ID NO: 32 and a Fab light chain comprising a variable region comprising the amino acid sequence of SEQ ID NO: 40. The Fab that immunospecifically binds to OCT4 can comprise a Fab heavy chain comprising a variable region comprising an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 32 and a Fab light chain comprising a variable region comprising an amino acid sequence with at least 90% sequence identity to the amino acid sequence of SEQ ID NO: 40. The Fab that immunospecifically binds to OCT4 can comprise a Fab heavy chain comprising a variable region comprising an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 32 and a Fab light chain comprising a variable region comprising an amino acid sequence with at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 40.

[0141] The Fab that immunospecifically binds to OCT4 can comprise a Fab heavy chain comprising a variable region comprising the amino acid sequence of SEQ ID NO: 34 and a Fab light chain comprising a variable region comprising the amino acid sequence of SEQ ID NO: 36. The Fab that immunospecifically binds to OCT4 can comprise a Fab heavy chain comprising a variable region comprising an amino acid sequence with at least 90% sequence identity to the amino acid sequence of SEQ ID NO: 34 and a Fab light chain comprising a variable region comprising an amino acid sequence with at least 90% sequence identity to the amino acid sequence of SEQ ID NO: 36. The Fab that immunospecifically binds to OCT4 can comprise a Fab heavy chain comprising a variable region comprising an amino acid sequence with at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 34 and a Fab light chain comprising a variable region comprising an amino acid sequence with at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 36.

[0142] In some embodiments, the antibody comprising a Fab that immunospecifically binds to OCT4 comprises the amino acid sequence of SEQ ID NO: 28.Attorney Docket No. TP385350WO1 The isolated polypeptide comprising a Fab that immunospecifically binds to OCT4 can comprise an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 28. The isolated polypeptide comprising a Fab that immunospecifically binds to OCT4 can comprise an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 28.

[0143] In some embodiments, the Fab immunospecifically binds to IDO1. The Fab that immunospecifically binds to IDO1 can comprise a heavy chain complementarity determining region 1 (HCDR1) comprising the amino acid sequence of SEQ ID NO: 66, a HCDR2 comprising the amino acid sequence of SEQ ID NO: 67, and a HCDR3 comprising the amino acid sequence of SEQ ID NO: 68.

[0144] The Fab that immunospecifically binds to IDO1 can comprise a Fab heavy chain comprising a variable region comprising the amino acid sequence of SEQ ID NO: 65. The Fab that immunospecifically binds to IDO1 can comprise a Fab heavy chain comprising a variable region comprising an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 65. The Fab that immunospecifically binds to IDO1 can comprise a Fab heavy chain comprising a variable region comprising an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 65.

[0145] The Fab that immunospecifically binds IDO1 can comprise a Fab light chain comprising a light chain complementarity determining region 1 (LCDR1) comprising the amino acid sequence of SEQ ID NO: 72, a LCDR2 comprising the amino acid sequence of YAS, and a LCDR3 comprising the amino acid sequence of SEQ ID NO: 73.

[0146] The Fab that immunospecifically binds IDO1 can comprise a Fab light chain comprising a variable region comprising the amino acid sequence of SEQ ID NO: 71. The Fab that immunospecifically binds IDO1 can comprise a Fab light chain comprising a variable region comprising an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 71. The Fab that immunospecifically binds IDO1 can comprise a Fab light chain comprising a variable domain comprising an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 71.

[0147] The Fab that immunospecifically binds to IDO1 can comprise a Fab heavy chain comprising a heavy chain complementarity determining region 1 (HCDR1) comprising the amino acid sequence of SEQ ID NO: 66, a HCDR2 comprising the amino acid sequence of SEQ ID NO: 67, and a HCDR3 comprising the amino acid sequence of SEQ ID NO: 68, and a Fab light chain comprising a light chain complementarity determining region 1Attorney Docket No. TP385350WO1 (LCDR1) comprising the amino acid sequence of SEQ ID NO: 72, a LCDR2 comprising the amino acid sequence of YAS, and a LCDR3 comprising the amino acid sequence of SEQ ID NO: 73.

[0148] The Fab that immunospecifically binds to IDO1 can comprise a Fab heavy chain comprising a variable region comprising the amino acid sequence of SEQ ID NO: 65 and a Fab light chain comprising a variable region comprising the amino acid sequence of SEQ ID NO: 71. The Fab that immunospecifically binds to IDO1 can comprise a Fab heavy chain comprising a variable region comprising an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 65 and a Fab light chain comprising a variable region comprising an amino acid sequence with at least 90% sequence identity to the amino acid sequence of SEQ ID NO: 71. The Fab that immunospecifically binds to IDO1 can comprise a Fab heavy chain comprising a variable region comprising an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 65 and a Fab light chain comprising a variable region comprising an amino acid sequence with at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 71.

[0149] In some embodiments, the antibody comprising a Fab that immunospecifically binds to IDO1 comprises the amino acid sequence of SEQ ID NO: 79. The isolated polypeptide comprising a Fab that immunospecifically binds to IDO1 can comprise an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 79. The isolated polypeptide comprising a Fab that immunospecifically binds to IDO1 can comprise an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 79.

[0150] In some embodiments, the Fab immunospecifically binds to FLAG-tag. The Fab that immunospecifically binds to FLAG-tag can comprise a heavy chain complementarity determining region 1 (HCDR1) comprising the amino acid sequence of SEQ ID NO: 163, a HCDR2 comprising the amino acid sequence of SEQ ID NO: 164, and a HCDR3 comprising the amino acid sequence of SEQ ID NO: 165.

[0151] The Fab that immunospecifically binds to FLAG-tag can comprise a Fab heavy chain comprising a variable region comprising the amino acid sequence of SEQ ID NO: 162. The Fab that immunospecifically binds to FLAG-tag can comprise a Fab heavy chain comprising a variable region comprising an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 162. The Fab that immunospecifically binds to FLAG-tagAttorney Docket No. TP385350WO1 can comprise a Fab heavy chain comprising a variable region comprising an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 162.

[0152] The Fab that immunospecifically binds FLAG-tag can comprise a Fab light chain comprising a light chain complementarity determining region 1 (LCDR1) comprising the amino acid sequence of SEQ ID NO: 168, a LCDR2 comprising the amino acid sequence of YAS, and a LCDR3 comprising the amino acid sequence of SEQ ID NO: 169.

[0153] The Fab that immunospecifically binds FLAG-tag can comprise a Fab light chain comprising a variable region comprising the amino acid sequence of SEQ ID NO: 167. The Fab that immunospecifically binds FLAG-tag can comprise a Fab light chain comprising a variable region comprising an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 167. The Fab that immunospecifically binds FLAG-tag can comprise a Fab light chain comprising a variable domain comprising an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 167.

[0154] The Fab that immunospecifically binds to FLAG-tag can comprise a Fab heavy chain comprising a heavy chain complementarity determining region 1 (HCDR1) comprising the amino acid sequence of SEQ ID NO: 163, a HCDR2 comprising the amino acid sequence of SEQ ID NO: 164, and a HCDR3 comprising the amino acid sequence of SEQ ID NO: 165, and a Fab light chain comprising a light chain complementarity determining region 1 (LCDR1) comprising the amino acid sequence of SEQ ID NO: 168, a LCDR2 comprising the amino acid sequence of YAS, and a LCDR3 comprising the amino acid sequence of SEQ ID NO: 169.

[0155] The Fab that immunospecifically binds to FLAG-tag can comprise a Fab heavy chain comprising a variable region comprising the amino acid sequence of SEQ ID NO: 162 and a Fab light chain comprising a variable region comprising the amino acid sequence of SEQ ID NO: 167. The Fab that immunospecifically binds to FLAG-tag can comprise a Fab heavy chain comprising a variable region comprising an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 162 and a Fab light chain comprising a variable region comprising an amino acid sequence with at least 90% sequence identity to the amino acid sequence of SEQ ID NO: 167. The Fab that immunospecifically binds to FLAG- tag can comprise a Fab heavy chain comprising a variable region comprising an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 162 and a Fab light chainAttorney Docket No. TP385350WO1 comprising a variable region comprising an amino acid sequence with at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 167.

[0156] In some embodiments, the antibody comprising a Fab that immunospecifically binds to FLAG-tag comprises the amino acid sequence of SEQ ID NO: 175. The isolated polypeptide comprising a Fab that immunospecifically binds to FLAG-tag can comprise an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 175. The isolated polypeptide comprising a Fab that immunospecifically binds to FLAG-tag can comprise an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 175.

[0157] In some embodiments, the Fab is a rabbit anti-mouse IgG secondary antibody (also referred to herein as “rabbit anti-mouse IgG antibody,” “rabbit anti-mouse IgG,” or “RAM”). The Fab that is a RAM can comprise a heavy chain complementarity determining region 1 (HCDR1) comprising the amino acid sequence of SEQ ID NO: 127, a HCDR2 comprising the amino acid sequence of SEQ ID NO: 128, and a HCDR3 comprising the amino acid sequence of SEQ ID NO: 129.

[0158] The Fab that is a RAM can comprise a Fab heavy chain comprising a variable region comprising the amino acid sequence of SEQ ID NO: 126. The Fab that is a RAM can comprise a Fab heavy chain comprising a variable region comprising an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 126. The Fab that is a RAM can comprise a Fab heavy chain comprising a variable region comprising an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 126.

[0159] The Fab that is a RAM can comprise a Fab light chain comprising a light chain complementarity determining region 1 (LCDR1) comprising the amino acid sequence of SEQ ID NO: 132, a LCDR2 comprising the amino acid sequence of GAS, and a LCDR3 comprising the amino acid sequence of SEQ ID NO: 133.

[0160] The Fab that is a RAM can comprise a Fab light chain comprising a variable region comprising the amino acid sequence of SEQ ID NO: 131. The Fab that is a RAM can comprise a Fab light chain comprising a variable region comprising an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 131. The Fab that is a RAM can comprise a Fab light chain comprising a variable region comprising an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 131.Attorney Docket No. TP385350WO1

[0161] The Fab that is a RAM can comprise a Fab heavy chain comprising a heavy chain complementarity determining region 1 (HCDR1) comprising the amino acid sequence of SEQ ID NO: 127, a HCDR2 comprising the amino acid sequence of SEQ ID NO: 128, and a HCDR3 comprising the amino acid sequence of SEQ ID NO: 129, and a Fab light chain comprising a light chain complementarity determining region 1 (LCDR1) comprising the amino acid sequence of SEQ ID NO: 132, a LCDR2 comprising the amino acid sequence of GAS, and a LCDR3 comprising the amino acid sequence of SEQ ID NO: 133.

[0162] The Fab that is a RAM can comprise a Fab heavy chain comprising a variable region comprising the amino acid sequence of SEQ ID NO: 126 and a Fab light chain comprising a variable region comprising the amino acid sequence of SEQ ID NO: 131. The Fab that is a RAM can comprise a Fab heavy chain comprising a variable region comprising an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 126 and a Fab light chain comprising a variable region comprising an amino acid sequence with at least 90% sequence identity to the amino acid sequence of SEQ ID NO: 131. The Fab that is a RAM can comprise a Fab heavy chain comprising a variable region comprising an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 126 and a Fab light chain comprising a variable region comprising an amino acid sequence with at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 131.

[0163] In some embodiments, the antibody comprising a Fab that is a RAM comprises the amino acid sequence of SEQ ID NO: 139. The isolated polypeptide comprising a RAM can comprise an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 139. The isolated polypeptide comprising a RAM can comprise an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 139.

[0164] In some embodiments, the Fab immunospecifically binds to pERK1 / 2. The Fab that immunospecifically binds to pERK1 / 2 can comprise a heavy chain complementarity determining region 1 (HCDR1) comprising the amino acid sequence of SEQ ID NO: 85, a HCDR2 comprising the amino acid sequence of SEQ ID NO: 86, and a HCDR3 comprising the amino acid sequence of SEQ ID NO: 87.

[0165] The Fab that immunospecifically binds to pERK1 / 2 can comprise a Fab heavy chain comprising a variable region comprising the amino acid sequence of SEQ ID NO: 84. The Fab that immunospecifically binds to pERK1 / 2 can comprise a Fab heavy chain comprising a variable region comprising an amino acid sequence with at least 90% sequenceAttorney Docket No. TP385350WO1 identity to SEQ ID NO: 84. The Fab that immunospecifically binds to pERK1 / 2 can comprise a Fab heavy chain comprising a variable region comprising an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 84.

[0166] The Fab that immunospecifically binds pERK1 / 2 can comprise a Fab light chain comprising a light chain complementarity determining region 1 (LCDR1) comprising the amino acid sequence of SEQ ID NO: 91, a LCDR2 comprising the amino acid sequence of KVS, and a LCDR3 comprising the amino acid sequence of SEQ ID NO: 92.

[0167] The Fab that immunospecifically binds pERK1 / 2 can comprise a Fab light chain comprising a variable region comprising the amino acid sequence of SEQ ID NO: 90. The Fab that immunospecifically binds pERK1 / 2 can comprise a Fab light chain comprising a variable region comprising an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 90. The Fab that immunospecifically binds pERK1 / 2 can comprise a Fab light chain comprising a variable region comprising an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 90.

[0168] The Fab that immunospecifically binds to pERK1 / 2 can comprise a Fab heavy chain comprising a heavy chain complementarity determining region 1 (HCDR1) comprising the amino acid sequence of SEQ ID NO: 85, a HCDR2 comprising the amino acid sequence of SEQ ID NO: 86, and a HCDR3 comprising the amino acid sequence of SEQ ID NO: 87, and a Fab light chain comprising a light chain complementarity determining region 1 (LCDR1) comprising the amino acid sequence of SEQ ID NO: 91, a LCDR2 comprising the amino acid sequence of KVS, and a LCDR3 comprising the amino acid sequence of SEQ ID NO: 92.

[0169] The Fab that immunospecifically binds to pERK1 / 2 can comprise a Fab heavy chain comprising a variable region comprising the amino acid sequence of SEQ ID NO: 84 and a Fab light chain comprising a variable region comprising the amino acid sequence of SEQ ID NO: 90. The Fab that immunospecifically binds to pERK1 / 2 can comprise a Fab heavy chain comprising a variable region comprising an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 84 and a Fab light chain comprising a variable region comprising an amino acid sequence with at least 90% sequence identity to the amino acid sequence of SEQ ID NO: 90. The Fab that immunospecifically binds to pERK1 / 2 can comprise a Fab heavy chain comprising a variable region comprising an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 84 and a Fab light chainAttorney Docket No. TP385350WO1 comprising a variable region comprising an amino acid sequence with at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 90.

[0170] In some embodiments, the antibody comprising a Fab that immunospecifically binds to pERK1 / 2 comprises the amino acid sequence of SEQ ID NO: 98. The isolated polypeptide comprising a Fab that immunospecifically binds to pERK1 / 2 can comprise an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 98. The isolated polypeptide comprising a Fab that immunospecifically binds to pERK1 / 2 can comprise an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 98.

[0171] In some embodiments, the Fab immunospecifically binds to PSD-95. The Fab that immunospecifically binds to PSD-95 can comprise a heavy chain complementarity determining region 1 (HCDR1) comprising the amino acid sequence of SEQ ID NO: 111, a HCDR2 comprising the amino acid sequence of SEQ ID NO: 112, and a HCDR3 comprising the amino acid sequence of SEQ ID NO: 113.

[0172] The Fab that immunospecifically binds to PSD-95 can comprise a Fab heavy chain comprising a variable region comprising the amino acid sequence of SEQ ID NO: 110. The Fab that immunospecifically binds to PSD-95 can comprise a Fab heavy chain comprising a variable region comprising an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 110. The Fab that immunospecifically binds to PSD-95 can comprise a Fab heavy chain comprising a variable region comprising an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 110.

[0173] The Fab that immunospecifically binds PSD-95 can comprise a Fab light chain comprising a light chain complementarity determining region 1 (LCDR1) comprising the amino acid sequence of SEQ ID NO: 116, a LCDR2 comprising the amino acid sequence of DTS, and a LCDR3 comprising the amino acid sequence of SEQ ID NO: 117.

[0174] The Fab that immunospecifically binds PSD-95 can comprise a Fab light chain comprising a variable region comprising the amino acid sequence of SEQ ID NO: 115. The Fab that immunospecifically binds PSD-95 can comprise a Fab light chain comprising a variable region comprising an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 115. The Fab that immunospecifically binds PSD-95 can comprise a Fab light chain comprising a variable region comprising an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 115.Attorney Docket No. TP385350WO1

[0175] The Fab that immunospecifically binds to PSD-95 can comprise a Fab heavy chain comprising a heavy chain complementarity determining region 1 (HCDR1) comprising the amino acid sequence of SEQ ID NO: 111, a HCDR2 comprising the amino acid sequence of SEQ ID NO: 112, and a HCDR3 comprising the amino acid sequence of SEQ ID NO: 113, and a Fab light chain comprising a light chain complementarity determining region 1 (LCDR1) comprising the amino acid sequence of SEQ ID NO: 116, a LCDR2 comprising the amino acid sequence of DTS, and a LCDR3 comprising the amino acid sequence of SEQ ID NO: 117.

[0176] The Fab that immunospecifically binds to PSD-95 can comprise a Fab heavy chain comprising a variable region comprising the amino acid sequence of SEQ ID NO: 110 and a Fab light chain comprising a variable region comprising the amino acid sequence of SEQ ID NO: 115. The Fab that immunospecifically binds to PSD-95 can comprise a Fab heavy chain comprising a variable region comprising an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 110 and a Fab light chain comprising a variable region comprising an amino acid sequence with at least 90% sequence identity to the amino acid sequence of SEQ ID NO: 115. The Fab that immunospecifically binds to PSD-95 can comprise a Fab heavy chain comprising a variable region comprising an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 110 and a Fab light chain comprising a variable region comprising an amino acid sequence with at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 115.

[0177] In some embodiments, the antibody comprising a Fab that immunospecifically binds to PSD-95 comprises the amino acid sequence of SEQ ID NO: 123. The isolated polypeptide comprising a Fab that immunospecifically binds to PSD-95 can comprise an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 123. The isolated polypeptide comprising a Fab that immunospecifically binds to PSD-95 can comprise an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 123.

[0178] In some embodiments, the Fab immunospecifically binds to TIGIT. The Fab that immunospecifically binds to TIGIT can comprise a heavy chain complementarity determining region 1 (HCDR1) comprising the amino acid sequence of SEQ ID NO: 146, a HCDR2 comprising the amino acid sequence of SEQ ID NO: 147, and a HCDR3 comprising the amino acid sequence of SEQ ID NO: 148.Attorney Docket No. TP385350WO1

[0179] The Fab that immunospecifically binds to TIGIT can comprise a Fab heavy chain comprising a variable region comprising the amino acid sequence of SEQ ID NO: 145. The Fab that immunospecifically binds to TIGIT can comprise a Fab heavy chain comprising a variable region comprising an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 145. The Fab that immunospecifically binds to TIGIT can comprise a Fab heavy chain comprising a variable region comprising an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 145.

[0180] The Fab that immunospecifically binds to TIGIT can comprise a Fab light chain comprising a light chain complementarity determining region 1 (LCDR1) comprising the amino acid sequence of SEQ ID NO: 151, a LCDR2 comprising the amino acid sequence of SAS, and a LCDR3 comprising the amino acid sequence of SEQ ID NO: 152.

[0181] The Fab that immunospecifically binds to TIGIT can comprise a Fab light chain comprising a variable region comprising the amino acid sequence of SEQ ID NO: 150. The Fab that immunospecifically binds to TIGIT can comprise a Fab light chain comprising a variable region comprising an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 150. The Fab that immunospecifically binds to TIGIT can comprise a Fab light chain comprising a variable region comprising an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 150.

[0182] The Fab that immunospecifically binds to TIGIT can comprise a Fab heavy chain comprising a heavy chain complementarity determining region 1 (HCDR1) comprising the amino acid sequence of SEQ ID NO: 146, a HCDR2 comprising the amino acid sequence of SEQ ID NO: 147, and a HCDR3 comprising the amino acid sequence of SEQ ID NO: 148, and a Fab light chain comprising a light chain complementarity determining region 1 (LCDR1) comprising the amino acid sequence of SEQ ID NO: 151, a LCDR2 comprising the amino acid sequence of SAS, and a LCDR3 comprising the amino acid sequence of SEQ ID NO: 152.

[0183] The Fab that immunospecifically binds to TIGIT can comprise a Fab heavy chain comprising a variable region comprising the amino acid sequence of SEQ ID NO: 145 and a Fab light chain comprising a variable region comprising the amino acid sequence of SEQ ID NO: 150. The Fab that immunospecifically binds to TIGIT can comprise a Fab heavy chain comprising a variable region comprising an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 145 and a Fab light chain comprising a variable regionAttorney Docket No. TP385350WO1 comprising an amino acid sequence with at least 90% sequence identity to the amino acid sequence of SEQ ID NO: 150. The Fab that immunospecifically binds to TIGIT can comprise a Fab heavy chain comprising a variable region comprising an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 145 and a Fab light chain comprising a variable region comprising an amino acid sequence with at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 150.

[0184] In some embodiments, the Fab that immunospecifically binds to TIGIT comprises the amino acid sequence of SEQ ID NO: 157. The isolated polypeptide comprising a Fab that immunospecifically binds to TIGIT can comprise an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 157. The isolated polypeptide comprising a Fab that immunospecifically binds to TIGIT can comprise an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 157.

[0185] Any antibody disclosed herein can be an antibody with the isotype IgG.

[0186] In some embodiments, the isolated polypeptide or the antibody further comprises a label.

[0187] Disclosed herein are nucleic acid molecules comprising, consisting essentially of, or consisting of polynucleotides encoding isolated polypeptides comprising, consisting essentially of, or consisting of the amino acid sequence of Formula I: Z-(X-Yn- Zr)p-X, wherein Z is a hinge, X is a CH2 domain, Y is a linker, n is an integer of 1 or greater, p is an integer of 2 or greater, and r is an integer of 0 or 1.

[0188] The nucleic acid molecules can comprise polynucleotides that encode a hinge (Z) comprising the amino acid sequence of SEQ ID NO: 18. The polynucleotides can encode a hinge (Z) comprising an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 18. The polynucleotides can encode a hinge (Z) comprising an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 18. In some embodiments, the polynucleotides encoding the hinge (Z) comprise SEQ ID NO: 22.

[0189] The nucleic acid molecules can comprise polynucleotides that encode a polypeptide that comprises, consists essentially of, or consists of (X -Yn-Zr)p, wherein the hinge (Z) comprises the amino acid sequence of SEQ ID NO: 48. The polynucleotides can encode a hinge (Z) comprising an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 48. The polynucleotides can encode a hinge (Z) comprising an amino acidAttorney Docket No. TP385350WO1 sequence with at least 95% sequence identity to SEQ ID NO: 48. In some embodiments, the polynucleotides encoding the hinge (Z) comprise SEQ ID NO: 51.

[0190] The nucleic acid molecules can comprise polynucleotides that encode a polypeptide that comprises, consists essentially of, or consists of (X-Yn-Zr), wherein the hinge (Z) comprises the amino acid sequence of SEQ ID NO: 102. The polynucleotides can encode a hinge (Z) comprising an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 102. The polynucleotides can encode a hinge (Z) comprising an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 102. In some embodiments, the polynucleotides encoding the hinge (Z) comprise SEQ ID NO: 101.

[0191] The polynucleotides encoding a polypeptide comprising, consisting essentially of, or consisting of (X-Yn-Zr)p can encode a CH2 domain (X) comprising the amino acid sequence of SEQ ID NO: 19. The polynucleotides can encode a CH2 domain (X) comprising an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 19. The polynucleotides can encode a CH2 domain (X) comprising an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 19. In some embodiments, the polynucleotides encoding the CH2 domain (X) comprise SEQ ID NO: 23 or SEQ ID NO: 178.

[0192] The nucleic acid molecules encoding a polypeptide comprising, consisting essentially of, or consisting of (X-Yn-Zr)p, can comprise polynucleotides that encode a CH2 domain (X) comprising the amino acid sequence of SEQ ID NO: 49. The polynucleotides can encode a CH2 domain (X) comprising an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 49. The polynucleotides can encode a CH2 domain (X) comprising an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 49. In some embodiments, the polynucleotides encoding the CH2 domain (X) comprise SEQ ID NO: 62.

[0193] The polynucleotides encoding a polypeptide comprising, consisting essentially of, or consisting of (X-Yn-Zr)p can encode a CH2 domain (X) comprising the amino acid sequence of SEQ ID NO: 102. The polynucleotides can encode a CH2 domain (X) comprising an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 102. The polynucleotides can encode a CH2 domain (X) comprising an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 102. In some embodiments, the polynucleotides encoding the CH2 domain (X) comprise SEQ ID NO: 101.

[0194] The nucleic acid molecules encoding a polypeptide comprising, consisting essentially of, or consisting of (X-Yn-Zr)p can comprise polynucleotides that encode anAttorney Docket No. TP385350WO1 isolated polypeptide comprising one or more linkers (Y) comprising the amino acid sequence of GS, GGS, SEQ ID NO: 14, SEQ ID NO: 38, SEQ ID NO: 43, or SEQ ID NO: 44. The polynucleotide can encode one or more linkers (Y) comprising GS. The polynucleotide can encode one or more linkers (Y) comprising GGS. The polynucleotide can encode one or more linkers (Y) comprising SEQ ID NO: 14. The polynucleotide can encode one or more linkers (Y) comprising SEQ ID NO: 38. The polynucleotide can encode one or more linkers (Y) comprising SEQ ID NO: 43. The polynucleotide can encode one or more linkers (Y) comprising an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 43. The polynucleotide can encode one or more linkers (Y) comprising SEQ ID NO: 44. The polynucleotide can encode one or more linkers (Y) comprising an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 44. In some embodiments, the polynucleotide encoding the one or more linkers (Y) comprise SEQ ID NO: 46.

[0195] The polynucleotide can encode an isolated polypeptide comprising one linker (Y), such that n is 1. The polynucleotide can encode an isolated polypeptide comprising two linkers (Y), such that n is 2. The polynucleotide can encode an isolated polypeptide comprising three linkers (Y), such that n is 3. The polynucleotide can encode an isolated polypeptide comprising four linkers (Y), such that n is 4. The polynucleotide can encode an isolated polypeptide comprising five linkers (Y), such that n is 5.

[0196] The polynucleotide can encode an isolated polypeptide comprising two or more regions comprising a CH2-linker region ((X-Yn)p), such that p is 2 or greater. The polynucleotide can encode an isolated polypeptide comprising two CH2-linker regions ((X- Yn)p), such that p is 2. The polynucleotide can encode an isolated polypeptide comprising three CH2-linker regions ((X-Yn)p), such that p is 3. The polynucleotide can encode an isolated polypeptide comprising four CH2-linker regions ((X-Yn)p), such that p is 4. The polynucleotide can encode an isolated polypeptide comprising five CH2-linker regions ((X- Yn)p), such that p is 5.

[0197] The polynucleotide can encode an isolated polypeptide comprising two or more regions comprising a CH2-linker-hinge region ((X-Yn-Zr)p), such that p is 2 or greater. The polynucleotide can encode an isolated polypeptide comprising two CH2-linker-hinge regions ((X-Yn-Zr)p), such that p is 2. The polynucleotide can encode an isolated polypeptide comprising three CH2-linker-hinge regions ((X-Yn-Zr)p), such that p is 3. The polynucleotide can encode an isolated polypeptide comprising four CH2-linker-hinge regions ((X-Yn-Zr)p),Attorney Docket No. TP385350WO1 such that p is 4. The polynucleotide can encode an isolated polypeptide comprising five CH2- linker-hinge regions ((X-Yn-Zr)p), such that p is 5.

[0198] In some embodiments, the nucleic acid molecule comprises a polynucleotide encoding an isolated polypeptide comprising, consisting essentially of, or consisting of: Z (hinge) comprising the amino acid sequence of SEQ ID NO: 18, X (CH2 domain) comprising the amino acid sequence of SEQ ID NO: 19, Y (linker) comprising the amino acid sequence of SEQ ID NO: 44, wherein n is 1, and p is 2. In some embodiments, the polynucleotide encoding the hinge (X) comprises SEQ ID NO: 22 and the polynucleotide encoding the CH2 domain (X) comprises SEQ ID NO: 23 or SEQ ID NO: 178.

[0199] In some embodiments, the nucleic acid molecule comprises a polynucleotide encoding an isolated polypeptide comprising, consisting essentially of, or consisting of: Z (hinge) comprising the amino acid sequence of SEQ ID NO: 48, X (CH2 domain) comprising the amino acid sequence of SEQ ID NO: 49, Y (linker) comprising the amino acid sequence of SEQ ID NO: 44, wherein n is 1, and p is 2. In some embodiments, the polynucleotide encoding the hinge (X) comprises SEQ ID NO: 51 and the polynucleotide encoding the CH2 domain (X) comprises SEQ ID NO: 62.

[0200] In some embodiments, the nucleic acid molecule comprises a polynucleotide encoding an isolated polypeptide comprising, consisting essentially of, or consisting of: Z (hinge) comprising the amino acid sequence of SEQ ID NO: 102, X (CH2 domain) comprising the amino acid sequence of SEQ ID NO: 104, Y (linker) comprising the amino acid sequence of SEQ ID NO: 44 wherein n is 1, r is 0 or 1, and p is 2. In some embodiments, the polynucleotide encoding the hinge (X) comprises SEQ ID NO: 101 and the polynucleotide encoding the CH2 domain (X) comprises SEQ ID NO: 103.

