ELP fusion of biologics and methods of making and using the same

Abstract

Disclosed herein are fusion proteins comprising a biologic and an elastin‑like polypeptide (ELP) attached to the biologic that is capable of being expressed and secreted from a eukaryotic cell. Also disclosed herein are methods for producing the biologic-based fusion protein.

Description

Docket No. 028193-0052-WO01ELP FUSION OF BIOLOGICS AND METHODS OF MAKING AND USING THE SAMECROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims priority to U. S. Provisional Patent Application No. 63 / 761,357, filed February 21, 2025 and U. S. Provisional Patent Application No. 63 / 883,805, filed September 18, 2025, each of which is incorporated herein by reference in its entirety.STATEMENT REGARDING FEDERALLY SPONSORED RESEARCH

[0002] This invention was made with government support under grant MURI 2834303 awarded by the U. S. Department of Defense, and grant R01 A034174 awarded by the National Institutes of Health. The government has certain rights in the invention.FIELD

[0003] This disclosure relates to a fusion protein comprising a biologic and an elastin-like polypeptide (ELP) attached to the biologic that is capable of being expressed and secreted from a eukaryotic cell. Also described herein are methods for producing the biologic-based fusion protein.INTRODUCTION

[0004] To date, fusions of an elastin-like polypeptide (ELP) fusions have solely been recombinantly expressed and purified from E. coli and only afterwards used as a reagent to purify antibodies. ELP-biologic fusions have not been developed before. The need to generate these constructs in bacterial systems which have to be purified before being used in antibody purification workflows increases the cost and complexity of antibody purification while simultaneously introducing potential contamination risks by carry-over endotoxin and other contaminants from the bacterial expression. This increases the risk of adverse effects especially during the administration of therapeutic antibodies.SUMMARY

[0005] In an aspect, the disclosure relates to a fusion protein comprising: a biologic; and an elastin-like polypeptide (ELP) attached to the biologic, wherein the ELP comprises an amino acid sequence of [VPGXG]m(SEQ ID NO: 1), wherein the ELP has a hydro path i city score of from about 0.5 to about 1.5 and m is an integer from 10 to 120, wherein the fusion protein isDocket No. 028193-0052-WO01expressed and secreted from a eukaryotic cell. In an embodiment, the ELP is attached to the C-terminus of the biologic. In another embodiment, one ELP is attached to the biologic. In another embodiment, more than one ELP is attached to the biologic. In another embodiment, the biologic is an antibody or a Fc-fusion protein. In another embodiment, the antibody is a monoclonal antibody, an immunoglobulin, or a bispecific antibody. In another embodiment, the ELP is attached to a terminus of a heavy chain of the antibody or a terminus of a light chain of the antibody. In another embodiment, the antibody comprises more than one ELP, each ELP being independently attached to a terminus of a heavy chain of the antibody, a terminus of a light chain of the antibody, or a combination thereof. In another embodiment, the antibody comprises two ELPs, a first ELP attached to a terminus of a first heavy chain and a second ELP attached to a terminus of a second heavy chain; the antibody comprises two ELPs, a first ELP attached to a terminus of a first light chain and a second ELP attached to a terminus of a second light chain; or the antibody comprises four ELPs, a first ELP attached to a terminus of a first heavy chain, a second ELP attached to a terminus of a second heavy chain, a third ELP attached to a terminus of a first light chain, and a fourth ELP attached to a terminus of a second light chain. In another embodiment, the Fc-fusion protein comprises one ELP attached to a terminus of a first heavy chain or a second heavy chain; or the Fc-fusion protein comprises two ELPs, a first ELP attached to a terminus of a first heavy chain and a second ELP attached to a terminus of a second heavy chain. In another embodiment, the Fc-fusion protein comprises a modified human IgG Fc domain. In another embodiment, the Fc-fusion protein comprises a p35 domain, a p40 domain, an interleukin-12 (IL-12) domain, or a combination thereof. In another embodiment, the ELP comprises an amino acid sequence of [VPGXG]m (SEQ ID NO: 1); wherein X is alanine (Ala), valine (Val), Serine (Ser), Isoleucine (Ile), or a mixture thereof; and wherein m is an integer from 20 to 120. In another embodiment, X is a mixture of Ala and Val in a ratio from 10:1 to 1:10 (Ala: Val). In another embodiment, X is a mixture of Ala and Val in a ratio from 4:1 to 1:4 (Ala: Val). In another embodiment, m is 40 and X is Val or a mixture of Ala and Val in a ratio from 1:1 to 1:4 (Ala: Val); or m is 80 and X is Ala or a mixture of Ala and Val in a ratio from 1:1 to 4:1 (Ala: Val). In another embodiment, X comprises Ser and lie arranged in blocks, each block comprising about 1 to about 40 repeats of a single residue, and wherein the overall ratio of Ser to He residues is from about 10:1 to about 1:10. In another embodiment, the ELP comprises an amino acid sequence of [VPGVG]4o(SEQ ID NO: 2); [VPGAG]4o (SEQ ID NO: 3); [VPG[A1: V1]G]40(SEQ ID NO: 4); [VPG[AI: V4]G]40(SEQ ID NO: 5); [VPG[A4: VI]G]40(SEQ ID NO: 6); [VPGAG]80(SEQ ID NO: 11); [VPGVG]80(SEQ ID NO: 12); [VPG[A1: V1]G]80(SEQ ID NO: 14); [VPG[A4: VI]G]80(SEQ ID NO: 15);[VPG[AI: V4]G]80(SEQ ID NO: 16); [VPGVG]120(SEQDocket No. 028193-0052-WO01ID NO: 21); [VPGSG]2o-[VPGIG]2o-[VPGSG]2o-[VPGIG]2o(SEQ ID NO: 17); [VPGSG]20-[VPGIG]2O-[VPGSG]2O-[VPGIG]2O-[VPGIG]2O(SEQ ID NO: 24); [VPGSG]8o (SEQ ID NO: 13); or [VPGIG]4O-[VPGSG]4O (SEQ ID NO: 115). In another embodiment, the fusion protein has a glycosylation pattern characteristic of the eukaryotic cell. In another embodiment, the ELP has a hydrophobicity effective to promote expression and secretion from the cell without causing precipitation or aggregation. In another embodiment, the ELP has a transition temperature (Tt) that is at least 2°C higher than a temperature that the at least one eukaryotic cell is cultured at. In another embodiment, the ELP has a transition temperature (Tt) of from about 34°C to about 65°C. In another embodiment, the ELP is capable of undergoing a reversible phase transition. In another embodiment, the fusion protein comprises a secondary protein domain attached to the C-terminus of the ELP, wherein the secondary protein domain comprises mCherry, maltose-binding protein (MBP), thioredoxin, N-utilization substance A (NusA), green fluorescent protein (GFP), glutathione S-transferase (GST), an engineered small ubiquitin-like modifier (SUMO) tag, a modified bacterial dehalogenase tag, human serum albumin (HSA), or a second Fc-fusion protein or domain thereof. In another embodiment, the fusion protein comprises a secretion domain attached to the N-terminus of the fusion protein. In another embodiment, the secretion domain comprises an azurocidin signal peptide (SEQ ID NO: 28), an interleukin-2 (IL-2) signal peptide (SEQ ID NO: 31), an albumin signal peptide (SEQ ID NO: 32), a CD33 signal peptide (SEQ ID NO: 33), a tissue plasminogen activator (tPA) signal peptide (SEQ ID NO: 34), a human trypsinogen-2 signal peptide (SEQ ID NO: 35), a human chymotrypsinogen signal peptide (SEQ ID NO: 36), a maltose-binding protein (MBP) signal peptide (SEQ ID NO: 37), a Fc Tag, a glutathione S-transferase (GST) Tag (SEQ ID NO: 40), a CD5 signal peptide (SEQ ID NO: 38), an immunoglobulin Kappa light chain signal peptide (SEQ ID NO: 39), a trypsinogen signal peptide (SEQ ID NO: 29), or a preprolactin signal peptide (SEQ ID NO: 30).