[0201] The polynucleotide can encode an isolated polypeptide comprising two additional CH2 domains N-terminal to the Fc domain connected via a hinge and optionally one or more linkers, wherein the isolated polypeptide comprises the amino acid sequence of SEQ ID NO: 52. In some embodiments, the polynucleotide encodes an isolated polypeptide comprising two additional CH2 domains N-terminal to the Fc domain connected via a hinge and optionally one or more linkers, the isolated polypeptide comprising SEQ ID NO: 52. The polynucleotide can encode an isolated polypeptide comprising two additional CH2 domains N-terminal to the Fc domain connected via a hinge and optionally one or more linkers, the isolated polypeptide comprising an amino acid sequence with at least 90% identity to SEQ IDAttorney Docket No. TP385350WO1 NO: 52. The polynucleotide can encode an isolated polypeptide comprising two additional CH2 domains N-terminal to the Fc domain connected via a hinge and optionally one or more linkers, the isolated polypeptide comprising an amino acid sequence with at least 95% identity to SEQ ID NO: 52. In some embodiments, in the polynucleotide encoding an isolated polypeptide comprising two additional CH2 domains N-terminal to the Fc domain connected via a hinge and optionally one or more linkers comprises SEQ ID NO: 53.

[0202] In some embodiments, the polynucleotide encodes an isolated polypeptide comprising two additional CH2 domains N-terminal to the Fc domain connected via a hinge and optionally one or more linkers and comprises “LFLEPS” mutations within the lower hinge and CH2 domains. In some embodiments, the “LFLEPS” mutations can comprise the following amino acid substitutions: L234F, L235E and P331S. In some embodiments, the polynucleotide encodes an isolated polypeptide comprising two additional CH2 domains N- terminal to the Fc domain connected via a hinge and optionally one or more linkers and comprises “LFLEPS” mutations within the lower hinge and CH2 domains, the isolated polypeptide comprising SEQ ID NO: 54. The polynucleotide can encode an isolated polypeptide comprising two additional CH2 domains N-terminal to the Fc domain connected via a hinge and optionally one or more linkers and comprises “LFLEPS” mutations within the lower hinge and CH2 domains, the isolated polypeptide comprising an amino acid sequence with at least 90% identity to SEQ ID NO: 54. The polynucleotide can encode an isolated polypeptide comprising two additional CH2 domains N-terminal to the Fc domain connected via a hinge and optionally one or more linkers and comprises “LFLEPS” mutations within the lower hinge and CH2 domains, the isolated polypeptide comprising an amino acid sequence with at least 95% identity to SEQ ID NO: 54. In some embodiments, in the polynucleotide encoding an isolated polypeptide comprising two additional CH2 domains N-terminal to the Fc domain connected via a hinge and optionally one or more linkers and comprises “LFLEPS” mutations within the lower hinge and CH2 domains comprises SEQ ID NO: 55.

[0203] The polynucleotide can encode any isolated polypeptide disclosed herein and can further encode a second polypeptide, wherein the isolated polypeptide and the second polypeptide share the same amino acid sequence.

[0204] The polynucleotide can encode any isolated polypeptide disclosed herein that further comprises a CH3 domain. In some embodiments the polynucleotide encodes a CH3 domain comprising the amino acid sequence of SEQ ID NO: 20. The polynucleotide canAttorney Docket No. TP385350WO1 encode a CH3 domain comprising an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 20. The polynucleotide can encode a CH3 domain comprising an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 20. In some embodiments, the polynucleotide encoding the CH3 domain comprises SEQ ID NO: 24 or SEQ ID NO: 179.

[0205] In some embodiments, the polynucleotide encodes an isolated polypeptide comprising a CH3 domain, the isolated polypeptide comprising SEQ ID NO: 106. The polynucleotide can encode an isolated polypeptide comprising a CH3 domain, the isolated polypeptide comprising an amino acid sequence with at least 90% identity to SEQ ID NO: 106. The polynucleotide can encode an isolated polypeptide comprising a CH3 domain, the isolated polypeptide comprising an amino acid sequence with at least 95% identity to SEQ ID NO: 106. In some embodiments, in the polynucleotide encoding an isolated polypeptide comprising a CH3 domain comprises SEQ ID NO: 105.

[0206] In some embodiments, the polynucleotide encodes an isolated polypeptide comprising a CH3 domain, the isolated polypeptide comprising SEQ ID NO: 141. The polynucleotide can encode an isolated polypeptide comprising a CH3 domain, the isolated polypeptide comprising an amino acid sequence with at least 90% identity to SEQ ID NO: 141. The polynucleotide can encode an isolated polypeptide comprising a CH3 domain, the isolated polypeptide comprising an amino acid sequence with at least 95% identity to SEQ ID NO: 141. In some embodiments, in the polynucleotide encoding an isolated polypeptide comprising a CH3 domain comprises SEQ ID NO: 140.

[0207] The polynucleotide can encode any antibody comprising any isolated polypeptide disclosed herein that further comprises a Fab. The polynucleotide can encode a Fab comprising a CH1 domain. The polynucleotide can encode a CH1 domain comprising the amino acid sequence of SEQ ID NO: 17. The polynucleotide can encode a CH1 domain comprising an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 17. The polynucleotide can encode a CH1 domain comprising amino acid sequence with at least 95% sequence identity to SEQ ID NO: 17. The polynucleotide encoding a CH1 domain can comprise SEQ ID NO: 21, SEQ ID NO: 176, or SEQ ID NO: 177. The polynucleotide can encode a Fab comprising a Fab heavy chain and a Fab light chain that pair to form a binding domain.Attorney Docket No. TP385350WO1

[0208] The polynucleotide can encode any antibody comprising any isolated polypeptide disclosed herein that further comprises a Fab. The polynucleotide can encode a Fab comprising a CH1 domain. The polynucleotide can encode a CH1 domain comprising the amino acid sequence of SEQ ID NO: 100. The polynucleotide can encode a CH1 domain comprising an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 100. The polynucleotide can encode a CH1 domain comprising amino acid sequence with at least 95% sequence identity to SEQ ID NO: 100. The polynucleotide encoding a CH1 domain can comprise SEQ ID NO: 99. The polynucleotide can encode a Fab comprising a Fab heavy chain and a Fab light chain that pair to form a binding domain.

[0209] The polynucleotide can encode any antibody comprising any isolated polypeptide disclosed herein that further comprises a Fab. The polynucleotide can encode a Fab comprising a CH1 domain. The polynucleotide can encode a CH1 domain comprising the amino acid sequence of SEQ ID NO: 160. The polynucleotide can encode a CH1 domain comprising an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 160. The polynucleotide can encode a CH1 domain comprising amino acid sequence with at least 95% sequence identity to SEQ ID NO: 160. The polynucleotide encoding a CH1 domain can comprise SEQ ID NO: 159. The polynucleotide can encode a Fab comprising a Fab heavy chain and a Fab light chain that pair to form a binding domain.

[0210] The polynucleotide can encode a Fab that immunospecifically binds to one or more target molecules. The polynucleotide can encode a Fab that immunospecifically binds to Parkin, OCT4, IDO1, FLAG-tag, pERK1 / 2, PSD-95, or TIGIT, or the Fab can be a rabbit anti-mouse (RAM) IgG secondary antibody.

[0211] In some embodiments, the polynucleotide can encode a Fab that immunospecifically binds to Parkin. The polynucleotide can encode a Fab that immunospecifically binds to Parkin, the Fab comprising a Fab heavy chain comprising a heavy chain complementarity determining region 1 (HCDR1) comprising the amino acid sequence of SEQ ID NO: 4, a HCDR2 comprising the amino acid sequence of SEQ ID NO: 5, and a HCDR3 comprising the amino acid sequence of SEQ ID NO: 6.

[0212] The polynucleotide can encode a Fab that immunospecifically binds to Parkin, the Fab comprising a Fab heavy chain comprising a variable region comprising the amino acid sequence of SEQ ID NO: 7. The polynucleotide can encode a Fab heavy chain comprising a variable region comprising an amino acid sequence with at least 90% sequenceAttorney Docket No. TP385350WO1 identity to SEQ ID NO: 7. The polynucleotide can encode a Fab heavy chain comprising a variable region comprising an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 7.

[0213] The polynucleotide can encode a Fab that immunospecifically binds to Parkin, the Fab comprising a Fab heavy chain comprising a variable region comprising the amino acid sequence of SEQ ID NO: 9. The polynucleotide can encode a Fab heavy chain comprising a variable region comprising an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 9. The polynucleotide can encode a Fab heavy chain comprising a variable region comprising an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 9. In some embodiments, in the polynucleotide encoding a Fab that immunospecifically binds to Parkin, the polynucleotide comprises SEQ ID NO: 8.

[0214] The polynucleotide can encode a Fab that immunospecifically binds to Parkin, the Fab comprising a Fab light chain comprising a light chain complementarity determining region 1 (LCDR1) comprising the amino acid sequence of SEQ ID NO: 13, a LCDR2 comprising the amino acid sequence RAS, and a LCDR3 comprising the amino acid sequence of SEQ ID NO: 15.

[0215] The polynucleotide can encode a Fab that immunospecifically binds to Parkin, the Fab comprising a Fab light chain comprising a variable region comprising the amino acid sequence of SEQ ID NO: 16. The polynucleotide can encode a Fab light chain comprising a variable region comprising an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 16. The polynucleotide can encode a Fab light chain comprising a variable region comprising an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 16.

[0216] The polynucleotide can encode a Fab that immunospecifically binds to Parkin, the Fab comprising a Fab light chain comprising a variable region comprising the amino acid sequence of SEQ ID NO: 12. The polynucleotide can encode a Fab light chain comprising a variable region comprising an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 12. The polynucleotide can encode a Fab light chain comprising a variable region comprising an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 12. In some embodiments, in the polynucleotide encoding a Fab that immunospecifically binds to Parkin, the polynucleotide comprises SEQ ID NO: 11.Attorney Docket No. TP385350WO1

[0217] The polynucleotide can encode a Fab that immunospecifically binds to Parkin, the Fab comprising a Fab heavy chain comprising a heavy chain complementarity determining region 1 (HCDR1) comprising the amino acid sequence of SEQ ID NO: 4, a HCDR2 comprising the amino acid sequence of SEQ ID NO: 5, and a HCDR3 comprising the amino acid sequence of SEQ ID NO: 6, and a Fab light chain comprising a light chain complementarity determining region 1 (LCDR1) comprising the amino acid sequence of SEQ ID NO: 13, a LCDR2 comprising the amino acid sequence RAS, and a LCDR3 comprising the amino acid sequence of SEQ ID NO: 15.

[0218] The polynucleotide can encode a Fab that immunospecifically binds to Parkin, the Fab comprising a Fab heavy chain comprising a variable region comprising the amino acid sequence of SEQ ID NO: 7 and Fab light chain comprising a variable region comprising the amino acid sequence of SEQ ID NO: 16. The polynucleotide can encode a Fab heavy chain comprising a variable region comprising an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 7 and a Fab light chain comprising a variable region comprising an amino acid sequence with at least 90% sequence identity to the amino acid sequence of SEQ ID NO: 16. The polynucleotide can encode a Fab heavy chain comprising a variable region comprising an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 7 and a Fab light chain comprising a variable region comprising an amino acid sequence with at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 16.

[0219] The polynucleotide can encode a Fab that immunospecifically binds to Parkin, the Fab comprising a Fab heavy chain comprising a variable region comprising the amino acid sequence of SEQ ID NO: 9 and a Fab light chain comprising a variable region comprising the amino acid sequence of SEQ ID NO: 12. The polynucleotide can encode a Fab heavy chain comprising a variable region comprising an amino acid sequence with at least 90% sequence identity to the amino acid sequence of SEQ ID NO: 9 and Fab light chain comprising a variable region comprising an amino acid sequence with at least 90% sequence identity to the amino acid sequence of SEQ ID NO: 12. The polynucleotide can encode a Fab heavy chain comprising a variable region comprising an amino acid sequence with at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 9 and Fab light chain comprising a variable region comprising an amino acid sequence with at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 12. In some embodiments, in theAttorney Docket No. TP385350WO1 polynucleotide encoding a Fab that immunospecifically binds to Parkin, the polynucleotide comprises SEQ ID NO: 8 and SEQ ID NO: 11.

[0220] In some embodiments, the polynucleotide can encode an antibody comprising a Fab that immunospecifically binds to Parkin, the Fab comprising the amino acid sequence of SEQ ID NO: 3. The polynucleotide can encode a Fab comprising an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 3. The polynucleotide can encode a Fab comprising an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 3. In some embodiments, in the polynucleotide encoding a Fab that immunospecifically binds to Parkin, the polynucleotide comprises SEQ ID NO: 2.

[0221] In some embodiments, the polynucleotide can encode a Fab that immunospecifically binds to OCT4. The polynucleotide can encode a Fab that immunospecifically binds to OCT4, the Fab comprising a heavy chain complementarity determining region 1 (HCDR1) comprising the amino acid sequence of SEQ ID NO: 29, a HCDR2 comprising the amino acid sequence of SEQ ID NO: 30, and a HCDR3 comprising the amino acid sequence of SEQ ID NO: 31.

[0222] The polynucleotide can encode a Fab that immunospecifically binds to OCT4, the Fab comprising a Fab heavy chain comprising a variable region comprising the amino acid sequence of SEQ ID NO: 32. The polynucleotide can encode a Fab heavy chain comprising a variable region comprising an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 32. The polynucleotide can encode a Fab heavy chain comprising a variable region comprising an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 32.

[0223] The polynucleotide can encode a Fab that immunospecifically binds to OCT4, the Fab comprising a Fab heavy chain comprising a variable region comprising the amino acid sequence of SEQ ID NO: 34. The polynucleotide can encode a Fab heavy chain comprising a variable region comprising an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 34. The polynucleotide can encode a Fab heavy chain comprising a variable region comprising an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 34. In some embodiments, in the polynucleotide encoding a Fab that immunospecifically binds to OCT4, the polynucleotide comprises SEQ ID NO: 33.

[0224] The polynucleotide can encode a Fab that immunospecifically binds to OCT4, the Fab comprising a Fab light chain comprising a light chain complementarityAttorney Docket No. TP385350WO1 determining region 1 (LCDR1) comprising the amino acid sequence of SEQ ID NO: 37, a LCDR2 comprising the amino acid sequence GAS, and a LCDR3 comprising the amino acid sequence of SEQ ID NO: 39.

[0225] The polynucleotide can encode a Fab that immunospecifically binds to OCT4, the Fab comprising a Fab light chain comprising a variable region comprising the amino acid sequence of SEQ ID NO: 40. The polynucleotide can encode a Fab light chain comprising a variable region comprising an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 40. The polynucleotide can encode a Fab light chain comprising a variable region comprising an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 40.

[0226] The polynucleotide can encode a Fab that immunospecifically binds to OCT4, the Fab comprising a Fab light chain comprising the amino acid sequence of SEQ ID NO: 36. The polynucleotide can encode a Fab light chain comprising a variable region comprising an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 36. The polynucleotide can encode a Fab light chain comprising a variable region comprising an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 36. In some embodiments, in the polynucleotide encoding a Fab that immunospecifically binds to OCT4, the polynucleotide comprises SEQ ID NO: 35.

[0227] The polynucleotide can encode a Fab that immunospecifically binds to OCT4, the Fab comprising a Fab heavy chain comprising a heavy chain complementarity determining region 1 (HCDR1) comprising the amino acid sequence of SEQ ID NO: 29, a HCDR2 comprising the amino acid sequence of SEQ ID NO: 30, and a HCDR3 comprising the amino acid sequence of SEQ ID NO: 31, and a Fab light chain comprising a light chain complementarity determining region 1 (LCDR1) comprising the amino acid sequence of SEQ ID NO: 37, a LCDR2 comprising the amino acid sequence GAS, and a LCDR3 comprising the amino acid sequence of SEQ ID NO: 39.

[0228] The polynucleotide can encode a Fab that immunospecifically binds to OCT4, the Fab comprising a Fab heavy chain comprising a variable region comprising the amino acid sequence of SEQ ID NO: 32 and Fab light chain comprising a variable region comprising the amino acid sequence of SEQ ID NO: 40. The polynucleotide can encode a Fab heavy chain comprising a variable region comprising an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 32 and a Fab light chain comprising a variableAttorney Docket No. TP385350WO1 region comprising an amino acid sequence with at least 90% sequence identity to the amino acid sequence of SEQ ID NO: 40. The polynucleotide can encode a Fab heavy chain comprising a variable region comprising an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 32 and a Fab light chain comprising a variable region comprising an amino acid sequence with at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 40.

[0229] The polynucleotide can encode a Fab that immunospecifically binds to OCT4, the Fab comprising a Fab heavy chain comprising a variable region comprising the amino acid sequence of SEQ ID NO: 34 and a Fab light chain comprising a variable region comprising the amino acid sequence of SEQ ID NO: 36. The polynucleotide can encode a Fab heavy chain comprising a variable region comprising an amino acid sequence with at least 90% sequence identity to the amino acid sequence of SEQ ID NO: 34 and Fab light chain comprising a variable region comprising an amino acid sequence with at least 90% sequence identity to the amino acid sequence of SEQ ID NO: 36. The polynucleotide can encode a Fab heavy chain comprising a variable region comprising an amino acid sequence with at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 34 and Fab light chain comprising a variable region comprising an amino acid sequence with at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 36. In some embodiments, in the polynucleotide encoding a Fab that immunospecifically binds to OCT4, the polynucleotide comprises SEQ ID NO: 33 and SEQ ID NO: 35.

[0230] In some embodiments, the polynucleotide can encode an antibody comprising a Fab that immunospecifically binds to OCT4, the Fab comprising the amino acid sequence of SEQ ID NO: 28. The polynucleotide can encode a Fab comprising an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 28. The polynucleotide can encode a Fab comprising an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 28. In some embodiments, in the polynucleotide encoding a Fab that immunospecifically binds to OCT4, the polynucleotide comprises SEQ ID NO: 27.

[0231] In some embodiments, the polynucleotide can encode a Fab that immunospecifically binds to IDO1. The polynucleotide can encode a Fab that immunospecifically binds to IDO1, the Fab comprising a Fab heavy chain comprising a heavy chain complementarity determining region 1 (HCDR1) comprising the amino acidAttorney Docket No. TP385350WO1 sequence of SEQ ID NO: 66, a HCDR2 comprising the amino acid sequence of SEQ ID NO: 67, and a HCDR3 comprising the amino acid sequence of SEQ ID NO: 68.

[0232] The polynucleotide can encode a Fab that immunospecifically binds to IDO1, the Fab comprising a Fab heavy chain comprising a variable region comprising the amino acid sequence of SEQ ID NO: 64. The polynucleotide can encode a Fab heavy chain comprising a variable region comprising an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 64. The polynucleotide can encode a Fab heavy chain comprising a variable region comprising an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 64.

[0233] The polynucleotide can encode a Fab that immunospecifically binds IDO1, the Fab comprising a Fab light chain comprising a light chain complementarity determining region 1 (LCDR1) comprising the amino acid sequence of SEQ ID NO: 72, a LCDR2 comprising the amino acid sequence of YAS, and a LCDR3 comprising the amino acid sequence of SEQ ID NO: 73.

[0234] The polynucleotide can encode a Fab that immunospecifically binds IDO1, the Fab comprising a Fab light chain comprising a variable region comprising the amino acid sequence of SEQ ID NO: 71. The polynucleotide can encode a Fab light chain comprising a variable region comprising an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 71. The polynucleotide can encode a Fab light chain comprising a variable region comprising an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 71.

[0235] The polynucleotide can encode a Fab that immunospecifically binds to IDO1, the Fab comprising a Fab heavy chain comprising a heavy chain complementarity determining region 1 (HCDR1) comprising the amino acid sequence of SEQ ID NO: 66, a HCDR2 comprising the amino acid sequence of SEQ ID NO: 67, and a HCDR3 comprising the amino acid sequence of SEQ ID NO: 68, and a Fab light chain comprising a light chain complementarity determining region 1 (LCDR1) comprising the amino acid sequence of SEQ ID NO: 72, a LCDR2 comprising the amino acid sequence of YAS, and a LCDR3 comprising the amino acid sequence of SEQ ID NO: 73.

[0236] The polynucleotide can encode a Fab that immunospecifically binds to IDO1, the Fab comprising a Fab heavy chain comprising a variable region comprising the amino acid sequence of SEQ ID NO: 65 and a Fab light chain comprising a variable regionAttorney Docket No. TP385350WO1 comprising the amino acid sequence of SEQ ID NO: 71. The polynucleotide can encode a Fab heavy chain comprising a variable region comprising an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 65 and a Fab light chain comprising a variable region comprising an amino acid sequence with at least 90% sequence identity to the amino acid sequence of SEQ ID NO: 71. The polynucleotide can encode a Fab heavy chain comprising a variable region comprising an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 65 and a Fab light chain comprising a variable region comprising an amino acid sequence with at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 71. In some embodiments, in the polynucleotide encoding a Fab that immunospecifically binds to IDO1, the polynucleotide comprises SEQ ID NO: 64 and SEQ ID NO: 70.

[0237] In some embodiments, the polynucleotide can encode an antibody comprising a Fab that immunospecifically binds to IDO1, the Fab comprising the amino acid sequence of SEQ ID NO: 79. The polynucleotide can encode a Fab comprising an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 79. The polynucleotide can encode a Fab comprising an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 79. In some embodiments, in the polynucleotide encoding a Fab that immunospecifically binds to IDO1, the polynucleotide comprises SEQ ID NO: 78.

[0238] The polynucleotide can encode a Fab that immunospecifically binds to FLAG-tag, the Fab comprising a Fab heavy chain comprising a variable region comprising the amino acid sequence of SEQ ID NO: 162. The polynucleotide can encode a Fab heavy chain comprising a variable region comprising an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 162. The polynucleotide can encode a Fab heavy chain comprising a variable region comprising an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 162.

[0239] The polynucleotide can encode a Fab that immunospecifically binds FLAG-tag, the Fab comprising a Fab light chain comprising a light chain complementarity determining region 1 (LCDR1) comprising the amino acid sequence of SEQ ID NO: 168, a LCDR2 comprising the amino acid sequence of YAS, and a LCDR3 comprising the amino acid sequence of SEQ ID NO: 169.

[0240] The polynucleotide can encode a Fab that immunospecifically binds FLAG-tag, the Fab comprising a Fab light chain comprising a variable region comprising theAttorney Docket No. TP385350WO1 amino acid sequence of SEQ ID NO: 167. The polynucleotide can encode a Fab light chain comprising a variable region comprising an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 167. The polynucleotide can encode a Fab light chain comprising a variable region comprising an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 167.

[0241] The polynucleotide can encode a Fab that immunospecifically binds to FLAG-tag, the Fab comprising a Fab heavy chain comprising a heavy chain complementarity determining region 1 (HCDR1) comprising the amino acid sequence of SEQ ID NO: 163, a HCDR2 comprising the amino acid sequence of SEQ ID NO: 164, and a HCDR3 comprising the amino acid sequence of SEQ ID NO: 165, and a Fab light chain comprising a light chain complementarity determining region 1 (LCDR1) comprising the amino acid sequence of SEQ ID NO: 168, a LCDR2 comprising the amino acid sequence of YAS, and a LCDR3 comprising the amino acid sequence of SEQ ID NO: 169.

[0242] The polynucleotide can encode a Fab that immunospecifically binds to FLAG-tag, the Fab comprising a Fab heavy chain comprising a variable region comprising the amino acid sequence of SEQ ID NO: 162 and a Fab light chain comprising a variable region comprising the amino acid sequence of SEQ ID NO: 167. The polynucleotide can encode a Fab heavy chain comprising a variable region comprising an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 162 and a Fab light chain comprising a variable region comprising an amino acid sequence with at least 90% sequence identity to the amino acid sequence of SEQ ID NO: 167. The polynucleotide can encode a Fab heavy chain comprising a variable region comprising an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 162 and a Fab light chain comprising a variable region comprising an amino acid sequence with at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 167. In some embodiments, in the polynucleotide encoding a Fab that immunospecifically binds to FLAG-tag, the polynucleotide comprises SEQ ID NO: 181 and SEQ ID NO: 166.

[0243] In some embodiments, the polynucleotide can encode an antibody comprising a Fab that immunospecifically binds to FLAG-tag, the Fab comprising the amino acid sequence of SEQ ID NO: 175. The polynucleotide can encode a Fab comprising an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 175. The polynucleotide can encode a Fab comprising an amino acid sequence with at least 95%Attorney Docket No. TP385350WO1 sequence identity to SEQ ID NO: 175. In some embodiments, in the polynucleotide encoding a Fab that immunospecifically binds to FLAG-tag, the polynucleotide comprises SEQ ID NO: 174.

[0244] In some embodiments, the polynucleotide can encode a Fab that is a rabbit anti-mouse IgG secondary antibody (RAM). The polynucleotide can encode a Fab that is a RAM, the Fab comprising a Fab heavy chain comprising a heavy chain complementarity determining region 1 (HCDR1) comprising the amino acid sequence of SEQ ID NO: 127, a HCDR2 comprising the amino acid sequence of SEQ ID NO: 128, and a HCDR3 comprising the amino acid sequence of SEQ ID NO: 129.

[0245] The polynucleotide can encode a Fab that is a RAM, the Fab comprising a Fab heavy chain comprising a variable region comprising the amino acid sequence of SEQ ID NO: 126. The polynucleotide can encode a Fab heavy chain comprising a variable region comprising an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 126. The polynucleotide can encode a Fab heavy chain comprising a variable region comprising an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 126.

[0246] The polynucleotide can encode a Fab that is a RAM, the Fab comprising a Fab light chain comprising a light chain complementarity determining region 1 (LCDR1) comprising the amino acid sequence of SEQ ID NO: 132, a LCDR2 comprising the amino acid sequence of GAS, and a LCDR3 comprising the amino acid sequence of SEQ ID NO: 133.

[0247] The polynucleotide can encode a Fab that is a RAM, the Fab comprising a Fab light chain comprising a variable region comprising the amino acid sequence of SEQ ID NO: 131. The polynucleotide can encode a Fab light chain comprising a variable region comprising an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 131. The polynucleotide can encode a Fab light chain comprising a variable region comprising an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 131.

[0248] The polynucleotide can encode a Fab that is a RAM, the Fab comprising a Fab heavy chain comprising a heavy chain complementarity determining region 1 (HCDR1) comprising the amino acid sequence of SEQ ID NO: 127, a HCDR2 comprising the amino acid sequence of SEQ ID NO: 128, and a HCDR3 comprising the amino acid sequence of SEQ ID NO: 129, and a Fab light chain comprising a light chain complementarity determining region 1 (LCDR1) comprising the amino acid sequence of SEQ ID NO: 132, aAttorney Docket No. TP385350WO1 LCDR2 comprising the amino acid sequence of GAS, and a LCDR3 comprising the amino acid sequence of SEQ ID NO: 133.

[0249] The polynucleotide can encode a Fab that is a RAM, the Fab comprising a Fab heavy chain comprising a variable region comprising the amino acid sequence of SEQ ID NO: 126 and a Fab light chain comprising a variable region comprising the amino acid sequence of SEQ ID NO: 131. The polynucleotide can encode a Fab heavy chain comprising a variable region comprising an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 126 and a Fab light chain comprising a variable region comprising an amino acid sequence with at least 90% sequence identity to the amino acid sequence of SEQ ID NO: 131. The polynucleotide can encode a Fab heavy chain comprising a variable region comprising an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 126 and a Fab light chain comprising a variable region comprising an amino acid sequence with at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 131. In some embodiments, in the polynucleotide encoding a Fab that immunospecifically binds is a RAM, the polynucleotide comprises SEQ ID NO: 125 and SEQ ID NO: 130.

[0250] In some embodiments, the polynucleotide can encode an antibody comprising a Fab that is a RAM, the Fab comprising the amino acid sequence of SEQ ID NO: 139. The polynucleotide can encode a Fab comprising an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 139. The polynucleotide can encode a Fab comprising an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 139. In some embodiments, in the polynucleotide encoding a Fab that is a RAM, the polynucleotide comprises SEQ ID NO: 138.

[0251] In some embodiments, the polynucleotide can encode a Fab that immunospecifically binds to pERK1 / 2. The polynucleotide can encode a Fab that immunospecifically binds to pERK1 / 2, the Fab comprising a Fab heavy chain comprising a heavy chain complementarity determining region 1 (HCDR1) comprising the amino acid sequence of SEQ ID NO: 85, a HCDR2 comprising the amino acid sequence of SEQ ID NO: 86, and a HCDR3 comprising the amino acid sequence of SEQ ID NO: 87.

[0252] The polynucleotide can encode a Fab that immunospecifically binds to pERK1 / 2, the Fab comprising a Fab heavy chain comprising a variable region comprising the amino acid sequence of SEQ ID NO: 84. The polynucleotide can encode a Fab heavy chain comprising a variable region comprising an amino acid sequence with at least 90% sequenceAttorney Docket No. TP385350WO1 identity to SEQ ID NO: 84. The polynucleotide can encode a Fab heavy chain comprising a variable region comprising an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 84.

[0253] The polynucleotide can encode a Fab that immunospecifically binds pERK1 / 2, the Fab comprising a Fab light chain comprising a light chain complementarity determining region 1 (LCDR1) comprising the amino acid sequence of SEQ ID NO: 91, a LCDR2 comprising the amino acid sequence of KVS, and a LCDR3 comprising the amino acid sequence of SEQ ID NO: 92.

[0254] The polynucleotide can encode a Fab that immunospecifically binds pERK1 / 2, the Fab comprising a Fab light chain comprising a variable region comprising the amino acid sequence of SEQ ID NO: 90. The polynucleotide can encode a Fab light chain comprising a variable region comprising an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 90. The polynucleotide can encode a Fab light chain comprising a variable region comprising an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 90.

[0255] The polynucleotide can encode a Fab that immunospecifically binds to pERK1 / 2, the Fab comprising a Fab heavy chain comprising a heavy chain complementarity determining region 1 (HCDR1) comprising the amino acid sequence of SEQ ID NO: 85, a HCDR2 comprising the amino acid sequence of SEQ ID NO: 86, and a HCDR3 comprising the amino acid sequence of SEQ ID NO: 87, and a Fab light chain comprising a light chain complementarity determining region 1 (LCDR1) comprising the amino acid sequence of SEQ ID NO: 91, a LCDR2 comprising the amino acid sequence of KVS, and a LCDR3 comprising the amino acid sequence of SEQ ID NO: 92.

[0256] The polynucleotide can encode a Fab that immunospecifically binds to pERK1 / 2, the Fab comprising a Fab heavy chain comprising a variable region comprising the amino acid sequence of SEQ ID NO: 84 and a Fab light chain comprising a variable region comprising the amino acid sequence of SEQ ID NO: 90. The polynucleotide can encode a Fab heavy chain comprising a variable region comprising an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 84 and a Fab light chain comprising a variable region comprising an amino acid sequence with at least 90% sequence identity to the amino acid sequence of SEQ ID NO: 90. The polynucleotide can encode a Fab heavy chain comprising a variable region comprising an amino acid sequence with at least 95% sequenceAttorney Docket No. TP385350WO1 identity to SEQ ID NO: 84 and a Fab light chain comprising a variable region comprising an amino acid sequence with at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 90. In some embodiments, in the polynucleotide encoding a Fab that immunospecifically binds to pERK1 / 2, the polynucleotide comprises SEQ ID NO: 83 and SEQ ID NO: 89.

[0257] In some embodiments, the polynucleotide can encode an antibody comprising a Fab that immunospecifically binds to pERK1 / 2, the Fab comprising the amino acid sequence of SEQ ID NO: 98. The polynucleotide can encode a Fab comprising an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 98. The polynucleotide can encode a Fab comprising an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 98. In some embodiments, in the polynucleotide encoding a Fab that immunospecifically binds to pERK1 / 2, the polynucleotide comprises SEQ ID NO: 97.

[0258] In some embodiments, the polynucleotide can encode a Fab that immunospecifically binds to PSD-95. The polynucleotide can encode a Fab that immunospecifically binds to PSD-95, the Fab comprising a Fab heavy chain comprising a heavy chain complementarity determining region 1 (HCDR1) comprising the amino acid sequence of SEQ ID NO: 111, a HCDR2 comprising the amino acid sequence of SEQ ID NO: 112, and a HCDR3 comprising the amino acid sequence of SEQ ID NO: 113.

[0259] The polynucleotide can encode a Fab that immunospecifically binds to PSD-95, the Fab comprising a Fab heavy chain comprising a variable region comprising the amino acid sequence of SEQ ID NO: 110. The polynucleotide can encode a Fab heavy chain comprising a variable region comprising an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 110. The polynucleotide can encode a Fab heavy chain comprising a variable region comprising an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 110.

[0260] The polynucleotide can encode a Fab that immunospecifically binds PSD- 95, the Fab comprising a Fab light chain comprising a light chain complementarity determining region 1 (LCDR1) comprising the amino acid sequence of SEQ ID NO: 116, a LCDR2 comprising the amino acid sequence of DTS, and a LCDR3 comprising the amino acid sequence of SEQ ID NO: 117.

[0261] The polynucleotide can encode a Fab that immunospecifically binds PSD- 95, the Fab comprising a Fab light chain comprising a variable region comprising the aminoAttorney Docket No. TP385350WO1 acid sequence of SEQ ID NO: 115. The polynucleotide can encode a Fab light chain comprising a variable region comprising an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 115. The polynucleotide can encode a Fab light chain comprising a variable region comprising an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 115.

[0262] The polynucleotide can encode a Fab that immunospecifically binds to PSD-95, the Fab comprising a Fab heavy chain comprising a heavy chain complementarity determining region 1 (HCDR1) comprising the amino acid sequence of SEQ ID NO: 111, a HCDR2 comprising the amino acid sequence of SEQ ID NO: 112, and a HCDR3 comprising the amino acid sequence of SEQ ID NO: 113, and a Fab light chain comprising a light chain complementarity determining region 1 (LCDR1) comprising the amino acid sequence of SEQ ID NO: 116, a LCDR2 comprising the amino acid sequence of DTS, and a LCDR3 comprising the amino acid sequence of SEQ ID NO: 117.

[0263] The polynucleotide can encode a Fab that immunospecifically binds to PSD-95, the Fab comprising a Fab heavy chain comprising a variable region comprising the amino acid sequence of SEQ ID NO: 110 and a Fab light chain comprising a variable region comprising the amino acid sequence of SEQ ID NO: 115. The polynucleotide can encode a Fab heavy chain comprising a variable region comprising an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 110 and a Fab light chain comprising a variable region comprising an amino acid sequence with at least 90% sequence identity to the amino acid sequence of SEQ ID NO: 115. The polynucleotide can encode a Fab heavy chain comprising a variable region comprising an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 110 and a Fab light chain comprising a variable region comprising an amino acid sequence with at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 115. In some embodiments, in the polynucleotide encoding a Fab that immunospecifically binds to PSD-95, the polynucleotide comprises SEQ ID NO: 125 and SEQ ID NO: 130.

[0264] In some embodiments, the polynucleotide can encode an antibody comprising a Fab that immunospecifically binds to PSD-95, the Fab comprising the amino acid sequence of SEQ ID NO: 123. The polynucleotide can encode a Fab comprising an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 123. The polynucleotide can encode a Fab comprising an amino acid sequence with at least 95%Attorney Docket No. TP385350WO1 sequence identity to SEQ ID NO: 123. In some embodiments, in the polynucleotide encoding a Fab that immunospecifically binds to PSD-95, the polynucleotide comprises SEQ ID NO: 122.

[0265] In some embodiments, the polynucleotide can encode a Fab that immunospecifically binds to TIGIT. The polynucleotide can encode a Fab that immunospecifically binds to TIGIT, the Fab comprising a heavy chain complementarity determining region 1 (HCDR1) comprising the amino acid sequence of SEQ ID NO: 146, a HCDR2 comprising the amino acid sequence of SEQ ID NO: 147, and a HCDR3 comprising the amino acid sequence of SEQ ID NO: 148

[0266] The polynucleotide can encode a Fab that immunospecifically binds to TIGIT, the Fab comprising a Fab heavy chain comprising a variable region comprising the amino acid sequence of SEQ ID NO: 145. The polynucleotide can encode a Fab heavy chain comprising a variable region comprising an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 145. The polynucleotide can encode a Fab heavy chain comprising a variable region comprising an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 145.

[0267] The polynucleotide can encode a Fab that immunospecifically binds to TIGIT, the Fab comprising a Fab light chain comprising a light chain complementarity determining region 1 (LCDR1) comprising the amino acid sequence of SEQ ID NO: 151, a LCDR2 comprising the amino acid sequence of SAS, and a LCDR3 comprising the amino acid sequence of SEQ ID NO: 152.

[0268] The polynucleotide can encode a Fab that immunospecifically binds to TIGIT, the Fab comprising a Fab light chain comprising a variable region comprising the amino acid sequence of SEQ ID NO: 150. The polynucleotide can encode a Fab light chain comprising a variable region comprising an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 150. The polynucleotide can encode a Fab light chain comprising a variable region comprising an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 150.

[0269] The polynucleotide can encode a Fab that immunospecifically binds to TIGIT, the Fab comprising a Fab heavy chain comprising a heavy chain complementarity determining region 1 (HCDR1) comprising the amino acid sequence of SEQ ID NO: 146, a HCDR2 comprising the amino acid sequence of SEQ ID NO: 147, and a HCDR3 comprisingAttorney Docket No. TP385350WO1 the amino acid sequence of SEQ ID NO: 148, and a Fab light chain comprising a light chain complementarity determining region 1 (LCDR1) comprising the amino acid sequence of SEQ ID NO: 151, a LCDR2 comprising the amino acid sequence of SAS, and a LCDR3 comprising the amino acid sequence of SEQ ID NO: 152.

[0270] The polynucleotide can encode a Fab that immunospecifically binds to TIGIT, the Fab comprising a Fab heavy chain comprising a variable region comprising the amino acid sequence of SEQ ID NO: 145 and a Fab light chain comprising a variable region comprising the amino acid sequence of SEQ ID NO: 150. The polynucleotide can encode a Fab heavy chain comprising a variable region comprising an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 145 and a Fab light chain comprising a variable region comprising an amino acid sequence with at least 90% sequence identity to the amino acid sequence of SEQ ID NO: 150. The polynucleotide can encode a Fab heavy chain comprising a variable region comprising an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 145 and a Fab light chain comprising a variable region comprising an amino acid sequence with at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 150. In some embodiments, in the polynucleotide encoding a Fab that immunospecifically binds to TIGIT, the polynucleotide comprises SEQ ID NO: 144 and SEQ ID NO: 149.

[0271] In some embodiments, the polynucleotide can encode a Fab that immunospecifically binds to TIGIT, the Fab comprising the amino acid sequence of SEQ ID NO: 158. The polynucleotide can encode a Fab comprising an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 158. The polynucleotide can encode a Fab comprising an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 158. In some embodiments, in the polynucleotide encoding a Fab that immunospecifically binds to TIGIT, the polynucleotide comprises SEQ ID NO: 157.

[0272] The polynucleotide can encode any antibody disclosed herein wherein the antibody has the isotype IgG.

[0273] Disclosed herein are vectors comprising any of the herein described nucleic acid molecules that comprise polynucleotides encoding isolated polypeptides or antibodies. Disclosed herein are also host cells comprising any of the herein disclosed vectors. Suitable host cells include eukaryotic cells and prokaryotic cells.Attorney Docket No. TP385350WO1

[0274] Disclosed herein are methods of producing an isolated polypeptide or antibody, the methods comprising culturing a host cell comprising any of the herein disclosed vectors under conditions suitable for expression of the isolated polypeptide or antibody and purifying the isolated polypeptide or antibody.

[0275] Disclosed herein are antibodies comprising the isolated polypeptide comprising the amino acid sequence of Formula I: Z-(X-Yn-Zr)p-X, wherein Z is a hinge region, X is a CH2 domain, Y is a linker, n is an integer of 1 or greater, p is an integer of 2 or greater, and r is an integer of 0 or 1 linked to a Fab domain.

[0276] The hinge (Z) region can comprise the amino acid sequence of SEQ ID NO: 18. The hinge (Z) can comprise an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 18. The hinge (Z) can comprise an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 18. The hinge (Z) can comprise the amino acid sequence of SEQ ID NO: 48. The hinge (Z) can comprise an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 48. The hinge (Z) can comprise an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 48. The hinge (Z) can comprise the amino acid sequence of SEQ ID NO: 102. The hinge (Z) can comprise an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 102. The hinge (Z) can comprise an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 102.

[0277] The CH2 domain (X) can comprise the amino acid sequence of SEQ ID NO: 19. The CH2 domain (X) can comprise an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 19. The CH2 domain (X) can comprise an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 19. The CH2 domain (X) can comprise the amino acid sequence of SEQ ID NO: 49. The CH2 domain (X) can comprise an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 49. The CH2 domain (X) can comprise an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 49. The CH2 domain (X) can comprise the amino acid sequence of SEQ ID NO: 104. The CH2 domain (X) can comprise an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 104. The CH2 domain (X) can comprise an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 104.

[0278] In some embodiments, the antibody comprising the isolated polypeptide can comprise one or more linkers (Y) comprising the amino acid sequence of GS, GGS, SEQ ID NO: 14, SEQ ID NO: 38, SEQ ID NO: 43, or SEQ ID NO: 44. The one or more linkersAttorney Docket No. TP385350WO1 (Y) can comprise GS. The one or more linkers (Y) can comprise GGS. The one or more linkers (Y) can comprise SEQ ID NO: 14. The one or more linkers (Y) can comprise SEQ ID NO: 38. The one or more linkers can comprise SEQ ID NO: 43. The one or more linkers (Y) can comprise a sequence with at least 90% sequence identity to SEQ ID NO: 43. The one or more linkers (Y) can comprise SEQ ID NO: 44. The one or more linkers (Y) can comprise a sequence with at least 90% sequence identity to SEQ ID NO: 44.

[0279] The isolated polypeptide can comprise one linker (Y), such that n is 1. The isolated polypeptide can comprise two linkers (Y), such that n is 2. The isolated polypeptide can comprise three linkers (Y), such that n is 3. The isolated polypeptide can comprise four linkers (Y), such that n is 4. The isolated polypeptide can comprise five linkers (Y), such that n is 5.

[0280] In some embodiments, the antibody comprising the isolated polypeptide can comprise two or more regions comprising a CH2-linker region ((X-Yn)p), such that p is 2 or greater. The isolated polypeptide can comprise two CH2-linker regions ((X-Yn)p), such that p is 2. The isolated polypeptide can comprise three CH2-linker regions ((X-Yn)p), such that p is 3. The isolated polypeptide can comprise four CH2-linker regions ((X-Yn)p), such that p is 4. The isolated polypeptide can comprise five CH2-linker regions ((X-Yn)p), such that p is 5.

[0281] In some embodiments, the antibody comprising the isolated polypeptide can comprise two or more regions comprising a CH2-linker-hinge region ((X-Yn-Zr)p), such that p is 2 or greater. The isolated polypeptide can comprise two CH2-linker-hinge regions ((X-Yn-Zr)p), such that p is 2. The isolated polypeptide can comprise three CH2-linker-hinge regions ((X-Yn-Zr)p), such that p is 3. The isolated polypeptide can comprise four CH2- linker-hinge regions ((X-Yn-Zr)p), such that p is 4. The isolated polypeptide can comprise five CH2-linker-hinge regions ((X-Yn-Zr)p), such that p is 5.

[0282] In some embodiments, the antibody comprising the isolated polypeptide comprises, consists essentially of, or consists of (X-Yn-Zr)p, wherein Z (hinge) comprises the amino acid sequence of SEQ ID NO: 18, X (CH2 domain) comprises the amino acid sequence of SEQ ID NO: 19, Y (linker) comprises the amino acid sequence of SEQ ID NO: 44, wherein n is 1, r is 0 or 1, and p is 2.

[0283] In some embodiments, the antibody comprising the isolated polypeptide comprises, consists essentially of, or consists of (X-Yn-Zr)p, wherein Z (hinge) comprises theAttorney Docket No. TP385350WO1 amino acid sequence of SEQ ID NO: 48, X (CH2 domain) comprises the amino acid sequence of SEQ ID NO: 49, Y (linker) comprises the amino acid sequence of SEQ ID NO: 44, wherein n is 1, r is 0 or 1, and p is 2.

[0284] In some embodiments, the antibody comprising the isolated polypeptide comprises, consists essentially of, or consists of (X-Yn-Zr)p, wherein Z (hinge) comprises the amino acid sequence of SEQ ID NO: 102, X (CH2 domain) comprises the amino acid sequence of SEQ ID NO: 104, Y (linker) comprises the amino acid sequence of SEQ ID NO: 44, wherein n is 1, r is 0 or 1, and p is 2.

[0285] In some embodiments, the antibody comprising the isolated polypeptide comprises, consists essentially of, or consists of (X-Yn-Zr)p, wherein Z (hinge) comprises the amino acid sequence of SEQ ID NO: 102, X (CH2 domain) comprises the amino acid sequence of SEQ ID NO: 104, Y (linker) comprises the amino acid sequence of SEQ ID NO: 45, wherein n is 1, r is 0 or 1, and p is 2.

[0286] In some embodiments, the antibody comprising the isolated polypeptide comprises two additional CH2 domains N-terminal to the Fc domain connected via a hinge and optionally, one or more linkers. The isolated polypeptide comprising two additional CH2 domains N-terminal to the Fc domain connected via a hinge and optionally one or more linkers can comprise the amino acid sequence of SEQ ID NO: 52 or SEQ ID NO: 54. In some embodiments, the isolated polypeptide comprising a CH3 domain comprises SEQ ID NO: 52. The isolated polypeptide comprising two additional CH2 domains N-terminal to the Fc domain connected via a hinge and optionally one or more linkers can comprise an amino acid sequence with at least 90% identity to SEQ ID NO: 52. The isolated polypeptide comprising two additional CH2 domains N-terminal to the Fc domain connected via a hinge and optionally one or more linkers can comprise an amino acid sequence with at least 95% identity to SEQ ID NO: 52.

[0287] In some embodiments, the antibody comprising the isolated polypeptide comprises two additional CH2 domains N-terminal to the Fc domain connected via a hinge and optionally one or more linkers and comprises “LFLEPS” mutations within the lower hinge and CH2 domains. The “LFLEPS” mutations can comprise the following amino acid substitutions: L234F, L235E and P331S. The isolated polypeptide comprising two additional CH2 domains N-terminal to the Fc domain connected via a hinge and optionally one or more linkers and comprises “LFLEPS” mutations within the lower hinge and CH2Attorney Docket No. TP385350WO1 domains comprises SEQ ID NO: 54. The isolated polypeptide comprising two additional CH2 domains N-terminal to the Fc domain connected via a hinge and optionally one or more linkers and comprises “LFLEPS” mutations within the lower hinge and CH2 domains can comprise an amino acid sequence with at least 90% identity to SEQ ID NO: 54. The isolated polypeptide comprising two additional CH2 domains N-terminal to the Fc domain connected via a hinge and optionally one or more linkers and comprises “LFLEPS” mutations within the lower hinge and CH2 domains can comprise an amino acid sequence with at least 95% identity to SEQ ID NO: 54.

[0288] In some embodiments, the antibodies comprising the isolated peptide disclosed herein have a reduction of background signal of at least 1.5-fold, compared to a corresponding wild type antibody. In some embodiments, the reduction of background signal is at least 1.5-fold, at least 2-fold, at least 2.5-fold, at least 3-fold, at least 3.5-fold, at least 4- fold, at least 4.5-fold or at least 5-fold compared to a corresponding wild type antibody.

[0289] In some embodiments, the antibodies comprising any isolated peptide disclosed herein have an increase in sensitivity or signal enhancement of at least 1.5-fold compared to a corresponding wild type antibody. In some embodiments, the increase in sensitivity is at least 1.5-fold, at least 2-fold, at least 2.5-fold, at least 3-fold, at least 3.5-fold, at least 4-fold, at least 4.5-fold, at least 5-fold, at least 5.5-fold, at least 6-fold, at least 6.5- fold, at least 7-fold, at least 7.5-fold, at least 8-fold, at least 8.5-fold, at least 9-fold, at least 9.5-fold, or at least 10-fold compared to a corresponding wild type antibody.

[0290] Disclosed herein are antibodies that immunospecifically bind to Parkin, wherein the antibodies comprise, consist essentially of, or consist of a heavy chain (HC) and a light chain (LC) that pair to form a binding domain comprising a heavy chain complementarity determining region 1 (HCDR1) comprising the amino acid sequence of SEQ ID NO: 4, a HCDR2 comprising the amino acid sequence of SEQ ID NO: 5, a HCDR3 comprising the amino acid sequence of SEQ ID NO: 6, a light chain complementarity determining region 1 (LCDR1) comprising the amino acid sequence of SEQ ID NO: 13, a LCDR2 comprising the amino acid sequence RAS, and a LCDR3 comprising the amino acid sequence of SEQ ID NO: 15, and wherein the heavy chain further comprises a first CH2 domain, a second CH2 domain, and a third CH2 domain, wherein the first CH2 domain and the second CH2 domain are connected by a first linker and an optional first hinge, and wherein the second CH2 domain and the third CH2 domain are connected by a second linkerAttorney Docket No. TP385350WO1 and an optional second hinge. In some embodiments, the antibodies that immunospecifically bind to Parkin further comprise a CH3 domain linked to the C-terminus of the third CH2 domain.

[0291] The first CH2 domain, the second CH2 domain, and the third CH2 domain can each comprise the amino acid sequence of SEQ ID NO: 19 or SEQ ID NO: 50. In some embodiments, the first CH2 domain, the second CH2 domain, and the third CH2 domain each comprise the amino acid sequence of SEQ ID NO: 19. The first CH2 domain, the second CH2 domain, and the third CH2 domain can each comprise an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 19. The first CH2 domain, the second CH2 domain, and the third CH2 domain can each comprise an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 19.

[0292] In some embodiments, the first CH2 domain, the second CH2 domain, and the third CH2 domain can each comprise the amino acid sequence of SEQ ID NO: 49. The first CH2 domain, the second CH2 domain, and the third CH2 domain can each comprise an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 49. The first CH2 domain, the second CH2 domain, and the third CH2 domain can each comprise an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 49.

[0293] The first hinge and the second hinge can each comprise the amino acid sequence of SEQ ID NO: 18 or SEQ ID NO: 48. In some embodiments, the first hinge and the second hinge can each comprise the amino acid sequence of SEQ ID NO: 18. The first hinge and the second hinge can each comprise an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 18. The first hinge and the second hinge can each comprise an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 18.

[0294] In some embodiments, the first hinge and the second hinge can each comprise the amino acid sequence of SEQ ID NO: 48. The first hinge and the second hinge can each comprise an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 48. The first hinge and the second hinge can each comprise an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 48.

[0295] The first linker and the second linker can each comprise the amino acid sequence of GS, GGS, SEQ ID NO: 14, SEQ ID NO: 38, SEQ ID NO: 43, or SEQ ID NO: 44. The first and the second linkers can comprise GS. The first and the second linkers can comprise GGS. The first and the second linkers can comprise SEQ ID NO: 14. The first andAttorney Docket No. TP385350WO1 the second linkers can comprise SEQ ID NO: 38. The first and the second linkers can comprise SEQ ID NO: 43. The first and the second linkers can comprise a sequence with at least 90% sequence identity to SEQ ID NO: 43. The first and the second linkers can comprise SEQ ID NO: 44. The first and the second linkers can comprise a sequence with at least 90% sequence identity to SEQ ID NO: 44.

[0296] In some embodiments, the antibody comprises a CH3 domain comprising the amino acid sequence of SEQ ID NO: 20. The antibody can comprise a CH3 domain having an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 20. The antibody can comprise a CH3 domain having an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 20.

[0297] In some embodiments, the antibody comprises a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 7 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 16. The antibody can comprise a heavy chain variable region having an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 7 and a light chain variable region having an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 16. The antibody can comprise a heavy chain variable region having an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 7 and a light chain variable region having an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 16.

[0298] In some embodiments, the antibody comprises a heavy chain having a polypeptide sequence of SEQ ID NO: 3 and a light chain having the polypeptide sequence of SEQ ID NO: 12. The antibody can comprise a heavy chain having an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 3 and a light chain having an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 12. The antibody can comprise a heavy chain having an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 3 and a light chain having an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 12.

[0299] In some embodiments, the antibody comprises a heavy chain that is encoded by a polynucleotide sequence of SEQ ID NO: 2 and a light chain that is encoded by a polynucleotide sequence of SEQ ID NO: 11.

[0300] Disclosed herein are antibodies that immunospecifically bind to OCT4, wherein the antibodies comprise, consist essentially of, or consist a heavy chain (HC) and aAttorney Docket No. TP385350WO1 light chain (LC) that pair to form a binding domain comprising a heavy chain complementarity determining region 1 (HCDR1) comprising the amino acid sequence of SEQ ID NO: 29, a HCDR2 comprising the amino acid sequence of SEQ ID NO: 30, and a HCDR3 comprising the amino acid sequence of SEQ ID NO: 31, a light chain complementarity determining region 1 (LCDR1) comprising the amino acid sequence of SEQ ID NO: 37, a LCDR2 comprising the amino acid sequence GAS, and a LCDR3 comprising the amino acid sequence of SEQ ID NO: 39, and wherein the heavy chain further comprises a first CH2 domain, a second CH2 domain, and a third CH2 domain, wherein the first CH2 domain and the second CH2 domain are connected by a first linker and an optional first hinge, and wherein the second CH2 domain and the third CH2 domain are connected by a second linker and an optional second hinge. In some embodiments, the antibodies that immunospecifically bind to OCT4 further comprise a CH3 domain linked to the C-terminus of the third CH2 domain.

[0301] The first CH2 domain, the second CH2 domain, and the third CH2 domain can each comprise the amino acid sequence of SEQ ID NO: 19 or SEQ ID NO: 50. In some embodiments, the first CH2 domain, the second CH2 domain, and the third CH2 domain can each comprise the amino acid sequence of SEQ ID NO: 19. The first CH2 domain, the second CH2 domain, and the third CH2 domain can each comprise an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 19. The first CH2 domain, the second CH2 domain, and the third CH2 domain can each comprise an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 19.

[0302] In some embodiments, the first CH2 domain, the second CH2 domain, and the third CH2 domain can each comprise the amino acid sequence of SEQ ID NO: 49. The first CH2 domain, the second CH2 domain, and the third CH2 domain can each comprise an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 49. The first CH2 domain, the second CH2 domain, and the third CH2 domain can each comprise an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 49.

[0303] The first hinge and the second hinge can each comprise the amino acid sequence of SEQ ID NO: 18 or SEQ ID NO: 48. In some embodiments, the first hinge and the second hinge each comprise the amino acid sequence of SEQ ID NO: 18. The first hinge and the second hinge can each comprise an amino acid sequence with at least 90% sequenceAttorney Docket No. TP385350WO1 identity to SEQ ID NO: 18. The first hinge and the second hinge can each comprise an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 18.

[0304] In some embodiments, the first hinge and the second hinge can each comprise the amino acid sequence of SEQ ID NO: 48. The first hinge and the second hinge can each comprise an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 48. The first hinge and the second hinge can each comprise an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 48.

[0305] The first linker and the second linker can each comprise the amino acid sequence of GS, GGS, SEQ ID NO: 14, SEQ ID NO: 38, SEQ ID NO: 43, or SEQ ID NO: 44. The first and the second linkers can comprise GS. The first and the second linkers can comprise GGS. The first and the second linkers can comprise SEQ ID NO: 14. The first and the second linkers can comprise SEQ ID NO: 38. The first and the second linkers can comprise SEQ ID NO: 43. The first and the second linkers can comprise a sequence with at least 90% sequence identity to SEQ ID NO: 43. The first and the second linkers can comprise SEQ ID NO: 44. The first and the second linkers can comprise a sequence with at least 90% sequence identity to SEQ ID NO: 44.

[0306] In some embodiments, the antibody comprises a CH3 domain comprising the amino acid sequence of SEQ ID NO: 20. The antibody can comprise a CH3 domain having an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 20. The antibody can comprise a CH3 domain having an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 20.

[0307] In some embodiments, the antibody comprises a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 32 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 40. The antibody can comprise a heavy chain variable region having an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 32 and a light chain variable region having an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 40. The antibody can comprise a heavy chain variable region having an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 32 and a light chain variable region having an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 40.

[0308] In some embodiments, the antibody comprises a heavy chain having a polypeptide sequence of SEQ ID NO: 28 and a light chain having the polypeptide sequenceAttorney Docket No. TP385350WO1 of SEQ ID NO: 36. The antibody can comprise a heavy chain having an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 28 and a light chain having an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 36. The antibody can comprise a heavy chain having an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 28 and a light chain having an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 36.

[0309] In some embodiments, the antibody comprises a heavy chain that is encoded by a polynucleotide sequence of SEQ ID NO: 27 and a light chain that is encoded by a polynucleotide sequence of SEQ ID NO: 35.

[0310] Disclosed herein are antibodies that immunospecifically bind to IDO1, wherein the antibodies comprise, consist essentially of, or consist of a heavy chain (HC) and a light chain (LC) that pair to form a binding domain comprising a heavy chain complementarity determining region 1 (HCDR1) comprising the amino acid sequence of SEQ ID NO: 66, a HCDR2 comprising the amino acid sequence of SEQ ID NO: 67, a HCDR3 comprising the amino acid sequence of SEQ ID NO: 68, a light chain complementarity determining region 1 (LCDR1) comprising the amino acid sequence of SEQ ID NO: 72, a LCDR2 comprising the amino acid sequence of YAS, and a LCDR3 comprising the amino acid sequence of SEQ ID NO: 73, and wherein the heavy chain further comprises a first CH2 domain, a second CH2 domain, and a third CH2 domain, wherein the first CH2 domain and the second CH2 domain are connected by a first linker and an optional first hinge, and wherein the second CH2 domain and the third CH2 domain are connected by a second linker and an optional second hinge. In some embodiments, the antibodies that immunospecifically bind to IDO1 further comprise a CH3 domain linked to the C-terminus of the third CH2 domain.

[0311] The first CH2 domain, the second CH2 domain, and the third CH2 domain can each comprise the amino acid sequence of SEQ ID NO: 19 or SEQ ID NO: 49. In some embodiments, the first CH2 domain, the second CH2 domain, and the third CH2 domain each comprise the amino acid sequence of SEQ ID NO: 19. The first CH2 domain, the second CH2 domain, and the third CH2 domain can each comprise an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 19. The first CH2 domain, the second CH2 domain, and the third CH2 domain can each comprise an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 19.Attorney Docket No. TP385350WO1

[0312] In some embodiments, the first CH2 domain, the second CH2 domain, and the third CH2 domain can each comprise the amino acid sequence of SEQ ID NO: 49. The first CH2 domain, the second CH2 domain, and the third CH2 domain can each comprise an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 49. The first CH2 domain, the second CH2 domain, and the third CH2 domain can each comprise an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 49.

[0313] The first hinge and the second hinge can each comprise the amino acid sequence of SEQ ID NO: 18 or SEQ ID NO: 48. In some embodiments, the first hinge and the second hinge can each comprise the amino acid sequence of SEQ ID NO: 18. The first hinge and the second hinge can each comprise an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 18. The first hinge and the second hinge can each comprise an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 18.