[0006] In a further aspect, the disclosure relates to a method of producing a biologic-based fusion protein, comprising: (a) engineering a eukaryotic cell to express a fusion protein comprising a biologic and an elastin-like polypeptide (ELP) attached to a C-terminus of the biologic, wherein the ELP comprises an amino acid sequence of [VPGXG]m(SEQ ID NO: 1), wherein the ELP has a hydropathicity score of from about 0.5 to about 1.5 and m is an integer from 10 to 120; (b) culturing the eukaryotic cell under conditions permitting secretion of the fusion protein into a culture supernatant; and (c) recovering the fusion protein from the culture supernatant. In an embodiment, the eukaryotic cell is a mammalian cell or an insect cell. In another embodiment, the mammalian cell is a human cell, a mouse cell, a hamster cell, a ratDocket No. 028193-0052-WO01cell, a goat cell, or a rabbit cell. In another embodiment, the fusion protein is secreted without intracellular aggregation. In another embodiment, the biologic is an antibody or an Fc-fusion protein. In another embodiment, the ELP comprises an amino acid sequence of [VPGXG]m(SEQ ID NO: 1); wherein X is alanine (Ala), valine (Val), Serine (Ser), Isoleucine (Ile), or a mixture thereof; and wherein m is an integer from 20 to 120. In another embodiment, X is a mixture of Ala and Val in a ratio from 10:1 to 1:10 (Ala: Val). In another embodiment, X is a mixture of Ala and Val in a ratio from 4:1 to 1:4 (Ala: Val). In another embodiment, m is 40 and X is Val or a mixture of Ala and Val in a ratio from 1:1 to 1:4 (Ala: Val); or m is 80 and X is Ala or a mixture of Ala and Val in a ratio from 1:1 to 4:1 (Ala: Val). In another embodiment, X comprises Ser and lie arranged in blocks, each block comprising about 1 to about 40 repeats of a single residue, and wherein the overall ratio of Ser to Ile residues is from about 10:1 to about 1:10. In another embodiment, the ELP has a hydrophobicity effective to promote expression and secretion from the cell without causing precipitation or aggregation. In another embodiment, the ELP has a transition temperature (Tt) that is at least 2°C higher than a temperature that the at least one eukaryotic cell is cultured at.

[0007] Another aspect of the disclosure provides a method of purifying a fusion protein, comprising: (a) providing a culture supernatant comprising the fusion protein described herein; (b) inducing a phase transition of the ELP to form an insoluble phase; (c) isolating the insoluble phase from the supernatant; and (d) resolubilizing the fusion protein. In an embodiment, the phase transition is induced by increasing temperature of the supernatant above a transition temperature (Tt) of the ELP. In another embodiment, the phase transition is induced by adding from about 0.05M to about 5M salt to the supernatant. In another embodiment, the salt is sodium chloride, ammonium sulfate, sodium citrate, sodium sulfate, magnesium sulfate, potassium sulfate, sodium phosphates, lithium chloride, sodium acetate, or sodium carbonate. In another embodiment, step (c) comprises centrifugation or tangential flow filtration. In another embodiment, step (d) comprises adding a low salt buffer to the fusion protein and removing insoluble contaminants using centrifugation or tangential flow filtration. In another embodiment, the low salt buffer is phosphate-buffered saline (PBS), Dulbecco’s Phosphate- Buffered Saline (DPBS), citrate buffer, 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (HEPES) buffer, tris(hydroxymethyl)aminomethane (Tris) buffer, acetate buffer, histidine buffer, or succinate buffer. In another embodiment, the method is performed without chromatography.

[0008] Another aspect of the disclosure provides a method of treating a disease or disorder, comprising administering to a subject a therapeutically effective amount of the fusion proteinDocket No. 028193-0052-WO01described herein. In an embodiment, the fusion protein exhibits extended serum half-life relative to the biologic without the ELP. In another embodiment, the fusion protein is administered subcutaneously, intramuscularly, or intratumorally; and forms a depot at the administration site. In another embodiment, the disease or disorder is cancer, an autoimmune disease, or a metabolic disorder.

[0009] Another aspect of the disclosure provides a kit, comprising: (a) a fusion protein described herein; and (b) instructions for administering or purifying the fusion protein. In an embodiment, the kit further comprises a buffer suitable for inducing or reversing a phase transition of the ELP, or suitable for in vivo administration.

[0010] The disclosure provides for other aspects and embodiments that will be apparent in light of the following detailed description and accompanying figures.BRIEF DESCRIPTION OF THE DRAWINGS

[0011] FIGS. 1A-B show ELPs fused to a human therapeutic antibody (Trastuzumab). FIG.1A is a diagram showing a depiction of the different investigated architectures, where ELPs are fused to the C-terminus of the heavy and / or light chains. FIG. 1B is an image of SPS-PAGE Gels run under non-reducing (NR) and reducing (R) conditions. In gel 1: Lane 1 is the protein ladder. A - native Trastuzumab (lanes 2 and 3), B - Trastuzumab-ELP where a V(40) ELP (SEQ ID NO: 42) is fused to the antibody heavy chain (HC) (lanes 4 and 5), C - Trastuzumab-ELP where a V(40) ELP (SEQ ID NO: 43) is fused to the antibody light chain (LC) (lanes 6 and 7), D - Trastuzumab-ELP where a V(40) ELP (SEQ ID NO: 44) is fused to the antibody HC and LC (lanes 8 and 9), E - Trastuzumab-ELP where a V4A1(40) ELP (SEQ ID NO: 45) is fused to the antibody (HC) (lanes 10 and 11), F - Trastuzumab-ELP where a V4A1(40) ELP (SEQ ID NO: 46) is fused to the antibody LC (lanes 12 and 13), G - Trastuzumab-ELP where a V4A1(40) ELP (SEQ ID NO: 47) is fused to the antibody HC and LC (lanes 14 and 15). In gel 2: Lane 11 is the protein ladder. H -Trastuzumab-ELP where a V1A1(40) ELP (SEQ ID NO: 48) is fused to the antibody HC (lanes 1 and 2), I - Trastuzumab-ELP where a V1A1(40) ELP (SEQ ID NO: 49) is fused to the antibody LC (lanes 3 and 4), J - Trastuzumab-ELP where a V1A1(40) ELP (SEQ ID NO: 50) is fused to the antibody HC and LC (lanes 5 and 6), K- Trastuzumab-ELP where a V1A4(40) ELP (SEQ ID NO: 51) is fused to the antibody HC (lanes 7 and 8), L -Trastuzumab-ELP where a V1A4(40) ELP (SEQ ID NO: 52) is fused to the antibody LC (lanes 9 and 10). In gel 3: Lane 9 is the protein ladder. M - Trastuzumab-ELP where a V1A4(40) ELPDocket No. 028193-0052-WO01(SEQ ID NO: 51) is fused to the antibody LC (lanes 1 and 2), N -Trastuzumab-ELP where a A(40) ELP (SEQ ID NO: 54) is fused to the antibody HC (lanes 3 and 4), O - Trastuzumab-ELP where a A(40) ELP (SEQ ID NO: 55) is fused to the antibody LC (lanes 5 and 6), P -Trastuzumab-ELP where a A(40) ELP (SEQ ID NO: 56) is fused to the antibody HC and LC (lanes 7 and 8).

[0012] FIG. 2 is a graph showing yields in nanomolar of different T rastuzumab-ELP fusions. Constructs were expressed in Expi293 cells for 7 days at 37°C. Trastuzumab and Trastuzumab-ELP fusions were purified with a Protein A chromatography column.

[0013] FIG. 3 is a graph showing half-maximal effective concentration (EC50) assays that were performed by chemiluminescent ELISA. A 1 g / mL solution of HER2 in bicarbonate buffer was incubate in MaxiSorp plates overnight. Next, plates were washed 3 times with IxTBST buffer and blocked with a 2% BSA in IxTBST for 1 hour. After 3 washes (IxTBST), microwells were incubated with 100 L of trastuzumab or trastuzumab-ELP solutions with various concentrations ranging from 0.03 nM to 100 nM in 1xPBS in triplicate. Following 3 washes (IxTBST), wells were incubated for 1-hour with an anti-human IgG antibody conjugated to HRP and washed with IxTBST 3 more times. Finally, wells were incubated with a supersignal chemiluminescent substrate and imaged at 425nm.