[0314] In some embodiments, the first hinge and the second hinge can each comprise the amino acid sequence of SEQ ID NO: 48. The first hinge and the second hinge can each comprise an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 48. The first hinge and the second hinge can each comprise an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 48.

[0315] The first linker and the second linker can each comprise the amino acid sequence of GS, GGS, SEQ ID NO: 14, SEQ ID NO: 38, SEQ ID NO: 43, or SEQ ID NO: 44. The first and the second linkers can comprise GS. The first and the second linkers can comprise GGS. The first and the second linkers can comprise SEQ ID NO: 14. The first and the second linkers can comprise SEQ ID NO: 38. The first and the second linkers can comprise SEQ ID NO: 43. The first and the second linkers can comprise a sequence with at least 90% sequence identity to SEQ ID NO: 43. The first and the second linkers can comprise SEQ ID NO: 44. The first and the second linkers can comprise a sequence with at least 90% sequence identity to SEQ ID NO: 44.

[0316] In some embodiments, the antibody comprises a CH3 domain comprising the amino acid sequence of SEQ ID NO: 20. The antibody can comprise a CH3 domain having an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 20. The antibody can comprise a CH3 domain having an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 20.Attorney Docket No. TP385350WO1

[0317] In some embodiments, the antibody comprises a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 65 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 71. The antibody can comprise a heavy chain variable region having an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 65 and a light chain variable region having an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 71. The antibody can comprise a heavy chain variable region having an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 65 and a light chain variable region having an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 71.

[0318] In some embodiments, the antibody comprises a heavy chain having a polypeptide sequence of SEQ ID NO: 75 and a light chain having the polypeptide sequence of SEQ ID NO: 77. The antibody can comprise a heavy chain having an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 75 and a light chain having an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 77. The antibody can comprise a heavy chain having an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 75 and a light chain having an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 77.

[0319] In some embodiments, the antibody comprises a heavy chain that is encoded by a polynucleotide sequence of SEQ ID NO: 74 and a light chain that is encoded by a polynucleotide sequence of SEQ ID NO: 76.

[0320] Disclosed herein are antibodies that immunospecifically bind to FLAG-tag, wherein the antibodies comprise, consist essentially of, or consist of a heavy chain (HC) and a light chain (LC) that pair to form a binding domain comprising a heavy chain complementarity determining region 1 (HCDR1) comprising the amino acid sequence of SEQ ID NO: 163, a HCDR2 comprising the amino acid sequence of SEQ ID NO: 164, a HCDR3 comprising the amino acid sequence of SEQ ID NO: 165, a light chain complementarity determining region 1 (LCDR1) comprising the amino acid sequence of SEQ ID NO: 168, a LCDR2 comprising the amino acid sequence of YAS, and a LCDR3 comprising the amino acid sequence of SEQ ID NO: 169, and wherein the heavy chain further comprises a first CH2 domain, a second CH2 domain, and a third CH2 domain, wherein the first CH2 domain and the second CH2 domain are connected by a first linker and an optional first hinge, and wherein the second CH2 domain and the third CH2 domain areAttorney Docket No. TP385350WO1 connected by a second linker and an optional second hinge. In some embodiments, the antibodies that immunospecifically bind to FLAG-tag further comprise a CH3 domain linked to the C-terminus of the third CH2 domain.

[0321] The first CH2 domain, the second CH2 domain, and the third CH2 domain can each comprise the amino acid sequence of SEQ ID NO: 19 or SEQ ID NO: 49. In some embodiments, the first CH2 domain, the second CH2 domain, and the third CH2 domain each comprise the amino acid sequence of SEQ ID NO: 19. The first CH2 domain, the second CH2 domain, and the third CH2 domain can each comprise an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 19. The first CH2 domain, the second CH2 domain, and the third CH2 domain can each comprise an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 19.

[0322] In some embodiments, the first CH2 domain, the second CH2 domain, and the third CH2 domain can each comprise the amino acid sequence of SEQ ID NO: 49. The first CH2 domain, the second CH2 domain, and the third CH2 domain can each comprise an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 49. The first CH2 domain, the second CH2 domain, and the third CH2 domain can each comprise an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 49.

[0323] The first hinge and the second hinge can each comprise the amino acid sequence of SEQ ID NO: 18 or SEQ ID NO: 48. In some embodiments, the first hinge and the second hinge can each comprise the amino acid sequence of SEQ ID NO: 18. The first hinge and the second hinge can each comprise an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 18. The first hinge and the second hinge can each comprise an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 18.

[0324] In some embodiments, the first hinge and the second hinge can each comprise the amino acid sequence of SEQ ID NO: 48. The first hinge and the second hinge can each comprise an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 48. The first hinge and the second hinge can each comprise an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 48.

[0325] The first linker and the second linker can each comprise the amino acid sequence of GS, GGS, SEQ ID NO: 14, SEQ ID NO: 38, SEQ ID NO: 43, or SEQ ID NO: 44. The first and the second linkers can comprise GS. The first and the second linkers can comprise GGS. The first and the second linkers can comprise SEQ ID NO: 14. The first andAttorney Docket No. TP385350WO1 the second linkers can comprise SEQ ID NO: 38. The first and the second linkers can comprise SEQ ID NO: 43. The first and the second linkers can comprise a sequence with at least 90% sequence identity to SEQ ID NO: 43. The first and the second linkers can comprise SEQ ID NO: 44. The first and the second linkers can comprise a sequence with at least 90% sequence identity to SEQ ID NO: 44.

[0326] In some embodiments, the antibody comprises a CH3 domain comprising the amino acid sequence of SEQ ID NO: 20. The antibody can comprise a CH3 domain having an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 20. The antibody can comprise a CH3 domain having an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 20.

[0327] In some embodiments, the antibody comprises a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 162 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 167. The antibody can comprise a heavy chain variable region having an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 162 and a light chain variable region having an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 167. The antibody can comprise a heavy chain variable region having an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 162 and a light chain variable region having an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 167.

[0328] In some embodiments, the antibody comprises a heavy chain having a polypeptide sequence of SEQ ID NO: 171 and a light chain having the polypeptide sequence of SEQ ID NO: 173. The antibody can comprise a heavy chain having an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 171 and a light chain having an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 173. The antibody can comprise a heavy chain having an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 171 and a light chain having an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 173.

[0329] In some embodiments, the antibody comprises a heavy chain that is encoded by a polynucleotide sequence of SEQ ID NO: 170 and a light chain that is encoded by a polynucleotide sequence of SEQ ID NO: 172.

[0330] Disclosed herein are rabbit anti-mouse IgG secondary antibodies (RAM), wherein the antibodies comprise, consist essentially of, or consist of a heavy chain (HC) andAttorney Docket No. TP385350WO1 a light chain (LC) that pair to form a binding domain comprising a heavy chain complementarity determining region 1 (HCDR1) comprising the amino acid sequence of SEQ ID NO: 127, a HCDR2 comprising the amino acid sequence of SEQ ID NO: 128, a HCDR3 comprising the amino acid sequence of SEQ ID NO: 129, a light chain complementarity determining region 1 (LCDR1) comprising the amino acid sequence of SEQ ID NO: 132, a LCDR2 comprising the amino acid sequence of GAS, and a LCDR3 comprising the amino acid sequence of SEQ ID NO: 133, and wherein the heavy chain further comprises a first CH2 domain, a second CH2 domain, and a third CH2 domain, wherein the first CH2 domain and the second CH2 domain are connected by a first linker and an optional first hinge, and wherein the second CH2 domain and the third CH2 domain are connected by a second linker and an optional second hinge. In some embodiments, the rabbit anti-mouse IgG secondary antibodies further comprise a CH3 domain linked to the C- terminus of the third CH2 domain.

[0331] The first CH2 domain, the second CH2 domain, and the third CH2 domain can each comprise the amino acid sequence of SEQ ID NO: 19 or SEQ ID NO: 49. In some embodiments, the first CH2 domain, the second CH2 domain, and the third CH2 domain each comprise the amino acid sequence of SEQ ID NO: 19. The first CH2 domain, the second CH2 domain, and the third CH2 domain can each comprise an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 19. The first CH2 domain, the second CH2 domain, and the third CH2 domain can each comprise an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 19.

[0332] In some embodiments, the first CH2 domain, the second CH2 domain, and the third CH2 domain can each comprise the amino acid sequence of SEQ ID NO: 49. The first CH2 domain, the second CH2 domain, and the third CH2 domain can each comprise an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 49. The first CH2 domain, the second CH2 domain, and the third CH2 domain can each comprise an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 49.

[0333] The first hinge and the second hinge can each comprise the amino acid sequence of SEQ ID NO: 18 or SEQ ID NO: 48. In some embodiments, the first hinge and the second hinge can each comprise the amino acid sequence of SEQ ID NO: 18 The first hinge and the second hinge can each comprise an amino acid sequence with at least 90%Attorney Docket No. TP385350WO1 sequence identity to SEQ ID NO: 18. The first hinge and the second hinge can each comprise an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 18.

[0334] In some embodiments, the first hinge and the second hinge can each comprise the amino acid sequence of SEQ ID NO: 48. The first hinge and the second hinge can each comprise an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 48. The first hinge and the second hinge can each comprise an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 48.

[0335] The first linker and the second linker can each comprise the amino acid sequence of GS, GGS, SEQ ID NO: 14, SEQ ID NO: 38, SEQ ID NO: 43, or SEQ ID NO: 44. The first and the second linkers can comprise GS. The first and the second linkers can comprise GGS. The first and the second linkers can comprise SEQ ID NO: 14. The first and the second linkers can comprise SEQ ID NO: 38. The first and the second linkers can comprise SEQ ID NO: 43. The first and the second linkers can comprise a sequence with at least 90% sequence identity to SEQ ID NO: 43. The first and the second linkers can comprise SEQ ID NO: 44. The first and the second linkers can comprise a sequence with at least 90% sequence identity to SEQ ID NO: 44.

[0336] In some embodiments, the antibody comprises a CH3 domain comprising the amino acid sequence of SEQ ID NO: 141. The antibody can comprise a CH3 domain having an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 141. The antibody can comprise a CH3 domain having an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 141.

[0337] In some embodiments, the antibody comprises a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 126 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 131. The antibody can comprise a heavy chain variable region having an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 126 and a light chain variable region having an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 131. The antibody can comprise a heavy chain variable region having an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 126 and a light chain variable region having an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 131.

[0338] In some embodiments, the antibody comprises a heavy chain having a polypeptide sequence of SEQ ID NO: 135 and a light chain having the polypeptide sequenceAttorney Docket No. TP385350WO1 of SEQ ID NO: 137. The antibody can comprise a heavy chain having an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 135 and a light chain having an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 137. The antibody can comprise a heavy chain having an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 135 and a light chain having an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 137.

[0339] In some embodiments, the antibody comprises a heavy chain that is encoded by a polynucleotide sequence of SEQ ID NO: 134 and a light chain that is encoded by a polynucleotide sequence of SEQ ID NO: 136.

[0340] Disclosed herein are antibodies that immunospecifically bind to pERK1 / 2, wherein the antibodies comprise, consist essentially of, or consist of a heavy chain (HC) and a light chain (LC) that pair to form a binding domain comprising a heavy chain complementarity determining region 1 (HCDR1) comprising the amino acid sequence of SEQ ID NO: 85, a HCDR2 comprising the amino acid sequence of SEQ ID NO: 86, a HCDR3 comprising the amino acid sequence of SEQ ID NO: 87, a light chain complementarity determining region 1 (LCDR1) comprising the amino acid sequence of SEQ ID NO: 91, a LCDR2 comprising the amino acid sequence of KVS, and a LCDR3 comprising the amino acid sequence of SEQ ID NO: 92, and wherein the heavy chain further comprises a first CH2 domain, a second CH2 domain, and a third CH2 domain, wherein the first CH2 domain and the second CH2 domain are connected by a first linker and an optional first hinge, and wherein the second CH2 domain and the third CH2 domain are connected by a second linker and an optional second hinge. In some embodiments, the antibodies that immunospecifically bind to pERK1 / 2 further comprise a CH3 domain linked to the C- terminus of the third CH2 domain.

[0341] The first CH2 domain, the second CH2 domain, and the third CH2 domain can each comprise the amino acid sequence of SEQ ID NO: 104. In some embodiments, the first CH2 domain, the second CH2 domain, and the third CH2 domain each comprise the amino acid sequence of SEQ ID NO: 104. The first CH2 domain, the second CH2 domain, and the third CH2 domain can each comprise an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 104. The first CH2 domain, the second CH2 domain, and the third CH2 domain can each comprise an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 104.Attorney Docket No. TP385350WO1

[0342] The first hinge and the second hinge can each comprise the amino acid sequence of SEQ ID NO: 102. In some embodiments, the first hinge and the second hinge can each comprise the amino acid sequence of SEQ ID NO: 102. The first hinge and the second hinge can each comprise an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 102. The first hinge and the second hinge can each comprise an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 102.

[0343] The first linker and the second linker can each comprise the amino acid sequence of GS, GGS, SEQ ID NO: 14, SEQ ID NO: 38, SEQ ID NO: 43, or SEQ ID NO: 44. The first and the second linkers can comprise GS. The first and the second linkers can comprise GGS. The first and the second linkers can comprise SEQ ID NO: 14. The first and the second linkers can comprise SEQ ID NO: 38. The first and the second linkers can comprise SEQ ID NO: 43. The first and the second linkers can comprise a sequence with at least 90% sequence identity to SEQ ID NO: 43. The first and the second linkers can comprise SEQ ID NO: 44. The first and the second linkers can comprise a sequence with at least 90% sequence identity to SEQ ID NO: 44.

[0344] In some embodiments, the antibody comprises a CH3 domain comprising the amino acid sequence of SEQ ID NO: 106. The antibody can comprise a CH3 domain having an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 106. The antibody can comprise a CH3 domain having an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 106.

[0345] In some embodiments, the antibody comprises a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 84 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 90. The antibody can comprise a heavy chain variable region having an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 84 and a light chain variable region having an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 90. The antibody can comprise a heavy chain variable region having an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 84 and a light chain variable region having an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 90.

[0346] In some embodiments, the antibody comprises a heavy chain having a polypeptide sequence of SEQ ID NO: 94 and a light chain having the polypeptide sequence of SEQ ID NO: 96. The antibody can comprise a heavy chain having an amino acid sequenceAttorney Docket No. TP385350WO1 with at least 90% sequence identity to SEQ ID NO: 94 and a light chain having an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 96. The antibody can comprise a heavy chain having an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 94 and a light chain having an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 96.

[0347] In some embodiments, the antibody comprises a heavy chain that is encoded by a polynucleotide sequence of SEQ ID NO: 93 and a light chain that is encoded by a polynucleotide sequence of SEQ ID NO: 95.

[0348] Disclosed herein are antibodies that immunospecifically bind to PSD-95, wherein the antibodies comprise, consist essentially of, or consist of a heavy chain (HC) and a light chain (LC) that pair to form a binding domain comprising a heavy chain complementarity determining region 1 (HCDR1) comprising the amino acid sequence of SEQ ID NO: 111, a HCDR2 comprising the amino acid sequence of SEQ ID NO: 112, a HCDR3 comprising the amino acid sequence of SEQ ID NO: 113, a light chain complementarity determining region 1 (LCDR1) comprising the amino acid sequence of SEQ ID NO: 116, a LCDR2 comprising the amino acid sequence of DTS, and a LCDR3 comprising the amino acid sequence of SEQ ID NO: 117, and wherein the heavy chain further comprises a first CH2 domain, a second CH2 domain, and a third CH2 domain, wherein the first CH2 domain and the second CH2 domain are connected by a first linker and an optional first hinge, and wherein the second CH2 domain and the third CH2 domain are connected by a second linker and an optional second hinge. In some embodiments, the antibodies that immunospecifically bind to PSD-95 further comprise a CH3 domain linked to the C-terminus of the third CH2 domain.

[0349] The first CH2 domain, the second CH2 domain, and the third CH2 domain can each comprise the amino acid sequence of SEQ ID NO: 104. In some embodiments, the first CH2 domain, the second CH2 domain, and the third CH2 domain each comprise the amino acid sequence of SEQ ID NO: 104. The first CH2 domain, the second CH2 domain, and the third CH2 domain can each comprise an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 104. The first CH2 domain, the second CH2 domain, and the third CH2 domain can each comprise an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 104.Attorney Docket No. TP385350WO1

[0350] The first hinge and the second hinge can each comprise the amino acid sequence of SEQ ID NO: 102. In some embodiments, the first hinge and the second hinge can each comprise the amino acid sequence of SEQ ID NO: 102. The first hinge and the second hinge can each comprise an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 102. The first hinge and the second hinge can each comprise an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 102.

[0351] The first linker and the second linker can each comprise the amino acid sequence of GS, GGS, SEQ ID NO: 14, SEQ ID NO: 38, SEQ ID NO: 43, or SEQ ID NO: 44. The first and the second linkers can comprise GS. The first and the second linkers can comprise GGS. The first and the second linkers can comprise SEQ ID NO: 14. The first and the second linkers can comprise SEQ ID NO: 38. The first and the second linkers can comprise SEQ ID NO: 43. The first and the second linkers can comprise a sequence with at least 90% sequence identity to SEQ ID NO: 43. The first and the second linkers can comprise SEQ ID NO: 44. The first and the second linkers can comprise a sequence with at least 90% sequence identity to SEQ ID NO: 44.

[0352] In some embodiments, the antibody comprises a CH3 domain comprising the amino acid sequence of SEQ ID NO: 106. The antibody can comprise a CH3 domain having an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 106. The antibody can comprise a CH3 domain having an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 106.

[0353] In some embodiments, the antibody comprises a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 110 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 115. The antibody can comprise a heavy chain variable region having an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 110 and a light chain variable region having an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 115. The antibody can comprise a heavy chain variable region having an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 110 and a light chain variable region having an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 115.

[0354] In some embodiments, the antibody comprises a heavy chain having a polypeptide sequence of SEQ ID NO: 119 and a light chain having the polypeptide sequence of SEQ ID NO: 121. The antibody can comprise a heavy chain having an amino acidAttorney Docket No. TP385350WO1 sequence with at least 90% sequence identity to SEQ ID NO: 119 and a light chain having an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 121. The antibody can comprise a heavy chain having an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 119 and a light chain having an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 121.

[0355] In some embodiments, the antibody comprises a heavy chain that is encoded by a polynucleotide sequence of SEQ ID NO: 118 and a light chain that is encoded by a polynucleotide sequence of SEQ ID NO: 120.

[0356] Disclosed herein are antibodies that immunospecifically bind to TIGIT, wherein the antibodies comprise, consist essentially of, or consist of a heavy chain (HC) and a light chain (LC) that pair to form a binding domain comprising a heavy chain complementarity determining region 1 (HCDR1) comprising the amino acid sequence of SEQ ID NO: 146, a HCDR2 comprising the amino acid sequence of SEQ ID NO: 147, a HCDR3 comprising the amino acid sequence of SEQ ID NO: 148, a light chain complementarity determining region 1 (LCDR1) comprising the amino acid sequence of SEQ ID NO: 151, a LCDR2 comprising the amino acid sequence of SAS, and a LCDR3 comprising the amino acid sequence of SEQ ID NO: 152, and wherein the heavy chain further comprises a first CH2 domain, a second CH2 domain, and a third CH2 domain, wherein the first CH2 domain and the second CH2 domain are connected by a first linker and an optional first hinge, and wherein the second CH2 domain and the third CH2 domain are connected by a second linker and an optional second hinge. In some embodiments, the antibodies that immunospecifically bind to TIGIT further comprise a CH3 domain linked to the C-terminus of the third CH2 domain.

[0357] The first CH2 domain, the second CH2 domain, and the third CH2 domain can each comprise the amino acid sequence of SEQ ID NO: 19. In some embodiments, the first CH2 domain, the second CH2 domain, and the third CH2 domain each comprise the amino acid sequence of SEQ ID NO: 19. The first CH2 domain, the second CH2 domain, and the third CH2 domain can each comprise an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 19. The first CH2 domain, the second CH2 domain, and the third CH2 domain can each comprise an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 19.Attorney Docket No. TP385350WO1

[0358] The first hinge and the second hinge can each comprise the amino acid sequence of SEQ ID NO: 18. In some embodiments, the first hinge and the second hinge can each comprise the amino acid sequence of SEQ ID NO: 18. The first hinge and the second hinge can each comprise an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 18. The first hinge and the second hinge can each comprise an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 18.

[0359] The first linker and the second linker can each comprise the amino acid sequence of GS, GGS, SEQ ID NO: 14, SEQ ID NO: 38, SEQ ID NO: 43, or SEQ ID NO: 44. The first and the second linkers can comprise GS. The first and the second linkers can comprise GGS. The first and the second linkers can comprise SEQ ID NO: 14. The first and the second linkers can comprise SEQ ID NO: 38. The first and the second linkers can comprise SEQ ID NO: 43. The first and the second linkers can comprise a sequence with at least 90% sequence identity to SEQ ID NO: 43. The first and the second linkers can comprise SEQ ID NO: 44. The first and the second linkers can comprise a sequence with at least 90% sequence identity to SEQ ID NO: 44.

[0360] In some embodiments, the antibody comprises a CH3 domain comprising the amino acid sequence of SEQ ID NO: 20. The antibody can comprise a CH3 domain having an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 20. The antibody can comprise a CH3 domain having an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 20.

[0361] In some embodiments, the antibody comprises a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 145 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 150. The antibody can comprise a heavy chain variable region having an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 145 and a light chain variable region having an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 150. The antibody can comprise a heavy chain variable region having an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 145 and a light chain variable region having an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 150.

[0362] In some embodiments, the antibody comprises a heavy chain having a polypeptide sequence of SEQ ID NO: 154 and a light chain having the polypeptide sequence of SEQ ID NO: 156. The antibody can comprise a heavy chain having an amino acidAttorney Docket No. TP385350WO1 sequence with at least 90% sequence identity to SEQ ID NO: 154 and a light chain having an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 156. The antibody can comprise a heavy chain having an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 154 and a light chain having an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 156.

[0363] In some embodiments, the antibody comprises a heavy chain that is encoded by a polynucleotide sequence of SEQ ID NO: 153 and a light chain that is encoded by a polynucleotide sequence of SEQ ID NO: 155.

[0364] In some embodiments, the antibody is labeled. The label can be selected from a fluorophore, a fluorescent protein, a phosphorescent dye, a tandem dye, a polymer dye, a particle, an electron transfer agent, a hapten, an enzyme, a radioisotope, a chromophore, and an oligonucleotide-based fluorescent dye. The label can be a fluorophore. The label can be a fluorescent protein. The label can be a phosphorescent dye. The label can be a tandem dye. The label can be a polymer dye. The label can be a particle. The label can be an electron transfer agent. The label can be a hapten. The label can be an enzyme. The label can be a radioisotope. The label can be a chromophore. The label can be an oligonucleotide-based fluorescent dye.

[0365] In some embodiments, the label is a fluorophore. The fluorophore can be a pyrene, an anthracene, a naphthalene, an acridine, a stilbene, an indole or benzindole, an oxazole or benzoxazole, a thiazole or benzothiazole, an indacene, a 4-amino-7-nitrobenz-2- oxa-1,3-diazole (NBD), a cyanine, a carbocyanine, a benzocyanine, a carbostyryl, a porphyrin, a salicylate, an anthranilate, an azulene, a perylene, a pyridine, a quinoline, a borapolyazaindacene, a xanthene, a rhodamine, a silicon rhodamine, a fluorescein, a rhodol, a rosamine, an oxazine or a benzoxazine, a benzophenoxazine, a carbazine, a phenalenone, a coumarin, a furan or a benzofuran, a benzophenalenone, a benzopyrilium, a styryl, a squarine, a resorufin, an anthraquinone, a quinazolinone, an oligonucleotide-based fluorescent dyes, a conjugated polymer dye, or a tandem dye. The fluorophore can be a pyrene. The fluorophore can be an anthracene. The fluorophore can be a naphthalene. The fluorophore can be an acridine. The fluorophore can be a stilbene. The fluorophore can be an indole. The fluorophore can be a benzindole. The fluorophore can be an oxazole. The fluorophore can be a benzoxazole. The fluorophore can be a thiazole. The fluorophore can be a benzothiazole. The fluorophore can be an indacene. The fluorophore can be a 4-amino-7-nitrobenz-2-oxa-1,Attorney Docket No. TP385350WO1 3-diazole (NBD). The fluorophore can be a cyanine. The fluorophore can be a carbocyanine. The fluorophore can be a benzocyanine. The fluorophore can be a carbostyryl. The fluorophore can be a porphyrin. The fluorophore can be a salicylate. The fluorophore can be an anthranilate. The fluorophore can be an azulene. The fluorophore can be a perylene. The fluorophore can be a pyridine. The fluorophore can be a quinoline. The fluorophore can be a borapolyazaindacene. The fluorophore can be a xanthene. The fluorophore can be a rhodamine. The fluorophore can be a silicon rhodamine. The fluorophore can be a fluorescein. The fluorophore can be a rhodol. The fluorophore can be a rosamine. The fluorophore can be an oxazine. The fluorophore can be a benzoxazine. The fluorophore can be a benzophenoxazine. The fluorophore can be a carbazine. The fluorophore can be a phenalenone. The fluorophore can be a coumarin. The fluorophore can be a furan. The fluorophore can be a benzofuran. The fluorophore can be a benzophenalenone. The fluorophore can be a benzopyrilium. The fluorophore can be a styryl. The fluorophore can be a squarine. The fluorophore can be a resorufin. The fluorophore can be an anthrquinone. The fluorophore can be a quinazolinone. The fluorophore can be an oligonucleotide-based fluorescent dye. The fluorophore can be a conjugated polymer dye. The fluorophore can be a tandem dye.

[0366] In some embodiments, the label is a particle. The particle can be a semiconductor nanocrystal.

[0367] In some embodiments, the label is a hapten. The hapten can be a biotin or a biotin derivative. The hapten can be biotin. The hapten can be a biotin derivative.

[0368] In some embodiments, the label is an enzyme. The enzyme can be a peroxidase, a phosphatase, a glycosidase, and a luciferase. The enzyme can be a peroxidase. The enzyme can be a phosphatase. The enzyme can be a glycosidase. The enzyme can be a luciferase.

[0369] Disclosed herein are nucleic acid molecules comprising, consisting essentially of, or consisting of polynucleotides encoding antibodies that immunospecifically binds to Parkin, wherein the antibodies comprise a heavy chain (HC) and a light chain (LC) that pair to form a binding domain comprising a heavy chain complementarity determining region 1 (HCDR1) comprising the amino acid sequence of SEQ ID NO: 4, a HCDR2 comprising the amino acid sequence of SEQ ID NO: 5, a HCDR3 comprising the amino acid sequence of SEQ ID NO: 6, a light chain complementarity determining region 1 (LCDR1)Attorney Docket No. TP385350WO1 comprising the amino acid sequence of SEQ ID NO: 13, a LCDR2 comprising the amino acid sequence RAS, and a LCDR3 comprising the amino acid sequence of SEQ ID NO: 15, and wherein the heavy chain further comprises a first CH2 domain, a second CH2 domain, and a third CH2 domain, wherein the first CH2 domain and the second CH2 domain are connected by a first linker and an optional first hinge, and wherein the second CH2 domain and the third CH2 domain are connected by a second linker and an optional second hinge. In some embodiments, polynucleotides further encode a CH3 domain linked to the C-terminus of the third CH2 domain.

[0370] The polynucleotide can encode a first CH2 domain, a second CH2 domain, and a third CH2 domain each comprising the amino acid sequence of SEQ ID NO: 19 or SEQ ID NO: 49. In some embodiments, the polynucleotide encodes a first CH2 domain, a second CH2 domain, and a third CH2 domain each comprising the amino acid sequence of SEQ ID NO: 19. The polynucleotide can encode a first CH2 domain, a second CH2 domain, and a third CH2 domain each comprising an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 19. The polynucleotide can encode a first CH2 domain, a second CH2 domain, and a third CH2 domain each comprising an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 19. In some embodiments, in the polynucleotide encoding the first CH2 domain, the second CH2 domain, and the third CH2 domain, the polynucleotide comprises SEQ ID NO: 23.

[0371] In some embodiments, the polynucleotide encodes a first CH2 domain, a second CH2 domain, and a third CH2 domain each comprising the amino acid sequence of SEQ ID NO: 49. The polynucleotide can encode a first CH2 domain, a second CH2 domain, and a third CH2 domain each comprising an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 49. The polynucleotide can encode a first CH2 domain, a second CH2 domain, and a third CH2 domain each comprising an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 49. In some embodiments, in the polynucleotide encoding the first CH2 domain, the second CH2 domain, and the third CH2 domain, the polynucleotide encoding each CH2 domain comprises SEQ ID NO: 62.

[0372] The polynucleotide can encode a first hinge and a second hinge each comprising the amino acid sequence of SEQ ID NO: 18 or SEQ ID NO: 48. In some embodiments, the polynucleotide encodes a first hinge and a second hinge each comprising the amino acid sequence of SEQ ID NO: 18. The polynucleotide can encode a first hinge andAttorney Docket No. TP385350WO1 a second hinge each comprising an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 18. The polynucleotide can encode a first hinge and a second hinge each comprising an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 18. In some embodiments, in the polynucleotide encoding the first hinge and the second hinge, the polynucleotide encoding each hinge comprises SEQ ID NO: 22.

[0373] In some embodiments, the polynucleotide encodes a first hinge and a second hinge each comprising the amino acid sequence of SEQ ID NO: 48. The polynucleotide can encode a first hinge and a second hinge each comprising an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 48. The polynucleotide can encode a first hinge and a second hinge each comprising an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 48. In some embodiments, in the polynucleotide encoding the first hinge and the second hinge, the polynucleotide encoding each hinge comprises SEQ ID NO: 51.

[0374] The polynucleotide can encode a first linker and a second linker comprising the amino acid sequence of GS, GGS, SEQ ID NO: 14, SEQ ID NO: 38, SEQ ID NO: 43, or SEQ ID NO: 44. The polynucleotide can encode the first and the second linkers comprising GS. The first and the second linkers can comprise GGS. The polynucleotide can encode the first and the second linkers comprising SEQ ID NO: 14. The polynucleotide can encode the first and the second linkers comprising SEQ ID NO: 38. The polynucleotide can encode the first and the second linkers comprising SEQ ID NO: 43. The polynucleotide can encode a first and a second linker comprising a sequence with at least 90% sequence identity to SEQ ID NO: 43. The polynucleotide can encode the first and the second linkers comprising SEQ ID NO: 44. The polynucleotide can encode a first and a second linker comprising a sequence with at least 90% sequence identity to SEQ ID NO: 44.

[0375] In some embodiments, the polynucleotide encodes an antibody that comprises a CH3 domain comprising the amino acid sequence of SEQ ID NO: 20. In some embodiments, the polynucleotide encodes an antibody that comprises a CH3 domain having an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 20. In some embodiments, the polynucleotide encodes an antibody that comprises a CH3 domain having an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 20.

[0376] In some embodiments, the polynucleotide encodes an antibody comprising a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 7 and a lightAttorney Docket No. TP385350WO1 chain variable region having the polypeptide sequence of SEQ ID NO: 16. The polynucleotide can encode an antibody comprising a heavy chain variable region having an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 7 and a light chain variable region having an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 16. The polynucleotide can encode an antibody comprising a heavy chain variable region having an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 7 and a light chain variable region having an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 16.

[0377] In some embodiments, the polynucleotide encodes an antibody comprising a heavy chain having a polypeptide sequence of SEQ ID NO: 3 and a light chain having the polypeptide sequence of SEQ ID NO: 12. The polynucleotide can encode an antibody comprising a heavy chain having an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 3 and a light chain having an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 12. The polynucleotide can encode an antibody comprising a heavy chain having an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 3 and a light chain having an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 12. In some embodiments, in the polynucleotide encoding the heavy chain and the light chain, the polynucleotide comprises SEQ ID NO: 2 and SEQ ID NO: 11.

[0378] Disclosed herein are nucleic acid molecules comprising, consisting essentially of, or consisting of polynucleotides encoding antibodies that immunospecifically binds to OCT4, wherein the antibody comprises a heavy chain (HC) and a light chain (LC) that pair to form a binding domain comprising a heavy chain complementarity determining region 1 (HCDR1) comprising the amino acid sequence of SEQ ID NO: 29, a HCDR2 comprising the amino acid sequence of SEQ ID NO: 30, and a HCDR3 comprising the amino acid sequence of SEQ ID NO: 31, a light chain complementarity determining region 1 (LCDR1) comprising the amino acid sequence of SEQ ID NO: 37, a LCDR2 comprising the amino acid sequence GAS, and a LCDR3 comprising the amino acid sequence of SEQ ID NO: 39, and wherein the heavy chain further comprises a first CH2 domain, a second CH2 domain, and a third CH2 domain, wherein the first CH2 domain and the second CH2 domain are connected by a first linker and an optional first hinge, and wherein the second CH2 domain and the third CH2 domain are connected by a second linker and an optional secondAttorney Docket No. TP385350WO1 hinge. In some embodiments, polynucleotides further encode a CH3 domain linked to the C- terminus of the third CH2 domain.

[0379] The polynucleotide can encode a first CH2 domain, a second CH2 domain, and a third CH2 domain each comprising the amino acid sequence of SEQ ID NO: 19 or SEQ ID NO: 49. In some embodiments, the polynucleotide encodes a first CH2 domain, a second CH2 domain, and a third CH2 domain each comprising the amino acid sequence of SEQ ID NO: 19. The polynucleotide can encode a first CH2 domain, a second CH2 domain, and a third CH2 domain each comprising an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 19. The polynucleotide can encode a first CH2 domain, a second CH2 domain, and a third CH2 domain each comprising an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 19. In some embodiments, in the polynucleotide encoding the first CH2 domain, the second CH2 domain, and the third CH2 domain, the polynucleotide comprises SEQ ID NO: 23.

[0380] In some embodiments, the polynucleotide encodes a first CH2 domain, a second CH2 domain, and a third CH2 domain each comprising the amino acid sequence of SEQ ID NO: 49. The polynucleotide can encode a first CH2 domain, a second CH2 domain, and a third CH2 domain each comprising an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 49. The polynucleotide can encode a first CH2 domain, a second CH2 domain, and a third CH2 domain each comprising an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 49. In some embodiments, in the polynucleotide encoding the first CH2 domain, the second CH2 domain, and the third CH2 domain, the polynucleotide encoding each CH2 domain comprises SEQ ID NO: 62.

[0381] The polynucleotide can encode a first hinge and a second hinge each comprising the amino acid sequence of SEQ ID NO: 18 or SEQ ID NO: 48. In some embodiments, the polynucleotide encodes a first hinge and a second hinge each comprising the amino acid sequence of SEQ ID NO: 18. The polynucleotide can encode a first hinge and a second hinge each comprising an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 18. The polynucleotide can encode a first hinge and a second hinge each comprising an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 18. In some embodiments, in the polynucleotide encoding the first hinge and the second hinge, the polynucleotide encoding each hinge comprises SEQ ID NO: 22.Attorney Docket No. TP385350WO1

[0382] In some embodiments, the polynucleotide encodes a first hinge and a second hinge each comprising the amino acid sequence of SEQ ID NO: 48. The polynucleotide can encode a first hinge and a second hinge each comprising an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 48. The polynucleotide can encode a first hinge and a second hinge each comprising an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 48. In some embodiments, in the polynucleotide encoding the first hinge and the second hinge, the polynucleotide encoding each hinge comprises SEQ ID NO: 51.

[0383] The polynucleotide can encode a first linker and a second linker comprising the amino acid sequence of GS, GGS, SEQ ID NO: 14, SEQ ID NO: 38, SEQ ID NO: 43, or SEQ ID NO: 44. The polynucleotide can encode the first and the second linkers comprising GS. The first and the second linkers can comprise GGS. The polynucleotide can encode the first and the second linkers comprising SEQ ID NO: 14. The polynucleotide can encode the first and the second linkers comprising SEQ ID NO: 38. The polynucleotide can encode the first and the second linkers comprising SEQ ID NO: 43. The polynucleotide can encode a first and a second linker comprising a sequence with at least 90% sequence identity to SEQ ID NO: 43. The polynucleotide can encode the first and the second linkers comprising SEQ ID NO: 44. The polynucleotide can encode a first and a second linker comprising a sequence with at least 90% sequence identity to SEQ ID NO: 44.

[0384] In some embodiments, the polynucleotide encodes an antibody comprising a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 32 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 40. The polynucleotide can encode an antibody comprising a heavy chain variable region having an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 32 and a light chain variable region having an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 40. The polynucleotide can encode an antibody comprising a heavy chain variable region having an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 32 and a light chain variable region having an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 40.

[0385] In some embodiments, the polynucleotide encodes an antibody that comprises a CH3 domain comprising the amino acid sequence of SEQ ID NO: 20. In some embodiments, the polynucleotide encodes an antibody that comprises a CH3 domain havingAttorney Docket No. TP385350WO1 an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 20. In some embodiments, the polynucleotide encodes an antibody that comprises a CH3 domain having an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 20.

[0386] In some embodiments, the polynucleotide encodes an antibody comprising a heavy chain having a polypeptide sequence of SEQ ID NO: 28 and a light chain having the polypeptide sequence of SEQ ID NO: 36. The polynucleotide can encode an antibody comprising a heavy chain having an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 28 and a light chain having an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 36. The polynucleotide can encode an antibody comprising a heavy chain having an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 28 and a light chain having an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 36. In some embodiments, in the polynucleotide encoding the heavy chain and the light chain, the polynucleotide comprises SEQ ID NO: 27 and SEQ ID NO: 35.

[0387] Disclosed herein are nucleic acid molecules comprising, consisting essentially of, or consisting of polynucleotides encoding antibodies that immunospecifically binds to IDO1, wherein the antibodies comprise a heavy chain (HC) and a light chain (LC) that pair to form a binding domain comprising a heavy chain complementarity determining region 1 (HCDR1) comprising the amino acid sequence of SEQ ID NO: 66, a HCDR2 comprising the amino acid sequence of SEQ ID NO: 67, a HCDR3 comprising the amino acid sequence of SEQ ID NO: 68, a light chain complementarity determining region 1 (LCDR1) comprising the amino acid sequence of SEQ ID NO: 72, a LCDR2 comprising the amino acid sequence of YAS, and a LCDR3 comprising the amino acid sequence of SEQ ID NO: 73, and wherein the heavy chain further comprises a first CH2 domain, a second CH2 domain, and a third CH2 domain, wherein the first CH2 domain and the second CH2 domain are connected by a first linker and an optional first hinge, and wherein the second CH2 domain and the third CH2 domain are connected by a second linker and an optional second hinge. In some embodiments, polynucleotides further encode a CH3 domain linked to the C- terminus of the third CH2 domain.

[0388] The polynucleotide can encode a first CH2 domain, a second CH2 domain, and a third CH2 domain each comprising the amino acid sequence of SEQ ID NO: 19 or SEQ ID NO: 49. In some embodiments, the polynucleotide encodes a first CH2 domain, a second CH2 domain, and a third CH2 domain each comprising the amino acid sequence of SEQ IDAttorney Docket No. TP385350WO1 NO: 19. The polynucleotide can encode a first CH2 domain, a second CH2 domain, and a third CH2 domain each comprising an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 19. The polynucleotide can encode a first CH2 domain, a second CH2 domain, and a third CH2 domain each comprising an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 19. In some embodiments, in the polynucleotide encoding the first CH2 domain, the second CH2 domain, and the third CH2 domain, the polynucleotide comprises SEQ ID NO: 178.

[0389] In some embodiments, the polynucleotide encodes a first CH2 domain, a second CH2 domain, and a third CH2 domain each comprising the amino acid sequence of SEQ ID NO: 49. The polynucleotide can encode a first CH2 domain, a second CH2 domain, and a third CH2 domain each comprising an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 49. The polynucleotide can encode a first CH2 domain, a second CH2 domain, and a third CH2 domain each comprising an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 49. In some embodiments, in the polynucleotide encoding the first CH2 domain, the second CH2 domain, and the third CH2 domain, the polynucleotide encoding each CH2 domain comprises SEQ ID NO: 62.

[0390] The polynucleotide can encode a first hinge and a second hinge each comprising the amino acid sequence of SEQ ID NO: 18 or SEQ ID NO: 48. In some embodiments, the polynucleotide encodes a first hinge and a second hinge each comprising the amino acid sequence of SEQ ID NO: 18. The polynucleotide can encode a first hinge and a second hinge each comprising an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 18. The polynucleotide can encode a first hinge and a second hinge each comprising an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 18. In some embodiments, in the polynucleotide encoding the first hinge and the second hinge, the polynucleotide encoding each hinge comprises SEQ ID NO: 22.

[0391] In some embodiments, the polynucleotide encodes a first hinge and a second hinge each comprising the amino acid sequence of SEQ ID NO: 48. The polynucleotide can encode a first hinge and a second hinge each comprising an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 48. The polynucleotide can encode a first hinge and a second hinge each comprising an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 48. In some embodiments, in the polynucleotideAttorney Docket No. TP385350WO1 encoding the first hinge and the second hinge, the polynucleotide encoding each hinge comprises SEQ ID NO: 51.

[0392] The polynucleotide can encode a first linker and a second linker comprising the amino acid sequence of GS, GGS, SEQ ID NO: 14, SEQ ID NO: 38, SEQ ID NO: 43, or SEQ ID NO: 44. The polynucleotide can encode the first and the second linkers comprising GS. The first and the second linkers can comprise GGS. The polynucleotide can encode the first and the second linkers comprising SEQ ID NO: 14. The polynucleotide can encode the first and the second linkers comprising SEQ ID NO: 38. The polynucleotide can encode the first and the second linkers comprising SEQ ID NO: 43. The polynucleotide can encode a first and a second linker comprising a sequence with at least 90% sequence identity to SEQ ID NO: 43. The polynucleotide can encode the first and the second linkers comprising SEQ ID NO: 44. The polynucleotide can encode a first and a second linker comprising a sequence with at least 90% sequence identity to SEQ ID NO: 44.

[0393] In some embodiments, the polynucleotide encodes an antibody that comprises a CH3 domain comprising the amino acid sequence of SEQ ID NO: 20. In some embodiments, the polynucleotide encodes an antibody that comprises a CH3 domain having an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 20. In some embodiments, the polynucleotide encodes an antibody that comprises a CH3 domain having an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 20.

[0394] In some embodiments, the polynucleotide encodes an antibody comprising a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 65 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 71. The polynucleotide can encode an antibody comprising a heavy chain variable region having an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 65 and a light chain variable region having an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 71. The polynucleotide can encode an antibody comprising a heavy chain variable region having an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 65 and a light chain variable region having an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 71.

[0395] In some embodiments, the polynucleotide encodes an antibody comprising a heavy chain having a polypeptide sequence of SEQ ID NO: 75 and a light chain having the polypeptide sequence of SEQ ID NO: 77. The polynucleotide can encode an antibodyAttorney Docket No. TP385350WO1 comprising a heavy chain having an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 75 and a light chain having an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 77. The polynucleotide can encode an antibody comprising a heavy chain having an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 75 and a light chain having an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 77. In some embodiments, in the polynucleotide encoding the heavy chain and the light chain, the polynucleotide comprises SEQ ID NO: 75 and SEQ ID NO: 77.

[0396] Disclosed herein are nucleic acid molecules comprising, consisting essentially of, or consisting of polynucleotides encoding antibodies that immunospecifically binds to FLAG-tag, wherein the antibodies comprise a heavy chain (HC) and a light chain (LC) that pair to form a binding domain comprising a heavy chain complementarity determining region 1 (HCDR1) comprising the amino acid sequence of SEQ ID NO: 163, a HCDR2 comprising the amino acid sequence of SEQ ID NO: 164, a HCDR3 comprising the amino acid sequence of SEQ ID NO: 165, a light chain complementarity determining region 1 (LCDR1) comprising the amino acid sequence of SEQ ID NO: 168, a LCDR2 comprising the amino acid sequence of YAS, and a LCDR3 comprising the amino acid sequence of SEQ ID NO: 169, and wherein the heavy chain further comprises a first CH2 domain, a second CH2 domain, and a third CH2 domain, wherein the first CH2 domain and the second CH2 domain are connected by a first linker and an optional first hinge, and wherein the second CH2 domain and the third CH2 domain are connected by a second linker and an optional second hinge. In some embodiments, polynucleotides further encode a CH3 domain linked to the C-terminus of the third CH2 domain.

[0397] The polynucleotide can encode a first CH2 domain, a second CH2 domain, and a third CH2 domain each comprising the amino acid sequence of SEQ ID NO: 19 or SEQ ID NO: 49. In some embodiments, the polynucleotide encodes a first CH2 domain, a second CH2 domain, and a third CH2 domain each comprising the amino acid sequence of SEQ ID NO: 19. The polynucleotide can encode a first CH2 domain, a second CH2 domain, and a third CH2 domain each comprising an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 19. The polynucleotide can encode a first CH2 domain, a second CH2 domain, and a third CH2 domain each comprising an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 19. In some embodiments, in the polynucleotideAttorney Docket No. TP385350WO1 encoding the first CH2 domain, the second CH2 domain, and the third CH2 domain, the polynucleotide comprises SEQ ID NO: 23.

[0398] In some embodiments, the polynucleotide encodes a first CH2 domain, a second CH2 domain, and a third CH2 domain each comprising the amino acid sequence of SEQ ID NO: 49. The polynucleotide can encode a first CH2 domain, a second CH2 domain, and a third CH2 domain each comprising an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 49. The polynucleotide can encode a first CH2 domain, a second CH2 domain, and a third CH2 domain each comprising an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 49. In some embodiments, in the polynucleotide encoding the first CH2 domain, the second CH2 domain, and the third CH2 domain, the polynucleotide encoding each CH2 domain comprises SEQ ID NO: 62.

[0399] The polynucleotide can encode a first hinge and a second hinge each comprising the amino acid sequence of SEQ ID NO: 18 or SEQ ID NO: 48. In some embodiments, the polynucleotide encodes a first hinge and a second hinge each comprising the amino acid sequence of SEQ ID NO: 18. The polynucleotide can encode a first hinge and a second hinge each comprising an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 18. The polynucleotide can encode a first hinge and a second hinge each comprising an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 18. In some embodiments, in the polynucleotide encoding the first hinge and the second hinge, the polynucleotide encoding each hinge comprises SEQ ID NO: 22.

[0400] In some embodiments, the polynucleotide encodes a first hinge and a second hinge each comprising the amino acid sequence of SEQ ID NO: 48. The polynucleotide can encode a first hinge and a second hinge each comprising an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 48. The polynucleotide can encode a first hinge and a second hinge each comprising an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 48. In some embodiments, in the polynucleotide encoding the first hinge and the second hinge, the polynucleotide encoding each hinge comprises SEQ ID NO: 51.

[0401] The polynucleotide can encode a first linker and a second linker comprising the amino acid sequence of GS, GGS, SEQ ID NO: 14, SEQ ID NO: 38, SEQ ID NO: 43, or SEQ ID NO: 44. The polynucleotide can encode the first and the second linkers comprising GS. The first and the second linkers can comprise GGS. The polynucleotide can encode theAttorney Docket No. TP385350WO1 first and the second linkers comprising SEQ ID NO: 14. The polynucleotide can encode the first and the second linkers comprising SEQ ID NO: 38. The polynucleotide can encode the first and the second linkers comprising SEQ ID NO: 43. The polynucleotide can encode a first and a second linker comprising a sequence with at least 90% sequence identity to SEQ ID NO: 43. The polynucleotide can encode the first and the second linkers comprising SEQ ID NO: 44. The polynucleotide can encode a first and a second linker comprising a sequence with at least 90% sequence identity to SEQ ID NO: 44.

[0402] In some embodiments, the polynucleotide encodes an antibody that comprises a CH3 domain comprising the amino acid sequence of SEQ ID NO: 20. In some embodiments, the polynucleotide encodes an antibody that comprises a CH3 domain having an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 20. In some embodiments, the polynucleotide encodes an antibody that comprises a CH3 domain having an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 20.

[0403] In some embodiments, the polynucleotide encodes an antibody comprising a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 162 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 167. The polynucleotide can encode an antibody comprising a heavy chain variable region having an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 162 and a light chain variable region having an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 167. The polynucleotide can encode an antibody comprising a heavy chain variable region having an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 162 and a light chain variable region having an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 167.

[0404] In some embodiments, the polynucleotide encodes an antibody comprising a heavy chain having a polypeptide sequence of SEQ ID NO: 171 and a light chain having the polypeptide sequence of SEQ ID NO: 173. The polynucleotide can encode an antibody comprising a heavy chain having an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 171 and a light chain having an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 173. The polynucleotide can encode an antibody comprising a heavy chain having an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 171 and a light chain having an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 173. In some embodiments, in the polynucleotide encodingAttorney Docket No. TP385350WO1 the heavy chain and the light chain, the polynucleotide comprises SEQ ID NO: 170 and SEQ ID NO: 172.

[0405] Disclosed herein are nucleic acid molecules comprising, consisting essentially of, or consisting of polynucleotides encoding rabbit anti-mouse IgG secondary antibodies (RAM), wherein the antibodies comprise a heavy chain (HC) and a light chain (LC) that pair to form a binding domain comprising a heavy chain complementarity determining region 1 (HCDR1) comprising the amino acid sequence of SEQ ID NO: 127, a HCDR2 comprising the amino acid sequence of SEQ ID NO: 128, a HCDR3 comprising the amino acid sequence of SEQ ID NO: 129, a light chain complementarity determining region 1 (LCDR1) comprising the amino acid sequence of SEQ ID NO: 132, a LCDR2 comprising the amino acid sequence of GAS, and a LCDR3 comprising the amino acid sequence of SEQ ID NO: 133, and wherein the heavy chain further comprises a first CH2 domain, a second CH2 domain, and a third CH2 domain, wherein the first CH2 domain and the second CH2 domain are connected by a first linker and an optional first hinge, and wherein the second CH2 domain and the third CH2 domain are connected by a second linker and an optional second hinge. In some embodiments, polynucleotides further encode a CH3 domain linked to the C-terminus of the third CH2 domain.

[0406] The polynucleotide can encode a first CH2 domain, a second CH2 domain, and a third CH2 domain each comprising the amino acid sequence of SEQ ID NO: 19 or SEQ ID NO: 49. In some embodiments, the polynucleotide encodes a first CH2 domain, a second CH2 domain, and a third CH2 domain each comprising the amino acid sequence of SEQ ID NO: 19. The polynucleotide can encode a first CH2 domain, a second CH2 domain, and a third CH2 domain each comprising an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 19. The polynucleotide can encode a first CH2 domain, a second CH2 domain, and a third CH2 domain each comprising an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 19. In some embodiments, in the polynucleotide encoding the first CH2 domain, the second CH2 domain, and the third CH2 domain, the polynucleotide comprises SEQ ID NO: 23.

[0407] In some embodiments, the polynucleotide encodes a first CH2 domain, a second CH2 domain, and a third CH2 domain each comprising the amino acid sequence of SEQ ID NO: 49. The polynucleotide can encode a first CH2 domain, a second CH2 domain, and a third CH2 domain each comprising an amino acid sequence with at least 90% sequenceAttorney Docket No. TP385350WO1 identity to SEQ ID NO: 49 The polynucleotide can encode a first CH2 domain, a second CH2 domain, and a third CH2 domain each comprising an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 49. In some embodiments, in the polynucleotide encoding the first CH2 domain, the second CH2 domain, and the third CH2 domain, the polynucleotide encoding each CH2 domain comprises SEQ ID NO: 62.

[0408] The polynucleotide can encode a first hinge and a second hinge each comprising the amino acid sequence of SEQ ID NO: 18 or SEQ ID NO: 48. In some embodiments, the polynucleotide encodes a first hinge and a second hinge each comprising the amino acid sequence of SEQ ID NO: 18. The polynucleotide can encode a first hinge and a second hinge each comprising an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 18. The polynucleotide can encode a first hinge and a second hinge each comprising an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 18. In some embodiments, in the polynucleotide encoding the first hinge and the second hinge, the polynucleotide encoding each hinge comprises SEQ ID NO: 22.

[0409] In some embodiments, the polynucleotide encodes a first hinge and a second hinge each comprising the amino acid sequence of SEQ ID NO: 48. The polynucleotide can encode a first hinge and a second hinge each comprising an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 48. The polynucleotide can encode a first hinge and a second hinge each comprising an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 48. In some embodiments, in the polynucleotide encoding the first hinge and the second hinge, the polynucleotide encoding each hinge comprises SEQ ID NO: 51.

[0410] The polynucleotide can encode a first linker and a second linker comprising the amino acid sequence of GS, GGS, SEQ ID NO: 14, SEQ ID NO: 38, SEQ ID NO: 43, or SEQ ID NO: 44. The polynucleotide can encode the first and the second linkers comprising GS. The first and the second linkers can comprise GGS. The polynucleotide can encode the first and the second linkers comprising SEQ ID NO: 14. The polynucleotide can encode the first and the second linkers comprising SEQ ID NO: 38. The polynucleotide can encode the first and the second linkers comprising SEQ ID NO: 43. The polynucleotide can encode a first and a second linker comprising a sequence with at least 90% sequence identity to SEQ ID NO: 43. The polynucleotide can encode the first and the second linkers comprising SEQAttorney Docket No. TP385350WO1 ID NO: 44. The polynucleotide can encode a first and a second linker comprising a sequence with at least 90% sequence identity to SEQ ID NO: 44.

[0411] In some embodiments, the polynucleotide encodes an antibody that comprises a CH3 domain comprising the amino acid sequence of SEQ ID NO: 20. In some embodiments, the polynucleotide encodes an antibody that comprises a CH3 domain having an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 20. In some embodiments, the polynucleotide encodes an antibody that comprises a CH3 domain having an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 20.

[0412] In some embodiments, the polynucleotide encodes an antibody comprising a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 126 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 131. The polynucleotide can encode an antibody comprising a heavy chain variable region having an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 126 and a light chain variable region having an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 131. The polynucleotide can encode an antibody comprising a heavy chain variable region having an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 126 and a light chain variable region having an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 131.

[0413] In some embodiments, the polynucleotide encodes an antibody comprising a heavy chain having a polypeptide sequence of SEQ ID NO: 135 and a light chain having the polypeptide sequence of SEQ ID NO: 137. The polynucleotide can encode an antibody comprising a heavy chain having an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 135 and a light chain having an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 137. The polynucleotide can encode an antibody comprising a heavy chain having an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 135 and a light chain having an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 137. In some embodiments, in the polynucleotide encoding the heavy chain and the light chain, the polynucleotide comprises SEQ ID NO: 134 and SEQ ID NO: 136.

[0414] Disclosed herein are nucleic acid molecules comprising, consisting essentially of, or consisting of polynucleotides encoding antibodies that immunospecifically binds to pERK1 / 2, wherein the antibodies comprise a heavy chain (HC) and a light chainAttorney Docket No. TP385350WO1 (LC) that pair to form a binding domain comprising a heavy chain complementarity determining region 1 (HCDR1) comprising the amino acid sequence of SEQ ID NO: 85, a HCDR2 comprising the amino acid sequence of SEQ ID NO: 86, a HCDR3 comprising the amino acid sequence of SEQ ID NO: 87, a light chain complementarity determining region 1 (LCDR1) comprising the amino acid sequence of SEQ ID NO: 91, a LCDR2 comprising the amino acid sequence of KVS, and a LCDR3 comprising the amino acid sequence of SEQ ID NO: 92, and wherein the heavy chain further comprises a first CH2 domain, a second CH2 domain, and a third CH2 domain, wherein the first CH2 domain and the second CH2 domain are connected by a first linker and an optional first hinge, and wherein the second CH2 domain and the third CH2 domain are connected by a second linker and an optional second hinge. In some embodiments, polynucleotides further encode a CH3 domain linked to the C- terminus of the third CH2 domain.

[0415] The polynucleotide can encode a first CH2 domain, a second CH2 domain, and a third CH2 domain each comprising the amino acid sequence of SEQ ID NO: 104. In some embodiments, the polynucleotide encodes a first CH2 domain, a second CH2 domain, and a third CH2 domain each comprising the amino acid sequence of SEQ ID NO: 104. The polynucleotide can encode a first CH2 domain, a second CH2 domain, and a third CH2 domain each comprising an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 104. The polynucleotide can encode a first CH2 domain, a second CH2 domain, and a third CH2 domain each comprising an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 104. In some embodiments, in the polynucleotide encoding the first CH2 domain, the second CH2 domain, and the third CH2 domain, the polynucleotide comprises SEQ ID NO: 103.

[0416] The polynucleotide can encode a first hinge and a second hinge each comprising the amino acid sequence of SEQ ID NO: 102. In some embodiments, the polynucleotide encodes a first hinge and a second hinge each comprising the amino acid sequence of SEQ ID NO: 102. The polynucleotide can encode a first hinge and a second hinge each comprising an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 102. The polynucleotide can encode a first hinge and a second hinge each comprising an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 102. In some embodiments, in the polynucleotide encoding the first hinge and the second hinge, the polynucleotide encoding each hinge comprises SEQ ID NO: 101.Attorney Docket No. TP385350WO1

[0417] The polynucleotide can encode a first linker and a second linker comprising the amino acid sequence of GS, GGS, SEQ ID NO: 14, SEQ ID NO: 38, SEQ ID NO: 43, or SEQ ID NO: 44. The polynucleotide can encode the first and the second linkers comprising GS. The first and the second linkers can comprise GGS. The polynucleotide can encode the first and the second linkers comprising SEQ ID NO: 14. The polynucleotide can encode the first and the second linkers comprising SEQ ID NO: 38. The polynucleotide can encode the first and the second linkers comprising SEQ ID NO: 43. The polynucleotide can encode a first and a second linker comprising a sequence with at least 90% sequence identity to SEQ ID NO: 43. The polynucleotide can encode the first and the second linkers comprising SEQ ID NO: 44. The polynucleotide can encode a first and a second linker comprising a sequence with at least 90% sequence identity to SEQ ID NO: 44.

[0418] In some embodiments, the polynucleotide encodes an antibody that comprises a CH3 domain comprising the amino acid sequence of SEQ ID NO: 106. In some embodiments, the polynucleotide encodes an antibody that comprises a CH3 domain having an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 106. In some embodiments, the polynucleotide encodes an antibody that comprises a CH3 domain having an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 106.

[0419] In some embodiments, the polynucleotide encodes an antibody comprising a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 84 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 90. The polynucleotide can encode an antibody comprising a heavy chain variable region having an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 84 and a light chain variable region having an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 90. The polynucleotide can encode an antibody comprising a heavy chain variable region having an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 84 and a light chain variable region having an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 90.

[0420] In some embodiments, the polynucleotide encodes an antibody comprising a heavy chain having a polypeptide sequence of SEQ ID NO: 94 and a light chain having the polypeptide sequence of SEQ ID NO: 96. The polynucleotide can encode an antibody comprising a heavy chain having an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 94 and a light chain having an amino acid sequence with at least 90%Attorney Docket No. TP385350WO1 sequence identity to SEQ ID NO: 96. The polynucleotide can encode an antibody comprising a heavy chain having an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 94 and a light chain having an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 96. In some embodiments, in the polynucleotide encoding the heavy chain and the light chain, the polynucleotide comprises SEQ ID NO: 93 and SEQ ID NO: 95.