[0014] FIGS. 4A-B show guanidine-HCI denaturation assays in a subset of Trastuzumab-ELPs versus the unmodified T rastuzumab. T rastuzumab or trastuzumab-ELP at a final concentration of 0.5 M was denatured overnight at room temperature in guanidine-HCI at a pH = 7.4 and concentration varying from 0 to 5.5M in 0.5M increments. Assays were done in triplicate in clear bottom plates. Intrinsic tryptophan fluorescence was detected by exciting the samples at 280nm and ratio of 350nm to 330nm was acquired. Data was fitted to a two-state model. FIG. 4A is a graph showing fluorescence signal. FIG. 4B is a table showing the data.

[0015] FIGS. 5A-B show Trastuzumab-ELP HC V(40) (SEQ ID NO: 42) production in ExpiCHO cells and Transition temperature (Tt) characterization. FIG. 5A is an image of a SPS-PAGE gel under reducing and non-reducing conditions (lanes 2 and 3) of Protein A and SEC purified Trastuzumab-ELP HC V(40) (SEQ ID NO: 42). FIG. 5B is a graph showing transition temperature characterization for Trastuzumab-ELP HC V(40) (SEQ ID NO: 42) where absorbance at 350nm was collected at different temperatures generating heating (solid line) and cooling (dashed line) curves in 0.6°C intervals at four different micromolar concentrations.Docket No. 028193-0052-WO01

[0016] FIGS. 6A-B show Trastuzumab-ELP HC V4A1(80) (SEQ ID NO: 62) production in ExpiCHO cells and Transition temperature (Tt) characterization. FIG. 6A is an image showing a SPS-PAGE gel under reducing and non-reducing conditions (lanes 2 and 3) of Protein A and SEC purified Trastuzumab-ELP HC V4A1(80) (SEQ ID NO: 62). FIG. 6B is a graph showing transition temperature characterization for Trastuzumab-ELP HC V4A1(80) (SEQ ID NO: 62) where absorbance at 350nm was collected at different temperatures generating heating (solid line) and cooling (dashed line) curves in 0.6°C intervals at four different micromolar concentrations.

[0017] FIGS. 7A-B show ELPs fused to a murine antibody (“Mouse Antibody”) light (LC) and heavy chains (HC). FIG. 7A is a diagram showing: i. depiction of PCDNA5 plasmids containing HC and LC elements, where LC is fused to SISI80 ELP (SEQ ID NO: 102); ii. depiction of the Mouse Antibody mAb with ELP fused LC-terminal region; and iii. SDS-Page gel with ladder on lane (1), non-reduced product of protein A purification where light gray arrow points to mAb-ELP on lane (2) and reduced product of protein A purification where medium gray arrow points to HC and black arrow points to LC-SISI80 (SEQ ID NO: 102). FIG. 7B is a diagram showing: i. depiction of PCDNA5 plasmids containing HC and LC elements, where HC is fused to SISI80 ELP (SEQ ID NO: 100); ii. depiction of the Mouse Antibody mAb with ELP fused HC-terminal region; and iii. SDS-Page gel with ladder on lane (1), non-reduced product of protein A purification where light gray arrow points to mAb-ELP on lane (2) and reduced product of protein A purification where medium gray arrow points to HC-SISI80 (SEQ ID NO: 100) and black arrow points to LC. Abbreviations: HC = heavy chain; LC = light chain; mlgG2a = mouse immunoglobin a; VL = variable light chain; CL = constant light chain; VH = variable heavy chain; CH1 constant heavy chain 1; CH2 constant heavy chain 2; CH3 constant heavy chain 3; Fc = fragment crystallizable region.

[0018] FIGS. 8A-C show ELPs fused to a murine antibody (Mouse Antibody) LC and HC simultaneously and to the mAb HC and to a fluorescent protein (mCherry). FIG. 8A is a diagram showing: i. depiction of PCDNA5 plasmids containing HC and LC elements, where HC and LC are fused to SISI80 ELP (SEQ ID NOs: 100 & 102) and ii. depiction of the Mouse Antibody mAb with ELP fused to HC and LC terminal regions. FIG. 8B is a diagram showing: i. depiction of PCDNA5 plasmids containing HC and LC elements, where HC is fused to S80 ELP (SEQ ID NO: 106) and to mCherry fluorescent protein and ii. depiction of the Mouse Antibody mAb with ELP-mCherry fused to the C-terminus of the HC-ELP region. FIG. 8C is an image of a SDS-PAGE gel with MW ladder in lane (1), non-reduced and reduced products of protein ADocket No. 028193-0052-WO01and SEC purification of Mouse Antibody (HC-SISI80 + LC) in lanes (2-3), non-reduced and reduced products of protein A and SEC purification of Mouse Antibody (HC-SISI80 + LC-SISI80) in lanes (4-5) and non-reduced and reduced products of protein A and SEC purification of Mouse Antibody (HC-SISI80-mCherry + LC) in lanes (6-7). Abbreviations: HC = heavy chain; LC = light chain; mlgG2a - mouse immunoglobin a; VL = variable light chain; CL = constant light chain; VH = variable heavy chain; CH1 constant heavy chain 1; CH2 constant heavy chain 2; CH3 constant heavy chain 3; Fc = fragment crystallizable region; kDa = kilodalton.

[0019] FIGS. 9A-D show ELPs with different sizes and architectures fused to Dulaglutide.FIG. 9A is a diagram showing PCDNA5 plasmids containing Dulaglutide and Dulaglutide fused to various ELPs with sizes ranging from 40 to 120 repeats and different architectures. FIG. 9B is an image of a SDS-PAGE gel with ladder on lane (1), non-reduced and reduced products of protein A purification of Dulaglutide on lanes (2-3). FIG. 9C is an image of a SDS-PAGE gel with ladder in lane (1), non-reduced and reduced products of protein A purification of Dulaglutide-SI40 (SEQ ID NO: 79) on lanes (2-3), Dulaglutide-ISIS80 (SEQ ID NO: 80) in lanes (4-5), Dulaglutide-SSII80 (SEQ ID NO: 81) in lanes (6-7) and Dulaglutide-IISS80 (SEQ ID NO: 82) in lanes (8-9). FIG. 9D is an image of a SDS-PAGE gel with ladder in lane (1), non-reduced and reduced products of protein A purification of Dulaglutide-SISI80 in lanes (2-3), Dulaglutide-SISIS1120 (SEQ ID NO: 83) in lanes (4-5). Abbreviations: mhlgG4 = mutated human immunoglobin 4; CH2 constant heavy chain 2; CH3 constant heavy chain 3; Fc = fragment crystallizable region. GLP-1a = glucagon like peptide receptor agonist; kDa = kilodalton.

[0020] FIG. 10 is an image of a SDS-PAGE gel showing ELPs with of different sizes with valine as guest residue fused to Dulaglutide. SDS-PAGE gel unmodified Dulaglutide under nonreducing and reducing conditions in lanes 1 and 2, protein ladder on lane 3, Dulaglutide-ELP fusions run under non-reduced and reduced conditions in lanes 4 through 9.

[0021] FIGS. 11A-B show expression of Trastuzumab and ELP fused to Trastuzumab’s light chain. FIG. 11A is a diagram showing: i. depiction of PCDNA5 plasmids containing HC and LC sequences that code for Trastuzumab; ii. depiction of Trastuzumab HC and LC; and iii. SDS-PAGE gel with MW ladder on lane (1), non-reduced products of protein A purification where the unbound fraction is shown on lane (2), column wash on lane (3) and purified Trastuzumab on lane which is pointed by gray arrow (4). FIG. 11B is a diagram showing: i. depiction of PCDNA5 plasmids containing sequences that code for Trastuzumab HC and LC fused to IISS80 ELP (SEQ ID NO: 72); ii. depiction of Trastuzumab HC and LC-IISS80 (SEQ ID NO: 72); and iii.Docket No. 028193-0052-WO01SDS-PAGE gel with MW ladder on lane (4), non-reduced products of protein A purification where the unbound fraction is shown on lane (1), column wash on lane (2) and purified Trastuzumab-ELP on lane (3) which is pointed by gray arrow. Abbreviations: HC = heavy chain; LC = light chain; hlgG 1 = human immunoglobin 1; VL = variable light chain; CL = constant light chain; VH = variable heavy chain; CH1 constant heavy chain 1; CH2 constant heavy chain 2; CH3 constant heavy chain 3; Fc = fragment crystallizable region; kDa = kilodalton.

[0022] FIGS. 12A-C show expression of ELP fused to Trastuzumab’s HC. FIG. 12A is a diagram showing a depiction of PCDNA5 plasmids containing sequences that encode for Trastuzumab HC fused to IISS80 (SEQ ID NO: 72) and LC. FIG. 12B is a diagram showing a depiction of Trastuzumab HC-IISS80 (SEQ ID NO: 70). FIG. 12C is an image of a SDS-PAGE gel with ladder in lane (1), non-reduced and reduced products of protein A purification, where lanes (2-3) are Trastuzumab run as a control and lanes (4-5) are Trastuzumab-ELP, where the ELP is IISS80 (SEQ ID NO: 70) is fused to the HC of Trastuzumab. Abbreviations: HC = heavy chain; LC = light chain; hlgG1 = human immunoglobin 1; VL = variable light chain; CL = constant light chain; VH = variable heavy chain; CH1 constant heavy chain 1; CH2 constant heavy chain 2; CH3 constant heavy chain 3; Fc = fragment crystallizable region; kDa = kilodalton.

[0023] FIGS. 13A-E show phase transition behavior characterization of Dulaglutide and Dulaglutide-ELP fusions, where the absorbance at 350 nm is measured as a function of the solution temperature at different protein concentrations. FIG. 13A is a graph showing Dulaglutide. FIG. 13B is a graph showing Dulaglutide-SI40 (SEQ ID NO: 79). FIG. 13C is a graph showing Dulaglutide-ISIS80 (SEQ ID NO: 80). FIG. 13D is a graph showing Dulaglutide-SSII80 (SEQ ID NO: 81). FIG. 13E is a graph showing Dulaglutide-IISS80 (SEQ ID NO: 82). The legend from FIG. 13A can be applied to FIGS. 13B-E.

[0024] FIGS. 14A-D show phase transition behavior characterization of Dulaglutide and Dulaglutide-ELP fusions, where the absorbance at 350 nm is measured as a function of the solution temperature at different protein concentrations. FIG. 14A is a graph showing the various heating (solid line) and cooling (dashed lines) curves of Dulaglutide-V40 (SEQ ID NO: 86). FIG. 14B is a graph showing the various heating (solid line) and cooling (dashed lines) curves of Dulaglutide-V80 (SEQ ID NO: 87). FIG. 14C is a graph showing the various heating (solid line) and cooling (dashed lines) curves of Dulaglutide-V120 (SEQ ID NO: 88). FIG. 14D isDocket No. 028193-0052-WO01a graph showing the transition temperature dependency with concentration for the various valine based Dulaglutide-ELP fusions.

[0025] FIGS. 15A-D show inverse transition cycling (ITC) purification of mAb and FcfP fused to ELPs. FIG. 15A is a schematic showing ITC of ELP-fused mAbs and FcfPs, where cells, cell debris and insoluble proteins are removed from cell culture supernatant by centrifugation of depth filtration. Next, the supernatant is recovered, and the LCST phase transition is triggered by addition of salt. Following one round of centrifugation, the pellet contains the majority of the ELP fusion protein and the supernatant containing soluble contaminants is discarded. Cold, low salt buffer is added to reverse the phase transition and dissolve the pellet, recovering the purified mAb-ELP or FcfP-ELP. One extra round of cold centrifugation removes any other insoluble proteins and aggregates. Repeated cycles of ITC yield proteins with high degree of purity. ITC can be adapted for large volume purification at industrial scale by replacing centrifugation with tangential flow filtration. FIG. 15B is an image of a SDS-PAGE gel of ITC purification of Mouse Antibody-ELP, where SISI80 ELP (SEQ ID NO: 102) is fused to the C-terminus of the LC. Lane (1) is the ladder, lanes (2-3) are the dialyzed and filtered media prior to purification under non-reducing and reducing conditions respectively. Lanes (4-5) are the supernatant of the hot spin from the first cycle of ITC under non-reducing and reducing conditions, respectively. Lanes (6-7) are the cold spin after the first cycle of ITC, under non-reducing and reducing conditions, respectively, where the gray and black arrows point to the mAb and the HC and LC. Lanes (8-9) and (10-11) are the hot and cold spins after the second cycle of ITC under non-reducing and reducing conditions, respectively, where the gray and black arrows point to the purified mAb and the HC and LC. FIG. 15C is an image of a SDS-PAGE gel of ITC purification of Dulaglutide SISI80 (SEQ ID NO: 84), where lane (6) is the ladder and lane (1) is the dialyzed and filtered media under non-reducing conditions. Lanes 2-5 are the hot and cold spin supernatants after the first and second cycles of ITC under nonreducing conditions. Gray arrows highlight the Dulaglutide-ELP throughout the ITC purification.FIG. 15D is an image of a SDS-PAGE gel comparing Protein A and ITC purified Dulaglutide-ELP. Lane (5) is the ladder and lanes (1-2) and (3-4) are respectively the protein A and ITC purified Dulaglutide-ELP under non-reducing and reducing conditions, respectively.

[0026] FIGS. 16A-C show affinity measurements of Mouse Antibody and Mouse Antibody-ELPs binding to sGP by SPR. FIG. 16A is a graph of a sensorgram of the wild-type Mouse Antibody mAb. FIG. 16B is a graph of a sensorgram of Mouse Antibody with the SISI80 ELP (SEQ ID NO: 102) fused to the LC C-terminus. FIG. 16C is a graph of a sensorgram of MouseDocket No. 028193-0052-WO01Antibody with the SISI80 ELP (SEQ ID NO: 100) fused to the HC C-terminus. Data were fit to a 1:1 Langmuir model. Equilibrium dissociation constants (KD) is reported in nM.

[0027] FIGS. 17A-D show FcRn binding studies with WT hlgG4 and Dulaglutide. FIG. 17A is a graph showing a sensorgram collected with different concentrations of WT hlgG4 injected on a FcRn coated CM5 chip. Data were fit with a heterogenous ligand model (HLM). FIG. 17B is a graph showing sensorgram data for WT hlgG4Fc fit with a steady state ligand model. FIG.17C is a graph showing sensorgram collected with different concentrations of Dulaglutide injected on a FcRn coated CM5 chip. Data were fit with a heterogenous ligand model (HLM). FIG. 17D is a graph showing sensorgram data for Dulaglutide fit with a steady state ligand model. The equilibrium dissociation constant (KD) is reported in nanomolar (nM).Abbreviations: WT hlgG4Fc = wild type human immunoglobin 4 Fc; mhlgG4Fc = mutated human immunoglobin 4 Fc; Fc = fragment crystallizable region; KD = binding affinity.