[0421] Disclosed herein are nucleic acid molecules comprising, consisting essentially of, or consisting of polynucleotides encoding antibodies that immunospecifically binds to PSD-95, wherein the antibodies comprise a heavy chain (HC) and a light chain (LC) that pair to form a binding domain comprising a heavy chain complementarity determining region 1 (HCDR1) comprising the amino acid sequence of SEQ ID NO: 111, a HCDR2 comprising the amino acid sequence of SEQ ID NO: 112, a HCDR3 comprising the amino acid sequence of SEQ ID NO: 113, a light chain complementarity determining region 1 (LCDR1) comprising the amino acid sequence of SEQ ID NO: 116, a LCDR2 comprising the amino acid sequence of DTS, and a LCDR3 comprising the amino acid sequence of SEQ ID NO: 117, and wherein the heavy chain further comprises a first CH2 domain, a second CH2 domain, and a third CH2 domain, wherein the first CH2 domain and the second CH2 domain are connected by a first linker and an optional first hinge, and wherein the second CH2 domain and the third CH2 domain are connected by a second linker and an optional second hinge. In some embodiments, polynucleotides further encode a CH3 domain linked to the C- terminus of the third CH2 domain.

[0422] The polynucleotide can encode a first CH2 domain, a second CH2 domain, and a third CH2 domain each comprising the amino acid sequence of SEQ ID NO: 104. In some embodiments, the polynucleotide encodes a first CH2 domain, a second CH2 domain, and a third CH2 domain each comprising the amino acid sequence of SEQ ID NO: 104. The polynucleotide can encode a first CH2 domain, a second CH2 domain, and a third CH2 domain each comprising an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 104. The polynucleotide can encode a first CH2 domain, a second CH2 domain, and a third CH2 domain each comprising an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 104. In some embodiments, in the polynucleotide encoding the first CH2 domain, the second CH2 domain, and the third CH2 domain, the polynucleotide comprises SEQ ID NO: 103.Attorney Docket No. TP385350WO1

[0423] The polynucleotide can encode a first hinge and a second hinge each comprising the amino acid sequence of SEQ ID NO: 102. In some embodiments, the polynucleotide encodes a first hinge and a second hinge each comprising the amino acid sequence of SEQ ID NO: 102. The polynucleotide can encode a first hinge and a second hinge each comprising an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 102. The polynucleotide can encode a first hinge and a second hinge each comprising an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 102. In some embodiments, in the polynucleotide encoding the first hinge and the second hinge, the polynucleotide encoding each hinge comprises SEQ ID NO: 101.

[0424] The polynucleotide can encode a first linker and a second linker comprising the amino acid sequence of GS, GGS, SEQ ID NO: 14, SEQ ID NO: 38, SEQ ID NO: 43, or SEQ ID NO: 44. The polynucleotide can encode the first and the second linkers comprising GS. The first and the second linkers can comprise GGS. The polynucleotide can encode the first and the second linkers comprising SEQ ID NO: 14. The polynucleotide can encode the first and the second linkers comprising SEQ ID NO: 38. The polynucleotide can encode the first and the second linkers comprising SEQ ID NO: 43. The polynucleotide can encode a first and a second linker comprising a sequence with at least 90% sequence identity to SEQ ID NO: 43. The polynucleotide can encode the first and the second linkers comprising SEQ ID NO: 44. The polynucleotide can encode a first and a second linker comprising a sequence with at least 90% sequence identity to SEQ ID NO: 44.

[0425] In some embodiments, the polynucleotide encodes an antibody that comprises a CH3 domain comprising the amino acid sequence of SEQ ID NO: 106. In some embodiments, the polynucleotide encodes an antibody that comprises a CH3 domain having an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 106. In some embodiments, the polynucleotide encodes an antibody that comprises a CH3 domain having an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 106.

[0426] In some embodiments, the polynucleotide encodes an antibody comprising a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 110 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 115. The polynucleotide can encode an antibody comprising a heavy chain variable region having an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 110 and a light chain variable region having an amino acid sequence with at least 90% sequence identity toAttorney Docket No. TP385350WO1 SEQ ID NO: 115. The polynucleotide can encode an antibody comprising a heavy chain variable region having an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 110 and a light chain variable region having an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 115.

[0427] In some embodiments, the polynucleotide encodes an antibody comprising a heavy chain having a polypeptide sequence of SEQ ID NO: 119 and a light chain having the polypeptide sequence of SEQ ID NO: 121. The polynucleotide can encode an antibody comprising a heavy chain having an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 119 and a light chain having an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 121. The polynucleotide can encode an antibody comprising a heavy chain having an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 119 and a light chain having an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 121. In some embodiments, in the polynucleotide encoding the heavy chain and the light chain, the polynucleotide comprises SEQ ID NO: 118 and SEQ ID NO: 120.

[0428] Disclosed herein are nucleic acid molecules comprising, consisting essentially of, or consisting of polynucleotides encoding antibodies that immunospecifically binds to TIGIT, wherein the antibodies comprise a heavy chain (HC) and a light chain (LC) that pair to form a binding domain comprising a heavy chain complementarity determining region 1 (HCDR1) comprising the amino acid sequence of SEQ ID NO: 146, a HCDR2 comprising the amino acid sequence of SEQ ID NO: 147, a HCDR3 comprising the amino acid sequence of SEQ ID NO: 148, a light chain complementarity determining region 1 (LCDR1) comprising the amino acid sequence of SEQ ID NO: 151, a LCDR2 comprising the amino acid sequence of SAS, and a LCDR3 comprising the amino acid sequence of SEQ ID NO: 152, and wherein the heavy chain further comprises a first CH2 domain, a second CH2 domain, and a third CH2 domain, wherein the first CH2 domain and the second CH2 domain are connected by a first linker and an optional first hinge, and wherein the second CH2 domain and the third CH2 domain are connected by a second linker and an optional second hinge. In some embodiments, polynucleotides further encode a CH3 domain linked to the C- terminus of the third CH2 domain.

[0429] The polynucleotide can encode a first CH2 domain, a second CH2 domain, and a third CH2 domain each comprising the amino acid sequence of SEQ ID NO: 19. InAttorney Docket No. TP385350WO1 some embodiments, the polynucleotide encodes a first CH2 domain, a second CH2 domain, and a third CH2 domain each comprising the amino acid sequence of SEQ ID NO: 19. The polynucleotide can encode a first CH2 domain, a second CH2 domain, and a third CH2 domain each comprising an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 19. The polynucleotide can encode a first CH2 domain, a second CH2 domain, and a third CH2 domain each comprising an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 19. In some embodiments, in the polynucleotide encoding the first CH2 domain, the second CH2 domain, and the third CH2 domain, the polynucleotide comprises SEQ ID NO: 23.

[0430] The polynucleotide can encode a first hinge and a second hinge each comprising the amino acid sequence of SEQ ID NO: 18. In some embodiments, the polynucleotide encodes a first hinge and a second hinge each comprising the amino acid sequence of SEQ ID NO: 18. The polynucleotide can encode a first hinge and a second hinge each comprising an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 18. The polynucleotide can encode a first hinge and a second hinge each comprising an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 18. In some embodiments, in the polynucleotide encoding the first hinge and the second hinge, the polynucleotide encoding each hinge comprises SEQ ID NO: 22.

[0431] The polynucleotide can encode a first hinge and a second hinge each comprising the amino acid sequence of SEQ ID NO: 48. In some embodiments, the polynucleotide encodes a first hinge and a second hinge each comprising the amino acid sequence of SEQ ID NO: 48. The polynucleotide can encode a first hinge and a second hinge each comprising an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 48. The polynucleotide can encode a first hinge and a second hinge each comprising an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 48. In some embodiments, in the polynucleotide encoding the first hinge and the second hinge, the polynucleotide encoding each hinge comprises SEQ ID NO: 51.

[0432] The polynucleotide can encode a first linker and a second linker comprising the amino acid sequence of GS, GGS, SEQ ID NO: 14, SEQ ID NO: 38, SEQ ID NO: 43, or SEQ ID NO: 44. The polynucleotide can encode the first and the second linkers comprising GS. The first and the second linkers can comprise GGS. The polynucleotide can encode the first and the second linkers comprising SEQ ID NO: 14. The polynucleotide can encode theAttorney Docket No. TP385350WO1 first and the second linkers comprising SEQ ID NO: 38. The polynucleotide can encode the first and the second linkers comprising SEQ ID NO: 43. The polynucleotide can encode a first and a second linker comprising a sequence with at least 90% sequence identity to SEQ ID NO: 43. The polynucleotide can encode the first and the second linkers comprising SEQ ID NO: 44. The polynucleotide can encode a first and a second linker comprising a sequence with at least 90% sequence identity to SEQ ID NO: 44.

[0433] In some embodiments, the polynucleotide encodes an antibody that comprises a CH3 domain comprising the amino acid sequence of SEQ ID NO: 20. In some embodiments, the polynucleotide encodes an antibody that comprises a CH3 domain having an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 20. In some embodiments, the polynucleotide encodes an antibody that comprises a CH3 domain having an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 20.

[0434] In some embodiments, the polynucleotide encodes an antibody comprising a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 145 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 150. The polynucleotide can encode an antibody comprising a heavy chain variable region having an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 145 and a light chain variable region having an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 150. The polynucleotide can encode an antibody comprising a heavy chain variable region having an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 145 and a light chain variable region having an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 150.

[0435] In some embodiments, the polynucleotide encodes an antibody comprising a heavy chain having a polypeptide sequence of SEQ ID NO: 154 and a light chain having the polypeptide sequence of SEQ ID NO: 156. The polynucleotide can encode an antibody comprising a heavy chain having an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 154 and a light chain having an amino acid sequence with at least 90% sequence identity to SEQ ID NO: 156. The polynucleotide can encode an antibody comprising a heavy chain having an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 154 and a light chain having an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 156. In some embodiments, in the polynucleotide encodingAttorney Docket No. TP385350WO1 the heavy chain and the light chain, the polynucleotide comprises SEQ ID NO: 153 and SEQ ID NO: 155.

[0436] Disclosed herein are vectors comprising any of the herein described nucleic acid molecules that comprise polynucleotides encoding antibodies. Disclosed herein are also host cells comprising any of the herein disclosed vectors. Suitable host cells include eukaryotic cells and prokaryotic cells.

[0437] An antibody or polypeptide according to the present disclosure may be produced using any known and well-established expression system and recombinant cell culturing technology, for example, by expression in bacterial hosts (prokaryotic systems) or eukaryotic systems such as yeasts, fungi, insect cells or mammalian cells. An antibody or polypeptide of the present disclosure may be produced in transgenic organisms such as a goat or a plant. An antibody or polypeptide of the present disclosure may also be produced by chemical synthesis.

[0438] Disclosed herein are methods of detecting the presence of a target protein in a sample, the methods comprising, consisting essentially of, or consisting of applying any of the herein disclosed isolated polypeptides or any of the herein disclosed antibodies to a sample containing a target protein, incubating the isolated polypeptide or the antibody in the sample under conditions permitting the isolated polypeptide or the antibody to bind to the target protein and to form a bound isolated polypeptide or a bound antibody, and determining the presence of the bound isolated polypeptide or the bound antibody, thereby detecting the presence of the target protein.

[0439] Disclosed herein are methods of detecting the presence of a target protein in a sample, the methods comprising, consisting essentially of, or consisting of applying any of the herein disclosed isolated polypeptides to a sample containing a target protein, incubating the isolated polypeptide in the sample under conditions permitting the isolated polypeptide to bind to the target protein and to form a bound isolated polypeptide, and determining the presence of the bound isolated polypeptide, thereby detecting the presence of the target protein.

[0440] Disclosed herein are methods of detecting the presence of a target protein in a sample, the methods comprising, consisting essentially of, or consisting of applying any of the herein disclosed antibodies to a sample containing a target protein, incubating the antibody in the sample under conditions permitting the antibody to bind to the target proteinAttorney Docket No. TP385350WO1 and to form a bound antibody, and determining the presence of the bound antibody, thereby detecting the presence of the target protein.

[0441] Disclosed herein are uses of any of the herein disclosed isolated polypeptides or any of the herein disclosed antibodies in detecting the presence of a target protein in a sample. The uses can comprise, consist essentially of, or consist of applying any of the herein disclosed isolated polypeptides or any of the herein disclosed antibodies to a sample containing a target protein, incubating the isolated polypeptide or the antibody in the sample under conditions permitting the isolated polypeptide or the antibody to bind to the target protein and to form a bound isolated polypeptide or a bound antibody, and determining the presence of the bound isolated polypeptide or the bound antibody, thereby detecting the presence of the target protein.

[0442] Disclosed herein are uses of any of the herein disclosed isolated polypeptides in detecting the presence of a target protein in a sample. The uses can comprise, consist essentially of, or consist of applying any of the herein disclosed isolated polypeptides to a sample containing a target protein, incubating the isolated polypeptide in the sample under conditions permitting the isolated polypeptide to bind to the target protein and to form a bound isolated polypeptide, and determining the presence of the bound isolated polypeptide, thereby detecting the presence of the target protein.

[0443] Disclosed herein are uses of any of the herein disclosed antibodies in detecting the presence of a target protein in a sample. The uses can comprise, consist essentially of, or consist of applying any of the herein disclosed antibodies to a sample containing a target protein, incubating the antibody in the sample under conditions permitting the antibody to bind to the target protein and to form a bound antibody, and determining the presence of the bound antibody, thereby detecting the presence of the target protein.

[0444] In some embodiments of the methods or uses, a lesser amount of herein disclosed isolated polypeptide or antibody is required relative to an amount of a wild type isolated polypeptide or wild type antibody that would be required for generating a detectable signal in an equivalent method. A lesser amount of a herein disclosed isolated polypeptide can be required relative to an amount of a wild type isolated peptide that would be required for generating a detectable signal in an equivalent method. A lesser amount of a herein disclosed antibody can be required relative to an amount of a wild type antibody that would be required for generating a detectable signal in an equivalent method. The herein disclosedAttorney Docket No. TP385350WO1 isolated polypeptides and antibodies provide for enhanced sensitivity at lower concentrations which allows for robust signal to be generated at concentrations at which a wild type isolated polypeptide or wild type antibody would provide less signal.

[0445] In some embodiments of the methods or uses, a lesser quantity of sample is used (e.g., rare clinical samples) but greater signal is still generated relative to an equivalent method that uses a wild type isolated polypeptide or wild type antibody.

[0446] In some embodiments of the methods or uses, a lower concentration of target protein in the sample is able to be detected relative to an equivalent method that uses a wild type isolated polypeptide or wild type antibody.

[0447] In some embodiments of the methods or uses, the herein disclosed isolated polypeptide or antibody has less background binding to cells that express Fc gamma receptors (FcγR) relative to a wild type isolated polypeptide or a wild type antibody.

[0448] In some embodiments of the methods or uses, the target protein is OCT4, Parkin, IDO1, FLAG-tag, pERK1 / 2, PSD-95, or TIGIT. In some embodiments of the methods or uses, the target protein is OCT4. In some embodiments of the methods or uses, the target protein is Parkin. In some embodiments of the methods or uses, the target protein is IDO1. In some embodiments of the methods or uses, the target protein is FLAG-tag. In some embodiments of the methods or uses, the target protein is pERK1 / 2. In some embodiments of the methods or uses, the target protein is PSD-95. In some embodiments of the methods or uses, the target is TIGIT.

[0449] The target protein can be present in the sample in low abundance. Low abundance targets are expressed or present at low levels that fall below the limit of detection or that provide a very weak signal using a wild type antibody in standard immunoapplications and immunoassays. Low abundance includes situations where there is an inability or a difficulty in detecting the target protein in a particular assay using a wild type antibody.

[0450] In some embodiments of the methods or uses, the presence of the bound isolated polypeptide or the bound antibody is determined by applying a secondary antibody to said sample, wherein the secondary antibody is labeled. In some embodiments of the methods of uses, the secondary antibody is the rabbit anti-mouse (RAM) of the present disclosure.

[0451] In some embodiments of the methods or uses, the antibody is labeled. The label can be selected from a fluorophore, a fluorescent protein, a phosphorescent dye, aAttorney Docket No. TP385350WO1 tandem dye, a polymer dye, a particle, an electron transfer agent, a hapten, an enzyme, a radioisotope, a chromophore, and an oligonucleotide-based fluorescent dye. The label can be a fluorophore. The label can be a fluorescent protein. The label can be a phosphorescent dye. The label can be a tandem dye. The label can be a polymer dye. The label can be a particle. The label can be an electron transfer agent. The label can be a hapten. The label can be an enzyme. The label can be a radioisotope. The label can be a chromophore. The label can be an oligonucleotide-based fluorescent dye.

[0452] In some embodiments, the label is a fluorophore. The fluorophore can be a pyrene, an anthracene, a naphthalene, an acridine, a stilbene, an indole or benzindole, an oxazole or benzoxazole, a thiazole or benzothiazole, an indacene, a 4-amino-7-nitrobenz-2- oxa-1, 3-diazole (NBD), a cyanine, a carbocyanine, a benzocyanine, a carbostyryl, a porphyrin, a salicylate, an anthranilate, an azulene, a perylene, a pyridine, a quinoline, a borapolyazaindacene, a xanthene, a rhodamine, a silicon rhodamine, a fluorescein, a rhodol, a rosamine, an oxazine or a benzoxazine, a benzophenoxazine, a carbazine, a phenalenone, a coumarin, a furan or a benzofuran, a benzophenalenone, a benzopyrilium, a styryl, a squarine, a resorufin, an anthraquinone, a quinazolinone, an oligonucleotide-based fluorescent dyes, a conjugated polymer dye, or a tandem dye. The fluorophore can be a pyrene. The fluorophore can be an anthracene. The fluorophore can be a naphthalene. The fluorophore can be an acridine. The fluorophore can be a stilbene. The fluorophore can be an indole. The fluorophore can be a benzindole. The fluorophore can be an oxazole. The fluorophore can be a benzoxazole. The fluorophore can be a thiazole. The fluorophore can be a benzothiazole. The fluorophore can be an indacene. The fluorophore can be a 4-amino-7-nitrobenz-2-oxa-1, 3-diazole (NBD). The fluorophore can be a cyanine. The fluorophore can be a carbocyanine. The fluorophore can be a benzocyanine. The fluorophore can be a carbostyryl. The fluorophore can be a porphyrin. The fluorophore can be a salicylate. The fluorophore can be an anthranilate. The fluorophore can be an azulene. The fluorophore can be a perylene. The fluorophore can be a pyridine. The fluorophore can be a quinoline. The fluorophore can be a borapolyazaindacene. The fluorophore can be a xanthene. The fluorophore can be a rhodamine. The fluorophore can be a silicon rhodamine. The fluorophore can be a fluorescein. The fluorophore can be a rhodol. The fluorophore can be a rosamine. The fluorophore can be an oxazine. The fluorophore can be a benzoxazine. The fluorophore can be a benzophenoxazine. The fluorophore can be a carbazine. The fluorophore can be aAttorney Docket No. TP385350WO1 phenalenone. The fluorophore can be a coumarin. The fluorophore can be a benzofuran. The fluorophore can be a benzophenalenone. The fluorophore can be a benzopyrilium. The fluorophore can be a styryl. The fluorophore can be a squarine. The fluorophore can be a resorufin. The fluorophore can be an anthrquinone. The fluorophore can be a quinazolinone. The fluorophore can be an oligonucleotide-based fluorescent dye. The fluorophore can be a conjugated polymer dye. The fluorophore can be a tandem dye.

[0453] In some embodiments, the label is a particle. The particle can be a semiconductor nanocrystal.

[0454] In some embodiments, the label is a hapten. The hapten can be a biotin or a biotin derivative. The hapten can be biotin. The hapten can be a biotin derivative.

[0455] In some embodiments, the label is an enzyme. The enzyme can be a peroxidase, a phosphatase, a glycosidase, and a luciferase. The enzyme can be a peroxidase. The enzyme can be a phosphatase. The enzyme can be a glycosidase. The enzyme can be a luciferase.

[0456] The methods or uses can further comprise adding an enzymatic substrate selected from a chromogenic, a fluorogenic, and a chemiluminescent substrate. The enzymatic substate can be a chromogenic substrate. The enzymatic substate can be a fluorogenic substrate. The enzymatic substate can be a chemiluminescent substrate.