[0028] FIGS. 18A-F show FcRn binding studies with Dulaglutide SI40 (SEQ ID NO: 79), Dulaglutide IISS80 (SEQ ID NO: 82), and Dulaglutide ISIS80 (SEQ ID NO: 80). FIG. 18A is a graph showing a sensorgram collected with different concentrations of Dulaglutide SI40 (SEQ ID NO: 79) injected on a FcRn coated CM5 chip. Data were fit with a heterogenous ligand model (HLM). FIG. 18B is a graph showing sensorgram data for Dulaglutide SI40 (SEQ ID NO: 79) fit with a steady state ligand model. FIG. 18C is a graph showing a sensorgram collected with different concentrations of Dulaglutide IISS80 (SEQ ID NO: 82) injected on a FcRn coated CM5 chip. Data were fit with a heterogenous ligand model (HLM). FIG. 18D is a graph showing sensorgram data for Dulaglutide IISS80 (SEQ ID NO: 82) fit with a steady state ligand model.FIG. 18E is a graph showing a sensorgram collected with different concentrations of Dulaglutide ISIS80 (SEQ ID NO: 80) injected on a FcRn coated CM5 chip. Data were fit with a heterogenous ligand model (HLM). FIG. 18F is a graph showing sensorgram data for Dulaglutide ISIS80 (SEQ ID NO: 80) fit with a steady state ligand model. The equilibrium dissociation constant (KD) is reported in nanomolar (nM). Abbreviations: WT hlgG4Fc = wild type human immunoglobin 4 Fc; mhlgG4Fc = mutated human immunoglobin 4 Fc; Fc = fragment crystallizable region; KD = binding affinity.

[0029] FIGS. 19A-D show FcRn binding studies with Dulaglutide SISI80 (SEQ ID NO: 84) and Dulaglutide SISIS1120 (SEQ ID NO: 83). FIG. 19A is a graph showing a sensorgram collected with different concentrations of Dulaglutide SISI80 (SEQ ID NO: 84) and C) Dulaglutide SISIS1120 injected on a FcRn coated CM5 chip. Data was fitted with aDocket No. 028193-0052-WO01heterogenous ligand model (HLM). The equilibrium dissociation constant (KD) is reported in nanomolar (nM). FIG. 19B is a graph showing sensorgram data for Dulaglutide SISI80 (SEQ ID NO: 84) fit with a steady state ligand model. The equilibrium dissociation constant (KD) is reported in nanomolar (nM). FIG. 19C is a graph showing a sensorgram collected with different concentrations of Dulaglutide SISIS1120 (SEQ ID NO: 83) injected on a FcRn coated CM5 chip. Data was fitted with a heterogenous ligand model (HLM). The equilibrium dissociation constant (KD) is reported in nanomolar (nM). FIG. 19D is a graph showing sensorgram data for Dulaglutide SISIS1120 (SEQ ID NO: 83) fit with a steady state ligand model. The equilibrium dissociation constant (KD) is reported in nanomolar (nM). Abbreviations: WT hlgG4Fc = wild type human immunoglobin 4 Fc; mhlgG4Fc = mutated human immunoglobin 4 Fc; Fc = fragment crystallizable region.

[0030] FIG. 20 is a graph showing in vitro activity of Dulaglutide ELP fusions.Chemiluminesce based ELISA with GLP-1 receptor on the surface of microwells was employed to determine ELP fusion activity. Various concentrations of Dulaglutide and Dulaglutide fusions were incubated in triplicate and developed with an HRP conjugated anti-human antibody and Supersignal luminescent substrate. Luminescence values were normalized to zero drug controls and data was fitted to a three parameter dose-response curve to determine EC50.

[0031] FIGS. 21A-C show characterization of example fusion proteins including p35 and p40 domains. FIG. 21A is an SDS PAGE gel of a final purified fusion protein of p35-Fc-V120 / p40-Fc where the ELP is attached to the p35 domain. FIG. 21 B is an SDS PAGE gel of a final purified fusion of p35-Fc-V120 / p40-Fc where the ELP is attached to the p40 domain. FIG.21 C is an SDS PAGE gel of a final purified fusion of p35-Fc-V120 / p40-Fc where an ELP is attached to the p35 domain and an ELP is attached to the p40 domain.DETAILED DESCRIPTION

[0032] Described herein are fusion proteins that include a biologic (e.g. Trastuzumab and Dulaglutide) fused to elastin-like polypeptides (ELPs) that can be generated and purified) in mammalian cells. Uniquely, the present application found a balance between hydrophobicity and hydrophilicity of the ELP that can allow the fusion protein to be expressed and secreted from mammalian cells. Mammalian cells are engineered to express and secrete monoclonal antibodies (mAb) and Fc fused proteins (FcfP) attached to ELPs into the cell-culture supernatant. When mAbs and FcfPs are expressed as an ELP fusion as described herein, theyDocket No. 028193-0052-WO01can be subsequently purified by standard methods such as Protein A chromatography, but also through reversible phase separation — a behavior driven by the ELP — of the mAb-ELP and FcfP-ELP by the addition of salt followed by centrifugation and / or filtration. By controlling the ELP’s architecture and composition, the mAb-ELP and FcfP-ELP can also be engineered to: control overall pharmacokinetics (PK) of mAbs and FcfPs — specifically, by reducing mAb and FcfP clearance and thereby extending their circulation times; form subcutaneous (s.c.) depots for controlled release of therapeutics; and facilitate s.c. delivery of biologic drugs by improving solubility of therapeutics.1. Definitions

[0033] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art. The meaning and scope of the terms should be clear. In case of conflict, the present document, including definitions, take precedent over any dictionary or extrinsic definition. Further, unless otherwise required by context, singular terms shall include pluralities and plural terms shall include the singular. Preferred methods and materials are described below, although methods and materials similar or equivalent to those described herein can be used in practice or testing of the present disclosure. All publications, patent applications, patents and other references mentioned herein are incorporated by reference in their entirety. The materials, methods, and examples disclosed herein are illustrative only and not intended to be limiting.

[0034] The terms “comprise(s),” “include(s),” “having,” “has,” “can,” “contain(s),” and variants thereof, as used herein, are intended to be open-ended transitional phrases, terms, or words that do not preclude the possibility of additional acts or structures. The singular forms “a,” “and,” and “the” include plural references unless the context clearly dictates otherwise. The present disclosure also contemplates other embodiments “comprising,” “consisting of,” and “consisting essentially of,” the embodiments or elements presented herein, whether explicitly set forth or not.

[0035] For the recitation of numeric ranges herein, each intervening number there between with the same degree of precision is explicitly contemplated. For example, for the range of 6-9, the numbers 7 and 8 are contemplated in addition to 6 and 9, and for the range 6.0-7.0, the number 6.0, 6.1, 6.2, 6.3, 6.4, 6.5, 6.6, 6.7, 6.8, 6.9, and 7.0 are explicitly contemplated.Docket No. 028193-0052-WO01

[0036] The term “about” or “approximately” as used herein as applied to one or more values of interest, refers to a value that is similar to a stated reference value, or within an acceptable error range for the particular value as determined by one of ordinary skill in the art, which will depend in part on how the value is measured or determined, such as the limitations of the measurement system. In certain aspects, the term “about” refers to a range of values that fall within 20%, 19%, 18%, 17%, 16%, 15%, 14%, 13%, 12%, 11%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, or less in either direction (greater than or less than) of the stated reference value unless otherwise stated or otherwise evident from the context (except where such number would exceed 100% of a possible value). Alternatively, “about” can mean within 3 or more than 3 standard deviations, per the practice in the art. Alternatively, such as with respect to biological systems or processes, the term “about” can mean within an order of magnitude, preferably within 5-fold, and more preferably within 2-fold, of a value.

[0037] “Affinity” refers to the binding strength of a polypeptide or protein to its target (i.e., binding partner).

[0038] “Agonist” refers to an entity that binds to a receptor and activates the receptor to produce a biological response. An “antagonist” blocks or inhibits the action or signaling of the agonist. An “inverse agonist” causes an action opposite to that of the agonist. The activities of agonists, antagonists, and inverse agonists may be determined in vitro, in situ, in vivo, or a combination thereof.

[0039] “Amino acid” as used herein refers to naturally occurring and non-natural synthetic amino acids, as well as amino acid analogs and amino acid mimetics that function in a manner similar to the naturally occurring amino acids. Naturally occurring amino acids are those encoded by the genetic code. Amino acids can be referred to herein by either their commonly known three-letter symbols or by the one-letter symbols recommended by the IUPAC-IUB Biochemical Nomenclature Commission. Amino acids include the side chain and polypeptide backbone portions.

[0040] “Coding sequence” or “encoding nucleic acid” as used herein means the nucleic acids (RNA or DNA molecule) 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 directingDocket No. 028193-0052-WO01expression in the cells of an individual or mammal to which the nucleic acid is administered. The coding sequence may be codon optimized.

[0041] “Complement” or “complementary” as used herein means a nucleic acid can mean Watson-Crick (e.g., A-T / U and C-G) or Hoogsteen base pairing between nucleotides or nucleotide analogs of nucleic acid molecules. “Complementarity” refers to a property shared between two nucleic acid sequences, such that when they are aligned antiparallel to each other, the nucleotide bases at each position will be complementary.

[0042] The terms “control,” “reference level,” and “reference” are used herein interchangeably. The reference level may be a predetermined value or range, which is employed as a benchmark against which to assess the measured result. “Control group” as used herein refers to a group of control subjects. The predetermined level may be a cutoff value from a control group. The predetermined level may be an average from a control group. The healthy or normal levels or ranges for a target or for a protein activity may be defined in accordance with standard practice. A control may be a subject or cell without a fusion as detailed herein. A control may be a subject, or a sample therefrom, whose disease state is known. The subject, or sample therefrom, may be healthy, diseased, diseased prior to treatment, diseased during treatment, or diseased after treatment, or a combination thereof.

[0043] “Functional” and “full-functional” as used herein describes protein that has biological activity. A “functional gene” refers to a gene transcribed to mRNA, which is translated to a functional protein.

[0044] “Fusion protein” as used herein refers to a chimeric protein created through the joining of two or more genes that originally coded for separate proteins. The translation of the fusion gene results in a single polypeptide with functional properties derived from each of the original proteins. A fusion protein as described herein may include a biologic and an ELP. The fusion protein may optionally include other domains. In one aspect, the ELP and fusion protein thereof has controlled reversible phase separation.

[0045] “Genetic construct” as used herein refers to the DNA or RNA molecules that comprise a polynucleotide that encodes a protein. The coding sequence includes initiation and termination signals operably linked to regulatory elements including a promoter and polyadenylation signal capable of directing expression in the cells of the individual to whom the nucleic acid molecule is administered. As used herein, the term “expressible form” refers toDocket No. 028193-0052-WO01gene constructs that contain the necessary regulatory elements operable linked to a coding sequence that encodes a protein such that when present in a cell, the coding sequence will be expressed.

[0046] The term “heterologous” as used herein refers to nucleic acid comprising two or more subsequences that are not found in the same relationship to each other in nature. For instance, a nucleic acid that is recombinantly produced typically has two or more sequences from unrelated genes synthetically arranged to make a new functional nucleic acid, for example, a promoter from one source and a coding region from another source. The two nucleic acids are thus heterologous to each other in this context. When added to a cell, the recombinant nucleic acids would also be heterologous to the endogenous genes of the cell. Thus, in a chromosome, a heterologous nucleic acid would include a non-native (non-naturally occurring) nucleic acid that has integrated into the chromosome, or a non-native (non-naturally occurring) extrachromosomal nucleic acid. Similarly, a heterologous protein indicates that the protein comprises two or more subsequences that are not found in the same relationship to each other in nature (for example, a “fusion protein,” where the two subsequences are encoded by a single nucleic acid sequence).

[0047] The term “host cell” is a cell that is susceptible to transformation, transfection, transduction, conjugation, and the like with a nucleic acid construct or expression vector. Host cells can be derived from plants, bacteria, yeast, fungi, insects, animals, etc. In some embodiments, the host cell includes a mammalian cell or an insect cell.

[0048] “Identical” or “identity” as used herein in the context of two or more polynucleotide or polypeptide sequences means that the sequences have a specified percentage of residues that are the same over a specified region. The percentage may 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) mayDocket No. 028193-0052-WO01be considered equivalent. Identity may be performed manually or by using a computer sequence algorithm such as BLAST or BLAST 2.0.

[0049] “Nucleic acid” or “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 polynucleotide also encompasses the complementary strand of a depicted single strand. Many variants of a polynucleotide may be used for the same purpose as a given polynucleotide. Thus, a polynucleotide also encompasses substantially identical polynucleotides and complements thereof. A single strand provides a probe that may hybridize to a target sequence under stringent hybridization conditions. Thus, a polynucleotide also encompasses a probe that hybridizes under stringent hybridization conditions. Polynucleotides may be single stranded or double stranded or may contain portions of both double stranded and single stranded sequence. The polynucleotide can be nucleic acid, natural or synthetic, DNA, genomic DNA, cDNA, RNA, or a hybrid, where the polynucleotide can contain combinations of deoxyribo- and ribo-nucleotides, and combinations of bases including, for example, uracil, adenine, thymine, cytosine, guanine, inosine, xanthine hypoxanthine, isocytosine, and isoguanine. Polynucleotides can be obtained by chemical synthesis methods or by recombinant methods.

[0050] “Open reading frame” refers to a stretch of codons that begins with a start codon and ends at a stop codon. In eukaryotic genes with multiple exons, introns are removed, and exons are then joined together after transcription to yield the final mRNA for protein translation. An open reading frame may be a continuous stretch of codons. In some embodiments, the open reading frame only applies to spliced mRNAs, not genomic DNA, for expression of a protein.

[0051] “Operably linked” as used herein means that expression of a gene is under the control of a promoter with which it is spatially connected. A promoter may be positioned 5' (upstream) or 3' (downstream) of a gene under its control. The distance between the promoter and a gene may be approximately the same as the distance between that promoter and the gene it controls in the gene from which the promoter is derived. As is known in the art, variation in this distance may be accommodated without loss of promoter function. Nucleic acid or amino acid sequences are “operably linked” (or “operatively linked”) when placed into a functional relationship with one another. For instance, a promoter or enhancer is operably linked to a coding sequence if it regulates, or contributes to the modulation of, the transcription of the coding sequence. Operably linked DNA sequences are typically contiguous, and operablyDocket No. 028193-0052-WO01linked amino acid sequences are typically contiguous and in the same reading frame. However, since enhancers generally function when separated from the promoter by up to several kilobases or more and intronic sequences may be of variable lengths, some polynucleotide elements may be operably linked but not contiguous. Similarly, certain amino acid sequences that are non-contiguous in a primary polypeptide sequence may nonetheless be operably linked due to, for example folding of a polypeptide chain. With respect to fusion polypeptides, the terms “operatively linked” and “operably linked” can refer to the fact that each of the components performs the same function in linkage to the other component as it would if it were not so linked.

[0052] A “peptide” or “polypeptide” is a linked sequence of two or more amino acids linked by peptide bonds. The polypeptide can be natural, synthetic, or a modification or combination of natural and synthetic. Peptides and polypeptides include proteins such as binding proteins, receptors, and antibodies. The terms “polypeptide”, “protein,” and “peptide” are used interchangeably herein. “Primary structure” refers to the amino acid sequence of a particular peptide. “Secondary structure” refers to locally ordered, three dimensional structures within a polypeptide. These structures are commonly known as domains, for example, enzymatic domains, extracellular domains, transmembrane domains, pore domains, and cytoplasmic tail domains. “Domains” are portions of a polypeptide that form a compact unit of the polypeptide and are typically 15 to 350 amino acids long. Typical domains are made up of sections of lesser organization such as stretches of beta-sheet and alpha-helices. “Tertiary structure” refers to the complete three-dimensional structure of a polypeptide monomer. “Quaternary structure” refers to the three-dimensional structure formed by the noncovalent association of independent tertiary units. A “motif” is a portion of a polypeptide sequence and includes at least two amino acids. A motif may be 2 to 20, 2 to 15, or 2 to 10 amino acids in length. In some embodiments, a motif includes 3, 4, 5, 6, or 7 sequential amino acids. A domain may be comprised of a series of the same type of motif.

[0053] “Phase transition” or “transition” may refer to the aggregation of a polypeptide, which occurs sharply at a specific temperature.