[0457] The methods or uses can comprise detecting the labeled antibody utilizing flow cytometry, immunohistochemistry, immunocytochemistry, or western blotting. The labeled antibody can be detected utilizing flow cytometry. The labeled antibody can be detected utilizing immunohistochemistry. The labeled antibody can be detected utilizing immunocytochemistry. The labeled antibody can be detected utilizing western blotting. Sequences Table 1: Anti-Parkin Antibody Variable Domains Anti-Parkin Antibody Clone Name: 21H24L9 Species: Rabbit Isotype: IgG DNA Leader sequence, Heavy atggagactgggctgcgctggcttctcctggtcgctgtgctcaaaggtgtccagtgt Chain (SEQ ID NO: 1)Attorney Docket No. TP385350WO1 Heavy Chain DNA Sequence cagtcggtggaggagtccgggggtcgcctggtcacgcctgggacacccctgacact cacttgcacagtctctggattctccctcagtagctactacatgacctgggtccgccagg ctccagggaaggggctggaatacatcggatacattgcttataatggtaacacatggta cgcgagctgggcgaaaggccgattcaccatctccaaaacctcgaccacggtggatc tcaaaatcaccagtccgacaaccgaagacacggccacctatttctgtgccagagatc ggggtgatactagtggtatcgactttaacatctggggcccaggcaccctggtcaccgt ctcgagcgggcaacctaaggctccatcagttttcccactggccccctgctgcgggga cacacccagctccacggtgaccctgggctgcctggtcaaaggctacctcccggagc cagtgaccgtgacctggaactcgggcaccctcaccaatggggtacgcaccttcccgt ccgtccggcagtcctcaggcctctactcgctgagcagcgtggtgagcgtgacctcaa gcagccagcccgtcacttgcaacgtggcccacccagccaccaacaccaaagtgga caagaccgtggcgccctcgacatgcagcaagcccacgtgcccaccccctgagttcg aggggggaccgtctgtcttcatcttccccccaaaacccaaggacaccctcatgatctc acgcacccccgaggtgacctgcgtggtggtggacgtgagccaggatgaccccgag gtgcagttcacatggtacataaacaacgagcaggtccgcaccgcccggccgccgct acgggagcagcagttcaacagcacgatccgcgtggtcagcaccctccccatcgcgc accaggactggctgaggggcaaggagttcaagtgcaaagtccacaacaaggcact cccggcctccatcgagaagaccatcagcaaggccagagggggcggcggcggca gcggcggcggcggcagcctagcgccctcgacatgcagcaagcccacgtgcccac cccctgagttcgaggggggaccgtctgtcttcatcttccccccaaaacccaaggaca ccctcatgatctcacgcacccccgaggtgacctgcgtggtggtggacgtgagccag gatgaccccgaggtgcagttcacatggtacataaacaacgagcaggtccgcaccgc ccggccgccgctacgggagcagcagttcaacagcacgatccgcgtggtcagcacc ctccccatcgcgcaccaggactggctgaggggcaaggagttcaagtgcaaagtcca caacaaggcactcccggcctccatcgagaagaccatcagcaaggccagaggggg cggcggcggcagcggcggcggcggcagcctagcgccctcgacatgcagcaagc ccacgtgcccaccccctgagttcgaggggggaccgtctgtcttcatcttccccccaaa acccaaggacaccctcatgatctcacgcacccccgaggtgacctgcgtggtggtgg acgtgagccaggatgaccccgaggtgcagttcacatggtacataaacaacgagcag gtccgcaccgcccggccgccgctacgggagcagcagttcaacagcacgatccgcg tggtcagcaccctccccatcgcgcaccaggactggctgaggggcaaggagttcaag tgcaaagtccacaacaaggcactcccggcctccatcgagaagaccatcagcaaggcAttorney Docket No. TP385350WO1 cagagggcagcccctggagccgaaggtctacaccatgggtcctccccgggaggag ctgagcagcaggtcggtcagcctggcctgcatgatcaacggcttctacccttccgaca tctcggtggagtgggagaagaacgggaaggcagaggacaactacaagaccacgc cggccgtgctggacagcgacggctcctacttcctctacagcaagctctcagtgccca cgagtgagtggcagcggggcgacgtcttcacttgctccgtgatgcacgaggccttgc acaaccactacacgcagaagtccatctcccgctctccgggtaaa (SEQ ID NO: 2) Heavy Chain Amino Acid QSVEESGGRLVTPGTPLTLTCTVSGFSLSSYYMTWVR Sequence QAPGKGLEYIGYIAYNGNTWYASWAKGRFTISKTSTT VDLKITSPTTEDTATYFCARDRGDTSGIDFNIWGPGTL VTVSSGQPKAPSVFPLAPCCGDTPSSTVTLGCLVKGYL PEPVTVTWNSGTLTNGVRTFPSVRQSSGLYSLSSVVSV TSSSQPVTCNVAHPATNTKVDKTVAPSTCSKPTCPPPE FEGGPSVFIFPPKPKDTLMISRTPEVTCVVVDVSQDDP EVQFTWYINNEQVRTARPPLREQQFNSTIRVVSTLPIA HQDWLRGKEFKCKVHNKALPASIEKTISKARGGGGG SGGGGSLAPSTCSKPTCPPPEFEGGPSVFIFPPKPKDTL MISRTPEVTCVVVDVSQDDPEVQFTWYINNEQVRTAR PPLREQQFNSTIRVVSTLPIAHQDWLRGKEFKCKVHN KALPASIEKTISKARGGGGGSGGGGSLAPSTCSKPTCP PPEFEGGPSVFIFPPKPKDTLMISRTPEVTCVVVDVSQD DPEVQFTWYINNEQVRTARPPLREQQFNSTIRVVSTLP IAHQDWLRGKEFKCKVHNKALPASIEKTISKARGQPL EPKVYTMGPPREELSSRSVSLACMINGFYPSDISVEWE KNGKAEDNYKTTPAVLDSDGSYFLYSKLSVPTSEWQ RGDVFTCSVMHEALHNHYTQKSISRSPGK (SEQ ID NO: 3) Heavy Chain CDR1 Amino Acid GFSLSSYY (SEQ ID NO: 4) Sequence (IMGT) Heavy Chain CDR2 Amino Acid IAYNGNT (SEQ ID NO: 5) SequenceAttorney Docket No. TP385350WO1 (IMGT) Heavy Chain CDR3 Amino Acid ARDRGDTSGIDFNI (SEQ ID NO: 6) Sequence (IMGT) Heavy Chain Variable Region QSVEESGGRLVTPGTPLTLTCTVSGFSLSSYYMTWVR Amino Acid Sequence QAPGKGLEYIGYIAYNGNTWYASWAKGRFTISKTSTT VDLKITSPTTEDTATYFCARDRGDTSGIDFNIWGPGTL VTVSS (SEQ ID NO: 7) DNA Leader sequence, Heavy atggagactgggctgcgctggcttctcctggtcgctgtgctcaaaggtgtccagtgt Chain (SEQ ID NO: 1) Heavy Chain Fab DNA sequence cagtcggtggaggagtccgggggtcgcctggtcacgcctgggacacccctgacact cacttgcacagtctctggattctccctcagtagctactacatgacctgggtccgccagg ctccagggaaggggctggaatacatcggatacattgcttataatggtaacacatggta cgcgagctgggcgaaaggccgattcaccatctccaaaacctcgaccacggtggatc tcaaaatcaccagtccgacaaccgaagacacggccacctatttctgtgccagagatc ggggtgatactagtggtatcgactttaacatctggggcccaggcaccctggtcaccgt ctcgagcgggcaacctaaggctccatcagttttcccactggccccctgctgcgggga cacacccagctccacggtgaccctgggctgcctggtcaaaggctacctcccggagc cagtgaccgtgacctggaactcgggcaccctcaccaatggggtacgcaccttcccgt ccgtccggcagtcctcaggcctctactcgctgagcagcgtggtgagcgtgacctcaa gcagccagcccgtcacttgcaacgtggcccacccagccaccaacaccaaagtgga caagaccgtg (SEQ ID NO: 8) Heavy Chain Fab Amino Acid QSVEESGGRLVTPGTPLTLTCTVSGFSLSSYYMTWVR Sequence QAPGKGLEYIGYIAYNGNTWYASWAKGRFTISKTSTT VDLKITSPTTEDTATYFCARDRGDTSGIDFNIWGPGTL VTVSSGQPKAPSVFPLAPCCGDTPSSTVTLGCLVKGYL PEPVTVTWNSGTLTNGVRTFPSVRQSSGLYSLSSVVSV TSSSQPVTCNVAHPATNTKVDKTV (SEQ ID NO: 9) DNA Leader Sequence, Light atggacacgagggcccccactcagctgctggggctcctgctgctctggctcccaggt Chain gccacattt (SEQ ID NO: 10) Light Chain DNA Sequence gcctatgatatgacccagactccagcctctgtggaggtagctgtgggaggcacagtc accatcaagtgccaggccagtgagagcattgatacctacttagcctggtatcagcagaAttorney Docket No. TP385350WO1 aaccagggcagcctcccaagcaactgatctacagggcatccactctggcatctggg gtctcatcgcggttcaaaggcagtggagctgggacacagttcactctcaccatcagc ggcgtggagtgtgccgatgctgccacttactactgtcaacaggctactagtagtagta atattgataatgctttcggcggagggaccgaggtggtggtcaaaggtgatccagttgc acctactgtcctcatcttcccaccagctgctgatcaggtggcaactggaacagtcacc atcgtgtgtgtggcgaataaatactttcccgatgtcaccgtcacctgggaggtggatgg caccacccaaacaactggcatcgagaacagtaaaacaccgcagaattctgcagattg tacctacaacctcagcagcactctgacactgaccagcacacagtacaacagccacaa agagtacacttgcaaggtgacccagggcacgacctcagtcgtccagagcttcaatag gggtgactgt (SEQ ID NO: 11) Light Chain Amino Acid AYDMTQTPASVEVAVGGTVTIKCQASESIDTYLAWY Sequence QQKPGQPPKQLIYRASTLASGVSSRFKGSGAGTQFTLT ISGVECADAATYYCQQATSSSNIDNAFGGGTEVVVKG DPVAPTVLIFPPAADQVATGTVTIVCVANKYFPDVTV TWEVDGTTQTTGIENSKTPQNSADCTYNLSSTLTLTST QYNSHKEYTCKVTQGTTSVVQSFNRGDC (SEQ ID NO: 12) Light Chain CDR1 Amino Acid ESIDTY (SEQ ID NO: 13) Sequence (IMGT) Light Chain CDR2 Amino Acid RAS Sequence (IMGT) Light Chain CDR3 Amino Acid QQATSSSNIDNA (SEQ ID NO: 15) Sequence (IMGT) Light Chain Variable Region AYDMTQTPASVEVAVGGTVTIKCQASESIDTYLAWY Amino Acid Sequence QQKPGQPPKQLIYRASTLASGVSSRFKGSGAGTQFTLT ISGVECADAATYYCQQATSSSNIDNAFGGGTEVVVKG (SEQ ID NO: 16) DNA Leader Sequence, Light atggacacgagggcccccactcagctgctggggctcctgctgctctggctcccaggt Chain gccacattt (SEQ ID NO: 10)Attorney Docket No. TP385350WO1 Light Chain Full Length DNA gcctatgatatgacccagactccagcctctgtggaggtagctgtgggaggcacagtc sequence accatcaagtgccaggccagtgagagcattgatacctacttagcctggtatcagcaga aaccagggcagcctcccaagcaactgatctacagggcatccactctggcatctggg gtctcatcgcggttcaaaggcagtggagctgggacacagttcactctcaccatcagc ggcgtggagtgtgccgatgctgccacttactactgtcaacaggctactagtagtagta atattgataatgctttcggcggagggaccgaggtggtggtcaaaggtgatccagttgc acctactgtcctcatcttcccaccagctgctgatcaggtggcaactggaacagtcacc atcgtgtgtgtggcgaataaatactttcccgatgtcaccgtcacctgggaggtggatgg caccacccaaacaactggcatcgagaacagtaaaacaccgcagaattctgcagattg tacctacaacctcagcagcactctgacactgaccagcacacagtacaacagccacaa agagtacacttgcaaggtgacccagggcacgacctcagtcgtccagagcttcaatag gggtgactgt (SEQ ID NO: 11) Light Chain Full Length Amino AYDMTQTPASVEVAVGGTVTIKCQASESIDTYLAWY Acid sequence QQKPGQPPKQLIYRASTLASGVSSRFKGSGAGTQFTLT ISGVECADAATYYCQQATSSSNIDNAFGGGTEVVVKG DPVAPTVLIFPPAADQVATGTVTIVCVANKYFPDVTV TWEVDGTTQTTGIENSKTPQNSADCTYNLSSTLTLTST QYNSHKEYTCKVTQGTTSVVQSFNRGDC (SEQ ID NO: 12) Table 2: Anti-Parkin Parental Antibody Constant Region Anti- Parkin Parental Antibody Heavy Chain Constant Region Amino Acid Sequence (Underlined residues are mutations in LFLEPS variants) CH1 Domain Amino Acid GQPKAPSVFPLAPCCGDTPSSTVTLGCLVKGYLPEPVT Sequence VTWNSGTLTNGVRTFPSVRQSSGLYSLSSVVSVTSSSQ PVTCNVAHPATNTKVDKTV (SEQ ID NO: 17) Hinge Amino Acid Sequence APSTCSKPTCPPPELLGGP (SEQ ID NO: 18) CH2 Domain Amino Acid SVFIFPPKPKDTLMISRTPEVTCVVVDVSQDDPEVQFT sequence WYINNEQVRTARPPLREQQFNSTIRVVSTLPIAHQDW LRGKEFKCKVHNKALPAPIEKTISKARG (SEQ ID NO: 19)Attorney Docket No. TP385350WO1 CH3 Domain Amino Acid QPLEPKVYTMGPPREELSSRSVSLACMINGFYPSDISV Sequence EWEKNGKAEDNYKTTPAVLDSDGSYFLYSKLSVPTSE WQRGDVFTCSVMHEALHNHYTQKSISRSPGK (SEQ ID NO: 20) CH1 Domain DNA Sequence gggcaacctaaggctccatcagttttcccactggccccctgctgcggggacacaccc agctccacggtgaccctgggctgcctggtcaaaggctacctcccggagccagtgac cgtgacctggaactcgggcaccctcaccaatggggtacgcaccttcccgtccgtccg gcagtcctcaggcctctactcgctgagcagcgtggtgagcgtgacctcaagcagcca gcccgtcacttgcaacgtggcccacccagccaccaacaccaaagtggacaagacc gtg (SEQ ID NO: 21) Hinge DNA Sequence gcgccctcgacatgcagcaagcccacgtgcccaccccctgagctcctggggggac cg (SEQ ID NO: 22) CH2 Domain DNA sequence tctgtcttcatcttccccccaaaacccaaggacaccctcatgatctcacgcacccccga ggtgacctgcgtggtggtggacgtgagccaggatgaccccgaggtgcagttcacat ggtacataaacaacgagcaggtccgcaccgcccggccgccgctacgggagcagc agttcaacagcacgatccgcgtggtcagcaccctccccatcgcgcaccaggactgg ctgaggggcaaggagttcaagtgcaaagtccacaacaaggcactcccggcccccat cgagaagaccatcagcaaggccagaggg (SEQ ID NO: 23) CH3 Domain DNA Sequence cagcccctggagccgaaggtctacaccatgggtcctccccgggaggagctgagca gcaggtcggtcagcctggcctgcatgatcaacggcttctacccttccgacatctcggt ggagtgggagaagaacgggaaggcagaggacaactacaagaccacgccggccg tgctggacagcgacggctcctacttcctctacagcaagctctcagtgcccacgagtga gtggcagcggggcgacgtcttcacttgctccgtgatgcacgaggccttgcacaacca ctacacgcagaagtccatctcccgctctccgggtaaa (SEQ ID NO: 24) Anti- Parkin Parental Antibody DPVAPTVLIFPPAADQVATGTVTIVCVANKYFPDVTV Light Chain Constant Region TWEVDGTTQTTGIENSKTPQNSADCTYNLSSTLTLTST Amino Acid Sequence QYNSHKEYTCKVTQGTTSVVQSFNRGDC (SEQ ID NO: 25) Anti-Parkin Parental Antibody gatccagttgcacctactgtcctcatcttcccaccagctgctgatcaggtggcaactgg Light Chain Constant Region aacagtcaccatcgtgtgtgtggcgaataaatactttcccgatgtcaccgtcacctggg DNA Sequence aggtggatggcaccacccaaacaactggcatcgagaacagtaaaacaccgcagaat tctgcagattgtacctacaacctcagcagcactctgacactgaccagcacacagtacaAttorney Docket No. TP385350WO1 acagccacaaagagtacacttgcaaggtgacccagggcacgacctcagtcgtccag agcttcaataggggtgactgt (SEQ ID NO: 26) Table 3: Anti-OCT4 Antibody Variable Domains Anti-OCT4 Antibody Clone Name: 3H8L6 Species: Rabbit Isotype: IgG DNA Leader sequence, Heavy atggagactgggctgcgctggcttctcctggtcgctgtgctcaaaggtgtccagtgt Chain (SEQ ID NO: 1) Heavy Chain DNA Sequence cagtcggtggaggagtccgggggtcgcctggtcacgcctgggacacccctgacact cacttgcacagtctctggattctccctcagtaactatgcaatgagctgggtccgccagg ctccagggaaggggctggaatggatcggaatcgttagtattagtgctggcacatacta cgcgagctgggcgaaaggccgattcaccatctccaaaacctcgaccacggtggttct gagagtcaccagtccgacaaccgaggacacggccacctatttctgtgccacagggg gttatactggtaacgtctggggcccaggcaccctggtcaccgtctcgagcgggcaac ctaaggctccatcagtcttcccactggccccctgctgcggggacacacccagctcca cggtgaccctgggctgcctggtcaaaggctacctcccggagccagtgaccgtgacc tggaactcgggcaccctcaccaatggggtacgcaccttcccgtccgtccggcagtcc tcaggcctctactcgctgagcagcgtggtgagcgtgacctcaagcagccagcccgtc acttgcaacgtggcccacccagccaccaacaccaaagtggacaagaccgtggcgc cctcgacatgcagcaagcccacgtgcccaccccctgagttcgaggggggaccgtct gtcttcatcttccccccaaaacccaaggacaccctcatgatctcacgcacccccgagg tgacctgcgtggtggtggacgtgagccaggatgaccccgaggtgcagttcacatggt acataaacaacgagcaggtccgcaccgcccggccgccgctacgggagcagcagtt caacagcacgatccgcgtggtcagcaccctccccatcgcgcaccaggactggctga ggggcaaggagttcaagtgcaaagtccacaacaaggcactcccggcctccatcgag aagaccatcagcaaggccagagggggcggcggcggcagcggcggcggcggca gcctagcgccctcgacatgcagcaagcccacgtgcccaccccctgagttcgagggg ggaccgtctgtcttcatcttccccccaaaacccaaggacaccctcatgatctcacgca cccccgaggtgacctgcgtggtggtggacgtgagccaggatgaccccgaggtgcaAttorney Docket No. TP385350WO1 gttcacatggtacataaacaacgagcaggtccgcaccgcccggccgccgctacggg agcagcagttcaacagcacgatccgcgtggtcagcaccctccccatcgcgcaccag gactggctgaggggcaaggagttcaagtgcaaagtccacaacaaggcactcccgg cctccatcgagaagaccatcagcaaggccagagggggcggcggcggcagcggc ggcggcggcagcctagcgccctcgacatgcagcaagcccacgtgcccaccccctg agttcgaggggggaccgtctgtcttcatcttccccccaaaacccaaggacaccctcat gatctcacgcacccccgaggtgacctgcgtggtggtggacgtgagccaggatgacc ccgaggtgcagttcacatggtacataaacaacgagcaggtccgcaccgcccggccg ccgctacgggagcagcagttcaacagcacgatccgcgtggtcagcaccctccccat cgcgcaccaggactggctgaggggcaaggagttcaagtgcaaagtccacaacaag gcactcccggcctccatcgagaagaccatcagcaaggccagagggcagcccctgg agccgaaggtctacaccatgggtcctccccgggaggagctgagcagcaggtcggt cagcctggcctgcatgatcaacggcttctacccttccgacatctcggtggagtgggag aagaacgggaaggcagaggacaactacaagaccacgccggccgtgctggacagc gacggctcctacttcctctacagcaagctctcagtgcccacgagtgagtggcagcgg ggcgacgtcttcacttgctccgtgatgcacgaggccttgcacaaccactacacgcag aagtccatctcccgctctccgggtaaa (SEQ ID NO: 27) Heavy Chain Amino Acid QSVEESGGRLVTPGTPLTLTCTVSGFSLSNYAMSWVR Sequence QAPGKGLEWIGIVSISAGTYYASWAKGRFTISKTSTTV VLRVTSPTTEDTATYFCATGGYTGNVWGPGTLVTVSS GQPKAPSVFPLAPCCGDTPSSTVTLGCLVKGYLPEPVT VTWNSGTLTNGVRTFPSVRQSSGLYSLSSVVSVTSSSQ PVTCNVAHPATNTKVDKTVAPSTCSKPTCPPPEFEGGP SVFIFPPKPKDTLMISRTPEVTCVVVDVSQDDPEVQFT WYINNEQVRTARPPLREQQFNSTIRVVSTLPIAHQDW LRGKEFKCKVHNKALPASIEKTISKARGGGGGSGGGG SLAPSTCSKPTCPPPEFEGGPSVFIFPPKPKDTLMISRTP EVTCVVVDVSQDDPEVQFTWYINNEQVRTARPPLRE QQFNSTIRVVSTLPIAHQDWLRGKEFKCKVHNKALPA SIEKTISKARGGGGGSGGGGSLAPSTCSKPTCPPPEFEG GPSVFIFPPKPKDTLMISRTPEVTCVVVDVSQDDPEVQ FTWYINNEQVRTARPPLREQQFNSTIRVVSTLPIAHQDAttorney Docket No. TP385350WO1 WLRGKEFKCKVHNKALPASIEKTISKARGQPLEPKVY TMGPPREELSSRSVSLACMINGFYPSDISVEWEKNGK AEDNYKTTPAVLDSDGSYFLYSKLSVPTSEWQRGDVF TCSVMHEALHNHYTQKSISRSPGK (SEQ ID NO: 28) Heavy Chain CDR1 Amino Acid GFSLSNYA (SEQ ID NO: 29) Sequence (IMGT) Heavy Chain CDR2 Amino Acid VSISAGT (SEQ ID NO: 30) Sequence (IMGT) Heavy Chain CDR3 Amino Acid ATGGYTGNV (SEQ ID NO: 31) Sequence (IMGT) Heavy Chain Variable Region QSVEESGGRLVTPGTPLTLTCTVSGFSLSNYAMSWVR Amino Acid Sequence QAPGKGLEWIGIVSISAGTYYASWAKGRFTISKTSTTV VLRVTSPTTEDTATYFCATGGYTGNVWGPGTLVTVSS (SEQ ID NO: 32) DNA Leader Sequence, Heavy atggagactgggctgcgctggcttctcctggtcgctgtgctcaaaggtgtccagtgt Chain (SEQ ID NO: 1) Heavy Chain Fab DNA sequence cagtcggtggaggagtccgggggtcgcctggtcacgcctgggacacccctgacact cacttgcacagtctctggattctccctcagtaactatgcaatgagctgggtccgccagg ctccagggaaggggctggaatggatcggaatcgttagtattagtgctggcacatacta cgcgagctgggcgaaaggccgattcaccatctccaaaacctcgaccacggtggttct gagagtcaccagtccgacaaccgaggacacggccacctatttctgtgccacagggg gttatactggtaacgtctggggcccaggcaccctggtcaccgtctcgagcgggcaac ctaaggctccatcagtcttcccactggccccctgctgcggggacacacccagctcca cggtgaccctgggctgcctggtcaaaggctacctcccggagccagtgaccgtgacc tggaactcgggcaccctcaccaatggggtacgcaccttcccgtccgtccggcagtcc tcaggcctctactcgctgagcagcgtggtgagcgtgacctcaagcagccagcccgtc acttgcaacgtggcccacccagccaccaacaccaaagtggacaagaccgtg (SEQ ID NO: 33) Heavy Chain Fab Amino Acid QSVEESGGRLVTPGTPLTLTCTVSGFSLSNYAMSWVR Sequence QAPGKGLEWIGIVSISAGTYYASWAKGRFTISKTSTTV VLRVTSPTTEDTATYFCATGGYTGNVWGPGTLVTVSS GQPKAPSVFPLAPCCGDTPSSTVTLGCLVKGYLPEPVTAttorney Docket No. TP385350WO1 VTWNSGTLTNGVRTFPSVRQSSGLYSLSSVVSVTSSSQ PVTCNVAHPATNTKVDKTV (SEQ ID NO: 34) DNA Leader Sequence, Light atggacacgagggcccccactcagctgctggggctcctgctgctctggctcccaggt Chain gccacattt (SEQ ID NO: 10) Light Chain DNA Sequence gcgcaagttctgacccagactgcatcgtccgtgtctgcagctgtgggaggcacagtc accattagttgccagtccagtcagagtgtttataataagaattggttaggctggtatcag cagaaaccagggcagcctcccaggctcctgatctatggtgcatccactctggcatctg gggtcccatcgcggttcagcggcagtggatctgggacacagttcactctcaccatca gcgacgtgcagtgtgacgatgctgccacttactactgtgcaggcggttatagtaataat attgttagtttcggcggaggaaccagggtggtcgtcgaagttgctacattggcacctac tgtcctcatcttcccaccatctcctgctgagctggcaactggaacagccaccatcgtgt gcgtggcaaacaaatactttcccgatggcaccgtcacctggcaggtcgatggcaagc ccctaacaactggcatcgagaccagtaaaacaccgcagaattctgatgattgtaccta caacctcagcagtactctgacactgaaaagcgacgagtacaacagccacgacgagt acacttgccaggtggcccagggctcaggctcaccggtcgtccagagcttcaatagg ggtgactgt (SEQ ID NO: 35) Light Chain Amino Acid AQVLTQTASSVSAAVGGTVTISCQSSQSVYNKNWLG Sequence WYQQKPGQPPRLLIYGASTLASGVPSRFSGSGSGTQFT LTISDVQCDDAATYYCAGGYSNNIVSFGGGTRVVVEV ATLAPTVLIFPPSPAELATGTATIVCVANKYFPDGTVT WQVDGKPLTTGIETSKTPQNSDDCTYNLSSTLTLKSD EYNSHDEYTCQVAQGSGSPVVQSFNRGDC (SEQ ID NO: 36) Light Chain CDR1 Amino Acid QSVYNKNW (SEQ ID NO: 37) Sequence (IMGT) Light Chain CDR2 Amino Acid GAS Sequence (IMGT) Light Chain CDR3 Amino Acid AGGYSNNIVS (SEQ ID NO: 39) Sequence (IMGT) Light Chain Variable Region AQVLTQTASSVSAAVGGTVTISCQSSQSVYNKNWLG Amino Acid Sequence WYQQKPGQPPRLLIYGASTLASGVPSRFSGSGSGTQFTAttorney Docket No. TP385350WO1 LTISDVQCDDAATYYCAGGYSNNIVSFGGGTRVVVE (SEQ ID NO: 40) DNA Leader sequence, Light atggacacgagggcccccactcagctgctggggctcctgctgctctggctcccaggt Chain gccacattt (SEQ ID NO: 10) Light Chain DNA sequence gcgcaagttctgacccagactgcatcgtccgtgtctgcagctgtgggaggcacagtc accattagttgccagtccagtcagagtgtttataataagaattggttaggctggtatcag cagaaaccagggcagcctcccaggctcctgatctatggtgcatccactctggcatctg gggtcccatcgcggttcagcggcagtggatctgggacacagttcactctcaccatca gcgacgtgcagtgtgacgatgctgccacttactactgtgcaggcggttatagtaataat attgttagtttcggcggaggaaccagggtggtcgtcgaagttgctacattggcacctac tgtcctcatcttcccaccatctcctgctgagctggcaactggaacagccaccatcgtgt gcgtggcaaacaaatactttcccgatggcaccgtcacctggcaggtcgatggcaagc ccctaacaactggcatcgagaccagtaaaacaccgcagaattctgatgattgtaccta caacctcagcagtactctgacactgaaaagcgacgagtacaacagccacgacgagt acacttgccaggtggcccagggctcaggctcaccggtcgtccagagcttcaatagg ggtgactgt (SEQ ID NO: 35) Light Chain Amino Acid AQVLTQTASSVSAAVGGTVTISCQSSQSVYNKNWLG sequence WYQQKPGQPPRLLIYGASTLASGVPSRFSGSGSGTQFT LTISDVQCDDAATYYCAGGYSNNIVSFGGGTRVVVEV ATLAPTVLIFPPSPAELATGTATIVCVANKYFPDGTVT WQVDGKPLTTGIETSKTPQNSDDCTYNLSSTLTLKSD EYNSHDEYTCQVAQGSGSPVVQSFNRGDC (SEQ ID NO: 36) Table 4: Anti- OCT4 Parental Antibody Constant Region Anti- OCT4 Parental Antibody Heavy Chain Constant Region Amino Acid Sequence (Underlined residues are mutations in LFLEPS variants) CH1 Domain Amino Acid GQPKAPSVFPLAPCCGDTPSSTVTLGCLVKGYLPEPVT Sequence VTWNSGTLTNGVRTFPSVRQSSGLYSLSSVVSVTSSSQ PVTCNVAHPATNTKVDKTV (SEQ ID NO: 17) Hinge Amino Acid Sequence APSTCSKPTCPPPELLGGP (SEQ ID NO: 18)Attorney Docket No. TP385350WO1 CH2 domain Amino Acid SVFIFPPKPKDTLMISRTPEVTCVVVDVSQDDPEVQFT Sequence WYINNEQVRTARPPLREQQFNSTIRVVSTLPIAHQDW LRGKEFKCKVHNKALPAPIEKTISKARG (SEQ ID NO: 19) CH3 Domain Amino Acid QPLEPKVYTMGPPREELSSRSVSLACMINGFYPSDISV Sequence EWEKNGKAEDNYKTTPAVLDSDGSYFLYSKLSVPTSE WQRGDVFTCSVMHEALHNHYTQKSISRSPGK (SEQ ID NO: 20) CH1 Domain DNA Sequence gggcaacctaaggctccatcagtcttcccactggccccctgctgcggggacacaccc agctccacggtgaccctgggctgcctggtcaaaggctacctcccggagccagtgac cgtgacctggaactcgggcaccctcaccaatggggtacgcaccttcccgtccgtccg gcagtcctcaggcctctactcgctgagcagcgtggtgagcgtgacctcaagcagcca gcccgtcacttgcaacgtggcccacccagccaccaacaccaaagtggacaagacc gtg (SEQ ID NO: 176) Hinge DNA Sequence gcgccctcgacatgcagcaagcccacgtgcccaccccctgagctcctggggggac cg (SEQ ID NO: 22) CH2 Domain DNA sequence tctgtcttcatcttccccccaaaacccaaggacaccctcatgatctcacgcacccccga ggtgacctgcgtggtggtggacgtgagccaggatgaccccgaggtgcagttcacat ggtacataaacaacgagcaggtccgcaccgcccggccgccgctacgggagcagc agttcaacagcacgatccgcgtggtcagcaccctccccatcgcgcaccaggactgg ctgaggggcaaggagttcaagtgcaaagtccacaacaaggcactcccggcccccat cgagaagaccatcagcaaggccagaggg (SEQ ID NO: 23) CH3 Domain DNA Sequence cagcccctggagccgaaggtctacaccatgggtcctccccgggaggagctgagca gcaggtcggtcagcctggcctgcatgatcaacggcttctacccttccgacatctcggt ggagtgggagaagaacgggaaggcagaggacaactacaagaccacgccggccg tgctggacagcgacggctcctacttcctctacagcaagctctcagtgcccacgagtga gtggcagcggggcgacgtcttcacttgctccgtgatgcacgaggccttgcacaacca ctacacgcagaagtccatctcccgctctccgggtaaa (SEQ ID NO: 24) Anti- OCT4 Parental Antibody VATLAPTVLIFPPSPAELATGTATIVCVANKYFPDGTV Light Chain Constant Region TWQVDGKPLTTGIETSKTPQNSDDCTYNLSSTLTLKS Amino Acid Sequence DEYNSHDEYTCQVAQGSGSPVVQSFNRGDC (SEQ ID NO: 41)Attorney Docket No. TP385350WO1 Anti- OCT4 Parental Antibody gttgctacattggcacctactgtcctcatcttcccaccatctcctgctgagctggcaact Light Chain Constant Region ggaacagccaccatcgtgtgcgtggcaaacaaatactttcccgatggcaccgtcacc DNA Sequence tggcaggtcgatggcaagcccctaacaactggcatcgagaccagtaaaacaccgca gaattctgatgattgtacctacaacctcagcagtactctgacactgaaaagcgacgagt acaacagccacgacgagtacacttgccaggtggcccagggctcaggctcaccggtc gtccagagcttcaataggggtgactgt (SEQ ID NO: 42) Table 5: Rabbit 2TdCH2 Domain Amino Acid Sequence (Underlined residues are mutations in LFLEPS variants) Hinge 1 Amino Acid Sequence APSTCSKPTCPPPELLGGP (SEQ ID NO: 18) First CH2 Domain Amino Acid SVFIFPPKPKDTLMISRTPEVTCVVVDVSQDDPEVQFT Sequence WYINNEQVRTARPPLREQQFNSTIRVVSTLPIAHQDW LRGKEFKCKVHNKALPAPIEKTISKARG (SEQ ID NO: 19) Linker 1 Amino Acid Sequence GGGGSGGGGSL (SEQ ID NO: 44) Hinge 2 Amino Acid Sequence APSTCSKPTCPPPELLGGP (SEQ ID NO: 18) Second CH2 Domain Amino SVFIFPPKPKDTLMISRTPEVTCVVVDVSQDDPEVQFT Acid Sequence WYINNEQVRTARPPLREQQFNSTIRVVSTLPIAHQDW LRGKEFKCKVHNKALPAPIEKTISKARG (SEQ ID NO: 19) Linker 2 Amino Acid Sequence GGGGSGGGGSL (SEQ ID NO: 44) Hinge 3 Amino Acid Sequence APSTCSKPTCPPPELLGGP (SEQ ID NO: 18) Third CH2 domain Amino Acid SVFIFPPKPKDTLMISRTPEVTCVVVDVSQDDPEVQFT Sequence WYINNEQVRTARPPLREQQFNSTIRVVSTLPIAHQDW LRGKEFKCKVHNKALPAPIEKTISKARG (SEQ ID NO: 19) Fc Domain (only CH2+CH3) SVFIFPPKPKDTLMISRTPEVTCVVVDVSQDDPEVQFT Amino Acid Sequence WYINNEQVRTARPPLREQQFNSTIRVVSTLPIAHQDW LRGKEFKCKVHNKALPAPIEKTISKARGQPLEPKVYT MGPPREELSSRSVSLACMINGFYPSDISVEWEKNGKAAttorney Docket No. TP385350WO1 EDNYKTTPAVLDSDGSYFLYSKLSVPTSEWQRGDVFT CSVMHEALHNHYTQKSISRSPGK (SEQ ID NO: 45) 2TdCH2 Fc Domain APSTCSKPTCPPPELLGGPSVFIFPPKPKDTLMISRTPEV TCVVVDVSQDDPEVQFTW...

Claims

Attorney Docket No. TP385350WO1 What is claimed:

1. An isolated polypeptide comprising the amino acid sequence of Formula I: Z-(X-Yn- Zr)p-X, wherein Z is a hinge; X is a CH2 domain; Y is a linker; n is an integer of 1 or greater; p is an integer of 2 or greater; and r is an integer of 0 or 1.

2. The isolated polypeptide of claim 1, wherein Z comprises an amino acid sequence comprising at least 90% identity to SEQ ID NO: 18, SEQ ID NO: 48, or SEQ ID NO: 102; X comprises an amino acid sequence comprising at least 90% identity to SEQ ID NO: 19, SEQ ID NO: 49, or SEQ ID NO: 104; Y is a linker; n is an integer of 1 or greater; p is an integer of 2 or greater; and r is an integer of 0 or 1.

3. The isolated polypeptide of claim 1 or claim 2, wherein Y comprises an amino acid sequence of GS, GGS, at least 90% identity to SEQ ID NO: 14, at least 90% identity to SEQ ID NO: 38, at least 90% identity to SEQ ID NO: 43, or at least 90% identity to SEQ ID NO:

44.

4. The isolated polypeptide of any one of claims 1 to 3, wherein Z comprises SEQ ID NO: 18, SEQ ID NO: 48, or SEQ ID NO: 102; X comprises SEQ ID NO: 19, SEQ ID NO: 49, or SEQ ID NO: 104; Y comprises GS, GGS, SEQ ID NO: 14, SEQ ID NO: 38, SEQ ID NO: 43, or SEQ ID NO: 44; n is an integer of 1 or greater; p is an integer of 2 or greater; and r is an integer of 0 or 1.Attorney Docket No. TP385350WO1 5. The isolated polypeptide of any one of claims 1 to 4, wherein Z comprises the amino acid sequence of SEQ ID NO: 18, X comprises the amino acid sequence of SEQ ID NO: 19, Y comprises the amino acid sequence of SEQ ID NO: 44, n is 1, and p is 2.

6. The isolated polypeptide of any one of claims 1 to 4, wherein Z comprises the amino acid sequence of SEQ ID NO: 48, X comprises the amino acid sequence of SEQ ID NO: 49, Y comprises the amino acid sequence of SEQ ID NO: 44, n is 1, and p is 2.

7. The isolated polypeptide of any one of claims 1 to 4, wherein Z comprises the amino acid sequence of SEQ ID NO: 102, X comprises the amino acid sequence of SEQ ID NO: 104, Y comprises the amino acid sequence of SEQ ID NO: 44, n is 1, and p is 2.

8. The isolated polypeptide of any one of the preceding claims, wherein the isolated polypeptide is linked to a second polypeptide by one or more disulfide bonds, wherein the isolated polypeptide and the second polypeptide share the same amino acid sequence.

9. The isolated polypeptide of any one of the preceding claims, wherein the isolated polypeptide comprises the amino acid sequence of SEQ ID NO: 53 or SEQ ID NO:

55.

10. The isolated polypeptide of claim 8, wherein the one or more disulfide bonds occur between the hinge of the isolated polypeptide and the hinge of the second polypeptide.

11. The isolated polypeptide of any one of the preceding claims, wherein the isolated polypeptide further comprises a CH3 domain.

12. The isolated polypeptide of claim 9, wherein the CH3 domain comprises the amino acid sequence of SEQ ID NO: 20, or SEQ ID NO:

106.