[0054] “Promoter” as used herein means a synthetic or naturally derived molecule which is capable of conferring, activating or enhancing expression of a nucleic acid in a cell. A promoter may comprise one or more specific transcriptional regulatory sequences to further enhance expression and / or to alter the spatial expression and / or temporal expression of same. ADocket No. 028193-0052-WO01promoter may also comprise distal enhancer or repressor elements, which may be located as much as several thousand base pairs from the start site of transcription. A promoter may be derived from sources including viral, bacterial, fungal, plants, insects, and animals. A promoter may regulate the expression of a gene component constitutively, or differentially with respect to cell, the tissue or organ in which expression occurs or, with respect to the developmental stage at which expression occurs, or in response to external stimuli such as physiological stresses, pathogens, metal ions, or inducing agents.

[0055] The term “recombinant” when used with reference to, for example, a cell, nucleic acid, protein, or vector, indicates that the cell, nucleic acid, protein, or vector, has been modified by the introduction of a heterologous nucleic acid or protein or the alteration of a native nucleic acid or protein, or that the cell is derived from a cell so modified. Thus, for example, recombinant cells express genes that are not found within the native (naturally occurring) form of the cell or express a second copy of a native gene that is otherwise normally or abnormally expressed, under expressed, or not expressed at all.

[0056] “Reporter,” “reporter group,” “label,” and “detectable label” are used interchangeably herein. The reporter is capable of generating a detectable signal. The label can produce a signal that is detectable by visual or instrumental means. A variety of reporter groups can be used, differing in the physical nature of signal transduction (e.g., fluorescence, electrochemical, nuclear magnetic resonance (NMR), and electron paramagnetic resonance (ERR)) and in the chemical nature of the reporter group. Various reporters include signal-producing substances, such as chromagens, fluorescent compounds, chemiluminescent compounds, radioactive compounds, and the like. In some embodiments, the reporter comprises a fluorescent compound. Reporters may include moieties that produce light, e.g., acridinium compounds, and moieties that produce fluorescence, e.g., fluorescein. In some embodiments, the signal from the reporter is a fluorescent signal. The reporter may comprise a fluorophore. Examples of fluorophores include, but are not limited to, acrylodan (6-acryloy 1-2-dimethylaminonaphthalene), badan (6-bromo-acetyl-2-dimethylamino- naphthalene), rhodamine, naphthalene, danzyl aziridine, 4-[ / V-[(2-iodoacetoxy)ethyl]- / V- methylamino]-7-nitrobenz-2-oxa-1,3-diazole ester (IANBDE), 4-[ / V-[(2-iodoacetoxy)ethyl]-A / - methylamino-7-nitrobenz-2-oxa-1,3-diazole (IANBDA), fluorescein, dipyrrometheneboron difluoride (BODIPY), 4-nitrobenzo[c][1,2,5]oxadiazole (NBD), Alexa fluorescent dyes, and derivatives thereof.Fluorescein derivatives may include, for example, 5-fluorescein, 6-carboxyfluorescein, 3'6-Docket No. 028193-0052-WO01carboxyfluorescein, 5(6)-carboxyfluorescein, 6-hexachlorofluorescein, 6-tetrachlorofluorescein, fluorescein, and isothiocyanate.

[0057] “Sample” or “test sample” as used herein can mean any sample in which the presence and / or level of a fusion protein or biologic described herein is to be detected or determined or any sample comprising a fusion protein or biologic or component thereof as detailed herein. Samples may include liquids, solutions, emulsions, or suspensions. In some embodiments, the sample comprises a cell culture supernatant. Samples can be obtained by any means known in the art. The sample can be used directly as obtained from a cell culture or can be pre-treated, such as by filtration, distillation, extraction, concentration, centrifugation, inactivation of interfering components, addition of reagents, and the like, to modify the character of the sample in some manner as discussed herein or otherwise as is known in the art.

[0058] By “specifically binds,” it is generally meant that a polypeptide (e.g., the disclosed fusion protein) binds to a target when it binds to that target more readily than it would bind to a random, unrelated target.

[0059] “Subject” as used herein can mean a mammal that wants or is in need of the herein described fusion proteins or biologies. The subject may be a patient. The subject may be a human or a non-human animal. The subject may be a mammal. The mammal may be a primate or a non-primate. The mammal can be a primate such as a human; a non-primate such as, for example, dog, cat, horse, cow, pig, mouse, rat, camel, llama, goat, rabbit, sheep, hamster, and guinea pig; or non-human primate such as, for example, monkey, chimpanzee, gorilla, orangutan, and gibbon. The subject may be of any age or stage of development, such as, for example, an adult, an adolescent, or an infant.

[0060] “Substantially identical” can mean that a first and second amino acid or polynucleotide sequence are at least 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% over a region of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 200, 300, 400, 500, 600, 700, 800, 900, 1000, or 1100 amino acids or nucleotides, respectively.

[0061] “Transition” or “phase transition” refers to the aggregation of thermally responsive polypeptides, such as the ELP and fusion protein thereof. Phase transition occurs sharply and reversibly at a specific temperature called the lower critical solution temperature (LCST) or the inverse transition temperature Tt. A transition temperature (Tt) is a temperature at which aDocket No. 028193-0052-WO01fusion protein described herein changes from one state to another. States may include, for example, soluble polypeptides, gels, and aggregates of varying sizes and dimensions. A fusion protein described herein may have a transition temperature of heating (Tt-heating) and a transition temperature of cooling (Tt-cooling). Below the Tt, the thermally responsive polypeptide (or a polypeptide comprising a thermally responsive polypeptide) is highly soluble. Upon heating past the transition temperature, the thermally responsive polypeptides hydrophobically collapse and aggregate, forming a separate, gel-like phase. “Inverse transition cycling” refers to a protein purification method for thermally responsive polypeptides (or a polypeptide comprising a thermally responsive polypeptide). The protein purification method may involve the use of thermally responsive polypeptide’s reversible phase transition behavior to cycle the solution through soluble and insoluble phases, thereby removing contaminants.

[0062] “Treatment” or “treating” or “treatment” when referring to protection of a subject from a disease, means suppressing, repressing, reversing, alleviating, ameliorating, or inhibiting the progress of disease, or completely eliminating a disease. A treatment may be either performed in an acute or chronic way. The term also refers to reducing the severity of a disease or symptoms associated with such disease prior to affliction with the disease. Preventing the disease involves administering a composition of the present disclosure to a subject prior to onset of the disease. Suppressing the disease involves administering a composition of the present disclosure to a subject after induction of the disease but before its clinical appearance. Repressing or ameliorating the disease involves administering a composition of the present disclosure to a subject after clinical appearance of the disease.

[0063] “Variant” used herein with respect to a polynucleotide means (i) a portion or fragment of a referenced nucleotide sequence; (ii) the complement of a referenced nucleotide sequence or portion thereof; (iii) a nucleic acid that is substantially identical to a referenced nucleic acid or the complement thereof; or (iv) a nucleic acid that hybridizes under stringent conditions to the referenced nucleic acid, complement thereof, or a sequences substantially identical thereto.

[0064] “Variant” with respect to a peptide or polypeptide that differs in amino acid sequence by the insertion, deletion, or conservative substitution of amino acids, but retain at least one biological activity. Variant may also mean a protein with an amino acid sequence that is substantially identical to a referenced protein with an amino acid sequence that retains at least one biological activity. Representative examples of “biological activity” include the ability to be bound by a specific antibody or polypeptide or to promote an immune response. Variant canDocket No. 028193-0052-WO01mean a functional fragment thereof. Variant can also mean multiple copies of a polypeptide. The multiple copies can be in tandem or separated by a linker. A conservative substitution of an amino acid, for example, replacing an amino acid with a different amino acid of similar properties (for example, hydrophilicity, degree and distribution of charged regions) is recognized in the art as typically involving a minor change. These minor changes may be identified, in part, by considering the hydropathic index of amino acids, as understood in the art (Kyte et al., J. Mol. Biol. 1982, 157, 105-132). The hydropathic index of an amino acid is based on a consideration of its hydrophobicity and charge. It is known in the art that amino acids of similar hydropathic indexes may be substituted and still retain protein function. In one aspect, amino acids having hydropathic indexes of ±1 are substituted. The hydrophilicity of amino acids may also be used to reveal substitutions that would result in proteins retaining biological function. A consideration of the hydrophilicity of amino acids in the context of a peptide permits calculation of the greatest local average hydrophilicity of that peptide. Substitutions may be performed with amino acids having hydrophilicity values within ±1 of each other. Both the hydrophobicity index and the hydrophilicity value of amino acids are influenced by the particular side chain of that amino acid. Consistent with that observation, amino acid substitutions that are compatible with biological function are understood to depend on the relative similarity of the amino acids, and particularly the side chains of those amino acids, as revealed by the hydrophobicity, hydrophilicity, charge, size, and other properties.