13. An antibody comprising the isolated polypeptide of any one of the preceding claims linked to a Fab domain.

14. The antibody of claim 13, wherein the Fab comprises a CH1 domain.

15. The antibody of claim 14, wherein the CH1 domain comprises the amino acid sequence of SEQ ID NO: 17, SEQ ID NO: 100, or SEQ ID NO: 160.Attorney Docket No. TP385350WO1 16. The antibody of claim 13, wherein the Fab comprises a Fab heavy chain and a Fab light chain that pair to form a binding domain.

17. The antibody of any one of claims 13 to 16, wherein the Fab immunospecifically binds to Parkin, OCT4, IDO1, FLAG-tag, pERK1 / 2, PSD-95, or TIGIT.

18. The antibody of any one of claims 13 to 17, wherein the isolated polypeptide is an antibody with the isotype IgG.

19. An antibody comprising a polypeptide comprising the amino acid sequence of Formula I: Z-(X-Yn-Zr)p-X, wherein Z is a hinge; X is a CH2 domain; Y is a linker; n is an integer of 1 or greater; p is an integer of 2 or greater; and r is an integer of 0 or 1.

20. The antibody of claim 19, wherein Z comprises an amino acid sequence comprising at least 90% identity to SEQ ID NO: 18, SEQ ID NO: 48, or SEQ ID NO: 102; X comprises an amino acid sequence comprising at least 90% identity to SEQ ID NO: 19, SEQ ID NO: 49, or SEQ ID NO: 104; Y is a linker; n is an integer of 1 or greater; p is an integer of 2 or greater; and r is an integer of 0 or 1.

21. The antibody of claim 19 or claim 20, wherein Y comprises an amino acid sequence of GS, GGS, at least 90% identity to SEQ ID NO: 14, at least 90% identity to SEQ ID NO: 38, at least 90% identity to SEQ ID NO: 43, or at least 90% identity to SEQ ID NO:

44.

22. The antibody of any one of claims 19 to 21, wherein Z comprises SEQ ID NO: 18, SEQ ID NO: 48, or SEQ ID NO: 102; X comprises SEQ ID NO: 19, SEQ ID NO: 49,Attorney Docket No. TP385350WO1 or SEQ ID NO: 104; Y comprises GS, GGS, SEQ ID NO: 14, SEQ ID NO: 38, SEQ ID NO: 43, or SEQ ID NO: 44; n is an integer of 1 or greater; p is an integer of 2 or greater; and r is an integer of 0 or 1.

23. The antibody of any one of claims 19 to 22, wherein Z comprises the amino acid sequence of SEQ ID NO: 18, X comprises the amino acid sequence of SEQ ID NO: 19, Y comprises the amino acid sequence of SEQ ID NO: 44, n is 1, and p is 2.

24. The antibody of any one of claims 19 to 22, wherein Z comprises the amino acid sequence of SEQ ID NO: 48, X comprises the amino acid sequence of SEQ ID NO: 49, Y comprises the amino acid sequence of SEQ ID NO: 44, n is 1, and p is 2.

25. The antibody of any one of claims 19 to 22, wherein Z comprises the amino acid sequence of SEQ ID NO: 102, X comprises the amino acid sequence of SEQ ID NO: 104, Y comprises the amino acid sequence of SEQ ID NO: 44, n is 1, and p is 2.

26. The antibody of any one of claims 19 to 25, wherein the polypeptide is linked to a second polypeptide by one or more disulfide bonds, wherein the polypeptide and the second polypeptide share the same amino acid sequence.

27. The antibody of any one of claims 19 to 26, wherein the polypeptide comprises the amino acid sequence of SEQ ID NO: 53 or SEQ ID NO:

55.

28. The antibody of claim 26, wherein the one or more disulfide bonds occur between the hinge of the polypeptide and the hinge of the second polypeptide.

29. The antibody of any one of claims 19 to 28, further comprising a CH3 domain.

30. The antibody of claim 29, wherein the CH3 domain comprises the amino acid sequence of SEQ ID NO: 20, or SEQ ID NO:

106.

31. The antibody of any one of claims 13 to 30, wherein the antibody has a reduction of background signal of at least 1.5-fold compared to a corresponding wildtype antibody without the isolated polypeptide.Attorney Docket No. TP385350WO1 32. The antibody of any one of claims 13 to 31, wherein the antibody has an increase in sensitivity of at least 1.5-fold compared to a corresponding wildtype antibody without the isolated polypeptide.

33. The isolated polypeptide of any one of claims 1-12 or the antibody of any one of claims 13-32, further comprising a label.

34. A nucleic acid encoding the isolated polypeptide of any one of claims 1-12 or the antibody of any one of claims 13-32.

35. An antibody that immunospecifically binds to Parkin, wherein the antibody comprises a heavy chain (HC) and a light chain (LC) that pair to form a binding domain comprising a heavy chain complementarity determining region 1 (HCDR1) comprising the amino acid sequence of SEQ ID NO: 4, a HCDR2 comprising the amino acid sequence of SEQ ID NO: 5, a HCDR3 comprising the amino acid sequence of SEQ ID NO: 6, a light chain complementarity determining region 1 (LCDR1) comprising the amino acid sequence of SEQ ID NO: 13, a LCDR2 comprising the amino acid sequence RAS, and a LCDR3 comprising the amino acid sequence of SEQ ID NO: 15, and wherein the heavy chain further comprises a first CH2 domain, a second CH2 domain, and a third CH2 domain, wherein the first CH2 domain and the second CH2 domain are connected by a first linker and an optional first hinge, and wherein the second CH2 domain and the third CH2 domain are connected by a second linker and an optional second hinge, and wherein the heavy chain further comprises a CH3 domain.

36. The antibody of claim 35, wherein the first CH2 domain, the second CH2 domain, and the third CH2 domain each comprises the amino acid sequence of SEQ ID NO: 19 or SEQ ID NO:

49.

37. The antibody of claim 35 or claim 36, wherein the first hinge and the second hinge each comprises the amino acid sequence of SEQ ID NO: 18 or SEQ ID NO: 48.Attorney Docket No. TP385350WO1 38. The antibody of any one of claims 35 to 37, wherein the first linker and the second linker each comprises the amino acid sequence of GS, GGS, SEQ ID NO: 14, SEQ ID NO: 38, SEQ ID NO: 43, or SEQ ID NO:

44.

39. The antibody of any one of claims 35 to 38, wherein the CH3 domain comprises the amino acid sequence of SEQ ID NO:

20.

40. The antibody of any one of claims 35 to 39, wherein the antibody comprises a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 7 and a light chain variable region having the polypeptide sequence of SEQ ID NO:

16.

41. The antibody of any one of claims 35 to 40, wherein the antibody comprises a heavy chain having a polypeptide sequence of SEQ ID NO: 3 and a light chain having the polypeptide sequence of SEQ ID NO:

12.

42. The antibody of any one of claims 35 to 41, wherein the antibody comprises a heavy chain that is encoded by a polynucleotide sequence of SEQ ID NO: 2 and a light chain that is encoded by a polynucleotide sequence of SEQ ID NO:

11.

43. An antibody that immunospecifically binds to OCT4, wherein the antibody comprises a heavy chain (HC) and a light chain (LC) that pair to form a binding domain comprising a heavy chain complementarity determining region 1 (HCDR1) comprising the amino acid sequence of SEQ ID NO: 29, a HCDR2 comprising the amino acid sequence of SEQ ID NO: 30, and a HCDR3 comprising the amino acid sequence of SEQ ID NO: 31, a light chain complementarity determining region 1 (LCDR1) comprising the amino acid sequence of SEQ ID NO: 37, a LCDR2 comprising the amino acid sequence GAS, and a LCDR3 comprising the amino acid sequence of SEQ ID NO: 39, and wherein the heavy chain further comprises a first CH2 domain, a second CH2 domain, and a third CH2 domain, wherein the first CH2 domain and the second CH2 domain are connected by a first linker and an optional first hinge, and wherein the second CH2 domain and the third CH2 domain are connected by a second linker and an optional second hinge, and wherein the heavy chain further comprises a CH3 domain.Attorney Docket No. TP385350WO1 44. The antibody of claim 43, wherein the first CH2 domain, the second CH2 domain, and the third CH2 domain each comprises the amino acid sequence of SEQ ID NO: 19 or SEQ ID NO:

49.

45. The antibody of claim 43 or claim 44, wherein the first hinge and the second hinge each comprises the amino acid sequence of SEQ ID NO: 18 or SEQ ID NO:

48.

46. The antibody of any one of claims 43 to 45, wherein the first linker and the second linker each comprises the amino acid sequence of GS, GGS, SEQ ID NO: 14, SEQ ID NO: 38, SEQ ID NO: 43, or SEQ ID NO:

44.

47. The antibody of any one of claims 43 to 46, wherein the CH3 domain comprises the amino acid sequence of SEQ ID NO:

20.

48. The antibody of any one of claims 43 to 47, wherein the antibody comprises a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 32 and a light chain variable region having the polypeptide sequence of SEQ ID NO:

40.

49. The antibody of any one of claims 43 to 48, wherein the antibody comprises a heavy chain having a polypeptide sequence of SEQ ID NO: 28 and a light chain having the polypeptide sequence of SEQ ID NO:

36.

50. The antibody of any one of claims 43 to 49, wherein the antibody comprises a heavy chain that is encoded by a polynucleotide sequence of SEQ ID NO: 27 and a light chain that is encoded by a polynucleotide sequence of SEQ ID NO:

35.

51. An antibody that immunospecifically binds to IDO1, wherein the antibody comprises a heavy chain (HC) and a light chain (LC) that pair to form a binding domain comprising a heavy chain complementarity determining region 1 (HCDR1) comprising the amino acid sequence of SEQ ID NO: 66, a HCDR2 comprising the amino acid sequence of SEQ ID NO: 67, a HCDR3 comprising the amino acid sequence of SEQ ID NO: 68, a light chain complementarity determining region 1 (LCDR1) comprising the amino acid sequence of SEQ ID NO: 72, a LCDR2Attorney Docket No. TP385350WO1 comprising the amino acid sequence of YAS, and a LCDR3 comprising the amino acid sequence of SEQ ID NO: 73, and wherein the heavy chain further comprises a first CH2 domain, a second CH2 domain, and a third CH2 domain, wherein the first CH2 domain and the second CH2 domain are connected by a first linker and an optional first hinge, and wherein the second CH2 domain and the third CH2 domain are connected by a second linker and an optional second hinge, and wherein the heavy chain further comprises a CH3 domain.

52. The antibody of claim 51, wherein the first CH2 domain, the second CH2 domain, and the third CH2 domain each comprises the amino acid sequence of SEQ ID NO: 19 or SEQ ID NO:

49.

53. The antibody of claim 51 or claim 52, wherein the first hinge and the second hinge each comprises the amino acid sequence of SEQ ID NO: 18 or SEQ ID NO:

48.

54. The antibody of any one of claims 51 to 53, wherein the first linker and the second linker each comprises the amino acid sequence of GS, GGS, SEQ ID NO: 14, SEQ ID NO: 38, SEQ ID NO: 43, or SEQ ID NO:

44.

55. The antibody of any one of claims 51 to 54, wherein the CH3 domain comprises the amino acid sequence of SEQ ID NO:

20.

56. The antibody of any one of claims 51 to 55, wherein the antibody comprises a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 65 and a light chain variable region having the polypeptide sequence of SEQ ID NO:

71.

57. The antibody of any one of claims 51 to 56, wherein the antibody comprises a heavy chain having a polypeptide sequence of SEQ ID NO: 75 and a light chain having the polypeptide sequence of SEQ ID NO:

77.

58. The antibody of any one of claims 51 to 57, wherein the antibody comprises a heavy chain that is encoded by a polynucleotide sequence of SEQ ID NO: 74 and a light chain that is encoded by a polynucleotide sequence of SEQ ID NO: 76.Attorney Docket No. TP385350WO1 59. An antibody that immunospecifically binds to FLAG-tag, wherein the antibody comprises a heavy chain (HC) and a light chain (LC) that pair to form a binding domain comprising a heavy chain complementarity determining region 1 (HCDR1) comprising the amino acid sequence of SEQ ID NO: 163, a HCDR2 comprising the amino acid sequence of SEQ ID NO: 164, a HCDR3 comprising the amino acid sequence of SEQ ID NO: 165, a light chain complementarity determining region 1 (LCDR1) comprising the amino acid sequence of SEQ ID NO: 168, a LCDR2 comprising the amino acid sequence of YAS, and a LCDR3 comprising the amino acid sequence of SEQ ID NO: 169, and wherein the heavy chain further comprises a first CH2 domain, a second CH2 domain, and a third CH2 domain, wherein the first CH2 domain and the second CH2 domain are connected by a first linker and an optional first hinge, and wherein the second CH2 domain and the third CH2 domain are connected by a second linker and an optional second hinge, and wherein the heavy chain further comprises a CH3 domain.

60. The antibody of claim 59, wherein the first CH2 domain, the second CH2 domain, and the third CH2 domain each comprises the amino acid sequence of SEQ ID NO: 19 or SEQ ID NO:

49.

61. The antibody of claim 59 or claim 60, wherein the first hinge and the second hinge each comprises the amino acid sequence of SEQ ID NO: 18 or SEQ ID NO:

48.

62. The antibody of any one of claims 59 to 61, wherein the first linker and the second linker each comprises the amino acid sequence of GS, GGS, SEQ ID NO: 14, SEQ ID NO: 38, SEQ ID NO: 43, or SEQ ID NO:

44.

63. The antibody of any one of claims 59 to 62, wherein the CH3 domain comprises the amino acid sequence of SEQ ID NO:

20.

64. The antibody of any one of claims 59 to 63, wherein the antibody comprises a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 162 and a light chain variable region having the polypeptide sequence of SEQ ID NO: 167.Attorney Docket No. TP385350WO1 65. The antibody of any one of claims 59 to 64, wherein the antibody comprises a heavy chain having a polypeptide sequence of SEQ ID NO: 171 and a light chain having the polypeptide sequence of SEQ ID NO:

173.

66. The antibody of any one of claims 59 to 65, wherein the antibody comprises a heavy chain that is encoded by a polynucleotide sequence of SEQ ID NO: 170 and a light chain that is encoded by a polynucleotide sequence of SEQ ID NO:

172.

67. An antibody that is a Rabbit anti-Mouse IgG secondary antibody, wherein the antibody comprises a heavy chain (HC) and a light chain (LC) that pair to form a binding domain comprising a heavy chain complementarity determining region 1 (HCDR1) comprising the amino acid sequence of SEQ ID NO: 127, a HCDR2 comprising the amino acid sequence of SEQ ID NO: 128, a HCDR3 comprising the amino acid sequence of SEQ ID NO: 129, a light chain complementarity determining region 1 (LCDR1) comprising the amino acid sequence of SEQ ID NO: 132, a LCDR2 comprising the amino acid sequence of GAS, and a LCDR3 comprising the amino acid sequence of SEQ ID NO: 133, and wherein the heavy chain further comprises a first CH2 domain, a second CH2 domain, and a third CH2 domain, wherein the first CH2 domain and the second CH2 domain are connected by a first linker and an optional first hinge, and wherein the second CH2 domain and the third CH2 domain are connected by a second linker and an optional second hinge, and wherein the heavy chain further comprises a CH3 domain.

68. The antibody of claim 67, wherein the first CH2 domain, the second CH2 domain, and the third CH2 domain each comprises the amino acid sequence of SEQ ID NO: 19 or SEQ ID NO:

49.

69. The antibody of claim 67 or claim 68, wherein the first hinge and the second hinge each comprises the amino acid sequence of SEQ ID NO: 18 or SEQ ID NO:

48.

70. The antibody of any one of claims 67 to 69, wherein the first linker and the second linker each comprises the amino acid sequence of GS, GGS, SEQ ID NO: 14, SEQ ID NO: 38, SEQ ID NO: 43, or SEQ ID NO: 44.Attorney Docket No. TP385350WO1 71. The antibody of any one of claims 67 to 70, wherein the CH3 domain comprises the amino acid sequence of SEQ ID NO:

20.

72. The antibody of any one of claims 67 to 71, wherein the antibody comprises a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 126 and a light chain variable region having the polypeptide sequence of SEQ ID NO:

131.

73. The antibody of any one of claims 67 to 72, wherein the antibody comprises a heavy chain having a polypeptide sequence of SEQ ID NO: 135 and a light chain having the polypeptide sequence of SEQ ID NO:

137.

74. The antibody of any one of claims 67 to 73, wherein the antibody comprises a heavy chain that is encoded by a polynucleotide sequence of SEQ ID NO: 134 and a light chain that is encoded by a polynucleotide sequence of SEQ ID NO:

136.

75. An antibody that immunospecifically binds to pERK1 / 2, wherein the antibody comprises a heavy chain (HC) and a light chain (LC) that pair to form a binding domain comprising a heavy chain complementarity determining region 1 (HCDR1) comprising the amino acid sequence of SEQ ID NO: 85, a HCDR2 comprising the amino acid sequence of SEQ ID NO: 86, a HCDR3 comprising the amino acid sequence of SEQ ID NO: 87, a light chain complementarity determining region 1 (LCDR1) comprising the amino acid sequence of SEQ ID NO: 91, a LCDR2 comprising the amino acid sequence of KVS, and a LCDR3 comprising the amino acid sequence of SEQ ID NO: 92, and wherein the heavy chain further comprises a first CH2 domain, a second CH2 domain, and a third CH2 domain, wherein the first CH2 domain and the second CH2 domain are connected by a first linker and an optional first hinge, and wherein the second CH2 domain and the third CH2 domain are connected by a second linker and an optional second hinge, and wherein the heavy chain further comprises a CH3 domain.

76. The antibody of claim 75, wherein the first CH2 domain, the second CH2 domain, and the third CH2 domain each comprises the amino acid sequence of SEQ ID NO:Attorney Docket No. TP385350WO1 77. The antibody of claim 75 or claim 76, wherein the first hinge and the second hinge each comprises the amino acid sequence of SEQ ID NO:

102.

78. The antibody of any one of claims 75 to 77, wherein the first linker and the second linker each comprises the amino acid sequence of GS, GGS, SEQ ID NO: 14, SEQ ID NO: 38, SEQ ID NO: 43, or SEQ ID NO:

44.

79. The antibody of any one of claims 75 to 78, wherein the CH3 domain comprises the amino acid sequence of SEQ ID NO:

106.

80. The antibody of any one of claims 75 to 79, wherein the antibody comprises a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 83 and a light chain variable region having the polypeptide sequence of SEQ ID NO:

90.

81. The antibody of any one of claims 75 to 80, wherein the antibody comprises a heavy chain having a polypeptide sequence of SEQ ID NO: 94 and a light chain having the polypeptide sequence of SEQ ID NO:

96.

82. The antibody of any one of claims 75 to 81, wherein the antibody comprises a heavy chain that is encoded by a polynucleotide sequence of SEQ ID NO: 93 and a light chain that is encoded by a polynucleotide sequence of SEQ ID NO:

95.

83. An antibody that immunospecifically binds to PSD-95, wherein the antibody comprises a heavy chain (HC) and a light chain (LC) that pair to form a binding domain comprising a heavy chain complementarity determining region 1 (HCDR1) comprising the amino acid sequence of SEQ ID NO: 111, a HCDR2 comprising the amino acid sequence of SEQ ID NO: 112, a HCDR3 comprising the amino acid sequence of SEQ ID NO: 113, a light chain complementarity determining region 1 (LCDR1) comprising the amino acid sequence of SEQ ID NO: 116, a LCDR2 comprising the amino acid sequence of DTS, and a LCDR3 comprising the amino acid sequence of SEQ ID NO: 117, and wherein the heavy chain further comprises a first CH2 domain, a second CH2 domain, and a third CH2 domain, wherein the first CH2 domain and the second CH2 domain are connected by a first linker and anAttorney Docket No. TP385350WO1 optional first hinge, and wherein the second CH2 domain and the third CH2 domain are connected by a second linker and an optional second hinge, and wherein the heavy chain further comprises a CH3 domain.

84. The antibody of claim 83, wherein the first CH2 domain, the second CH2 domain, and the third CH2 domain each comprises the amino acid sequence of SEQ ID NO:

104.

85. The antibody of claim 83 or claim 84, wherein the first hinge and the second hinge each comprises the amino acid sequence of SEQ ID NO:

102.

86. The antibody of any one of claims 83 to 85 wherein the first linker and the second linker each comprises the amino acid sequence of GS, GGS, SEQ ID NO: 14, SEQ ID NO: 38, SEQ ID NO: 43, or SEQ ID NO:

44.

87. The antibody of any one of claims 83 to 86, wherein the CH3 domain comprises the amino acid sequence of SEQ ID NO:

106.

88. The antibody of any one of claims 83 to 87, wherein the antibody comprises a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 110 and a light chain variable region having the polypeptide sequence of SEQ ID NO:

115.

89. The antibody of any one of claims 83 to 88, wherein the antibody comprises a heavy chain having a polypeptide sequence of SEQ ID NO: 119 and a light chain having the polypeptide sequence of SEQ ID NO:

121.

90. The antibody of any one of claims 83 to 89, wherein the antibody comprises a heavy chain that is encoded by a polynucleotide sequence of SEQ ID NO: 118 and a light chain that is encoded by a polynucleotide sequence of SEQ ID NO:

120.

91. An antibody that immunospecifically binds to TIGIT, wherein the antibody comprises a heavy chain (HC) and a light chain (LC) that pair to form a binding domain comprising a heavy chain complementarity determining region 1 (HCDR1)Attorney Docket No. TP385350WO1 comprising the amino acid sequence of SEQ ID NO: 146, a HCDR2 comprising the amino acid sequence of SEQ ID NO: 147, a HCDR3 comprising the amino acid sequence of SEQ ID NO: 148, a light chain complementarity determining region 1 (LCDR1) comprising the amino acid sequence of SEQ ID NO: 151, a LCDR2 comprising the amino acid sequence of SAS, and a LCDR3 comprising the amino acid sequence of SEQ ID NO: 152, and wherein the heavy chain further comprises a first CH2 domain, a second CH2 domain, and a third CH2 domain, wherein the first CH2 domain and the second CH2 domain are connected by a first linker and an optional first hinge, and wherein the second CH2 domain and the third CH2 domain are connected by a second linker and an optional second hinge, and wherein the heavy chain further comprises a CH3 domain.

92. The antibody of claim 91, wherein the first CH2 domain, the second CH2 domain, and the third CH2 domain each comprises the amino acid sequence of SEQ ID NO:

19.

93. The antibody of claim 91 or claim 92, wherein the first hinge and the second hinge each comprises the amino acid sequence of SEQ ID NO:

18.

94. The antibody of any one of claims 91 to 93, wherein the first linker and the second linker each comprises the amino acid sequence of GS, GGS, SEQ ID NO: 14, SEQ ID NO: 38, SEQ ID NO: 43, or SEQ ID NO:

44.

95. The antibody of any one of claims 91 to 94, wherein the CH3 domain comprises the amino acid sequence of SEQ ID NO:

20.

96. The antibody of any one of claims 91 to 95, wherein the antibody comprises a heavy chain variable region having the polypeptide sequence of SEQ ID NO: 145 and a light chain variable region having the polypeptide sequence of SEQ ID NO:

150.

97. The antibody of any one of claims 91 to 96, wherein the antibody comprises a heavy chain having a polypeptide sequence of SEQ ID NO: 154 and a light chain having the polypeptide sequence of SEQ ID NO: 156.Attorney Docket No. TP385350WO1 98. The antibody of any one of claims 91 to 97, wherein the antibody comprises a heavy chain that is encoded by a polynucleotide sequence of SEQ ID NO: 153 and a light chain that is encoded by a polynucleotide sequence of SEQ ID NO:

155.

99. The antibody of any one of claims 13 to 32 or 35 to 98, wherein the antibody provides at least a 1.5-fold increase in signal enhancement compared to a corresponding wild type antibody.

100. The antibody of any one of claims 13 to 33 or 35 to 99, wherein the antibody provides at least a 1.5-fold reduction in background signal compared to a corresponding wild type antibody.

101. A method of detecting the presence of a target protein in a sample, said method comprising: applying the isolated polypeptide of any one of claims 1 to 12 or the antibody of any one of claims 13 to 33 or claims 35 to 100 to a sample containing a target protein, incubating the isolated polypeptide or the antibody in the sample under conditions permitting the isolated polypeptide or the antibody to bind to the target protein and to form a bound isolated polypeptide or a bound antibody, and determining the presence of the bound isolated polypeptide or the bound antibody, thereby detecting the presence of the target protein.

102. The method of claim 101, wherein the isolated polypeptide or the antibody has less background binding to cells that express Fc gamma receptors relative to a wild type isolated polypeptide or a wild type antibody.

103. The method of claim 101 or claim 102, wherein the target protein is OCT4, Parkin, IDO1, FLAG-tag, pERK1 / 2, PSD-95 or TIGIT.

104. The method of any one of claims 101 to 103, wherein the target protein is present in the sample in low abundance.Attorney Docket No. TP385350WO1 105. The method of any one of claims 101 to 104, wherein the presence of the bound isolated polypeptide or the bound antibody is determined by applying a secondary antibody to said sample, wherein the secondary antibody is labeled.

106. The method of any one of claims 101 to 105, wherein the antibody is labeled.

107. The method of claim 105 or claim 106, wherein the label is selected from a fluorophore, a fluorescent protein, a phosphorescent dye, a tandem dye, a polymer dye, a particle, an electron transfer agent, a hapten, an enzyme, a radioisotope, a chromophore, and an oligonucleotide-based fluorescent dye.

108. The method of claim 107, wherein the fluorophore is a pyrene, an anthracene, a naphthalene, an acridine, a stilbene, an indole or benzindole, an oxazole or benzoxazole, a thiazole or benzothiazole, an indacene, a 4-amino-7-nitrobenz-2-oxa- 1,3-diazole (NBD), a cyanine, a carbocyanine, a benzocyanine, a carbostyryl, a porphyrin, a salicylate, an anthranilate, an azulene, a perylene, a pyridine, a quinoline, a borapolyazaindacene, a xanthene, a rhodamine, a silicon rhodamine, a fluorescein, a rhodol, a rosamine, an oxazine or a benzoxazine, a benzophenoxazine, a carbazine, a phenalenone, a coumarin, a benzofuran, a benzophenalenone, a benzopyrilium, a styryl, a squarine, a resorufin, an anthraquinone, or a quinazolinone, or a conjugated polymer dye.

109. The method of claim 107 wherein the particle is a semiconductor nanocrystal.

110. The method of claim 107, wherein the hapten is a biotin or a biotin derivative.

111. The method of claim 107, wherein the enzyme is selected from a peroxidase, a phosphatase, a glycosidase, and a luciferase.

112. The method of claim 111, wherein the method further comprises adding an enzymatic substrate selected from a chromogenic, a fluorogenic, and a chemiluminescent substrate.

113. The method of any one of claims 106 to 112, wherein detecting the labeled antibody utilizes flow cytometry, immunohistochemistry, immunocytochemistry, or Western blotting.Attorney Docket No. TP385350WO1 114. The method of any one of claims 101 to 113, wherein the isolated polypeptide is an antibody with the isotype IgG.

115. A method of increasing sensitivity of an antibody, the method comprising: introducing into a heavy chain of the antibody a polypeptide comprising amino acid sequence of Formula I: Z-(X-Yn-Zr)p-X, wherein Z is a hinge; X is a CH2 domain; Y is a linker; n is an integer of 1 or greater; p is an integer of 2 or greater; and r is an integer of 0 or 1, wherein the sensitivity of the antibody is increased by at least 1.5-fold compared to a corresponding wild type antibody.

116. The method of claim 115, further comprising introducing LFLEPS mutations into the amino acid sequence of X, wherein the LFLEPS mutations comprise L234F, L235E and P331S substitutions.

117. The method of claim 115 or claim 116, wherein X comprises an amino acid sequence at least 90% identical to SEQ ID NO: 19, at least 90% identical to SEQ ID NO: 49, or at least 90% identical to SEQ ID NO:

104.

118. The method of any one of claims 115 to 117, wherein Z comprises an amino acid sequence at least 90% identical to SEQ ID NO: 18, at least 90% identical to SEQ ID NO: 48, or at least 90% identical to SEQ ID NO:

102.

119. The method of any one of claims 115 to 118, wherein Y comprises an amino acid sequence at least 90% identical to the amino acid sequence of GS, GGS, SEQ ID NO: 14, SEQ ID NO: 38, SEQ ID NO: 43, or SEQ ID NO:

44.

120. A method of reducing background signal of an antibody, the method comprising: introducing into a heavy chain of the antibody a polypeptide comprising amino acid sequence of Formula I: Z-(X-Yn-Zr)p-X, whereinAttorney Docket No. TP385350WO1 Z is a hinge; X is a CH2 domain; Y is a linker; n is an integer of 1 or greater; p is an integer of 2 or greater; and r is an integer of 0 or 1; wherein the background signal of the antibody is decreased by at least 1.5-fold compared to a corresponding wild type antibody.

121. The method of claim 120, further comprising introducing LFLEPS mutations into the amino acid sequence of X, wherein the LFLEPS mutations comprise L234F, L235E and P331S substitutions.

122. The method of claim 120 or claim 121, wherein X comprises an amino acid sequence at least 90% identical to SEQ ID NO: 19, at least 90% identical to SEQ ID NO: 49, or at least 90% identical to SEQ ID NO:

104.

123. The method of any one of claims 120 to claim 122, wherein Z comprises an amino acid sequence at least 90% identical to SEQ ID NO: 18, at least 90% identical to SEQ ID NO: 48, or at least 90% identical to SEQ ID NO:

102.

124. The method of any one of claims 120 to 123, wherein Y comprises an amino acid sequence at least 90% identical to the amino acid sequence of GS, GGS, SEQ ID NO: 14, SEQ ID NO: 38, SEQ ID NO: 43, or SEQ ID NO: 44.

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