[0065] “Vector” as used herein means a nucleic acid sequence containing an origin of replication. A vector may be a viral vector, bacteriophage, bacterial artificial chromosome, or yeast artificial chromosome. A vector may be a DNA or RNA vector. A vector may be a self-replicating extrachromosomal vector or a DNA plasmid. For example, the vector may encode a Biologic-ELP fusion protein.2. Fusion Proteins

[0066] Described herein are fusion proteins comprising a biologic and an elastin-like polypeptide (ELP). The biologic is attached to the ELP, e.g., operably linked or attached. The ELP may be attached to the N-terminus or the C-terminus of the biologic. In some embodiments, the ELP is attached to the C-terminus of the biologic. The ELP may be attached to the C-terminus or the N-terminus of a heavy chain or a light chain of a biologic. The biologic may comprise one ELP. The biologic may comprise more than one ELP. For example, the biologic may comprise 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 ELPs. The biologic may comprise at least 1,Docket No. 028193-0052-WO01at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, or at least 9 ELPs. The biologic may comprise at most 1, at most 2, at most 3, at most 4, at most 5, at most 6, at most 7, at most 8, at most 9, or at most 10 ELPs.

[0067] In some embodiments, the ELP is attached to the C-terminus or the N-terminus of the heavy chain of a biologic. In some embodiments, the ELP is attached to the C-terminus or the N-terminus of the light chain of a biologic.

[0068] In some embodiments, the biologic comprises a plurality of ELPs, with individual ELPs independently attached to the C-terminus or N-terminus of the heavy chain, theC-terminus or N-terminus of the light chain, or a combination thereof. In some embodiments, an antibody comprises more than one ELP, each ELP being independently attached to the C-terminus of a heavy chain of the antibody, a C-terminus of a light chain of the antibody, or a combination thereof. In some embodiments, an ELP is attached to the C-terminus of each heavy chain of an antibody. In some embodiments, an ELP is attached to the C-terminus of each light chain of an antibody. In some embodiments, an ELP is attached to the C-terminus of each heavy chain of an antibody and an ELP is attached to the C-terminus of each light chain of an antibody. In some embodiments, an antibody comprises two ELPs, a first ELP attached to a terminus of a first heavy chain and a second ELP attached to a terminus of a second heavy chain. In some embodiments, an antibody comprises two ELPs, a first ELP attached to a terminus of a first light chain and a second ELP attached to a terminus of a second light chain. In some embodiments, an antibody comprises four ELPs, a first ELP attached to a terminus of a first heavy chain, a second ELP attached to a terminus of a second heavy chain, a third ELP attached to a terminus of a first light chain, and a fourth ELP attached to a terminus of a second light chain. In some embodiments, a Fc-fusion protein comprises one ELP attached to a terminus of a first heavy chain or a second heavy chain. In some embodiments, a Fc-fusion protein comprises two ELPs, a first ELP attached to a terminus of a first heavy chain and a second ELP attached to a terminus of a second heavy chain.

[0069] The fusion proteins described herein may further comprise a secretion domain attached to the N-terminus of the fusion protein. The fusion proteins described herein may also comprise a solubility-enhancing domain. The fusion protein may be capable of undergoing phase separation in a eukaryotic cell culture. The fusion protein can be uniquely expressed in and secreted from eukaryotic cells. Because of this, the fusion protein can have characteristics that distinguish it from the same fusion protein being expressed in bacterial cells. For example,Docket No. 028193-0052-WO01the disclosed fusion protein can have a glycosylation pattern characteristic of the eukaryotic cell it is expressed and secreted from. In some embodiments, the fusion protein comprises a N-linked glycan. In some embodiments, the fusion protein comprises a biologic and an ELP, wherein the fusion protein is expressed and secreted from an eukaryotic cell and has a glycosylation pattern characteristic of the eukaryotic cell. In some embodiments, all Fes are glycosylated. Glycans may be attached to the nitrogen of asparagine residues within amino acid sequence motifs. Matrix-Assisted Laser Desorption / Ionization (MALDI) may be used to measure a change in molecular weight of a polypeptide to determine the presence of post-translational modifications such as glycosylation. In addition, MALDI may be used to determine the presence of glycosylation by measuring the molecular weight of a polypeptide before and after incubation with PNGase F (Peptide-N-Glycosidase F) that cleaves N-linked glycans.

[0070] The fusion protein described herein may exhibit altered binding affinity to neonatal Fc receptor (FcRn) relative to the biologic without the ELP. In some embodiments, the fusion protein exhibits a lower FcRn equilibrium dissociation constant (KD) relative to the biologic without the ELP. In some embodiments, the fusion protein exhibits a higher FcRn KD relative to the biologic without the ELP. In some embodiments, the fusion protein exhibits the same FcRn KD relative to the biologic without the ELP. In some embodiments, the fusion protein exhibits a higher binding affinity for its target relative to the biologic without the ELP. In some embodiments, the fusion protein exhibits the same binding affinity for its target relative to the biologic without the ELP.a. Biologies

[0071] The biologic can be a therapeutic polypeptide. The biologic may be an antibody or a Fc-fusion protein. The biologic may be, but not limited to, a human, a mouse, a hamster, a rat, a goat, or a rabbit antibody or Fc-fusion protein. The antibody may be a monoclonal antibody, an immunoglobulin (Ig; e.g., IgG such as lgG1, lgG2, lgG3, lgG4, lgG2a, IgM, IgA, IgE, and IgD), or a bispecific antibody. The antibody may be any antibody known in the art. Examples of antibodies include, but are not limited to, Trastuzumab (SEQ ID NOs: 110 & 111) and Mouse Antibody-1 (SEQ ID NOs: 113 & 114).

[0072] The Fc-fusion protein may be any Fc-fusion protein known in the art. The Fc-fusion protein may comprise a modified human IgG Fc domain. The Fc-fusion protein may comprise aDocket No. 028193-0052-WO01p35 domain, a p40 domain, an interleukin-12 (IL-12) domain, or a combination thereof.Examples of Fc-fusion proteins include, but are not limited to, Dulaglutide (SEQ ID NO: 112).b. ELPs

[0073] The ELP may be a disordered domain. The disordered domain may comprise any polypeptide that has minimal or no secondary structure as observed by circular dichroism (CD) and has phase transition behavior. The disordered domain may include an amino acid sequence of repeated amino acids, non-repeated amino acids, or a combination thereof.

[0074] The ELP may comprise an amino acid sequence of [VPGXG]m(SEQ ID NO: 1). X may be any amino acid except proline, and m may be an integer greater than or equal to 1. In some embodiments, m may be an integer from 1 to 500. In some embodiments, m may be at least, at most, or exactly 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 1 1, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 105, 110, 115, 120, 125, 130, 135, 140, 145, 150, 155, 160, 165, 170, 175, 180, 185, 190, 195, 200, 205, 210, 215, 220, 225, 230, 235, 240, 245, 250, 255, 260, 265, 270, 275, 280, 285, 290, 295, 300, 305, 310, 315, 320, 325, 330, 335, 340, 345, 350, 355, 360, 365, 370, 375, 380, 385, 390, 395, 400, 405, 410, 415, 420, 425, 430, 435, 440, 445, 450, 455, 460, 465, 470, 475, 480, 485, 490, 495, or 500. In some embodiments, m may be less than 500, less than 400, less than 300, less than 200, or less than 100. In some embodiments, m may be from 1 to 500, from 1 to 400, from 1 to 300, from 1 to 200, from 60 to 180, from 10 to 120, from 20 to 120, from 40 to 120, from 10 to 100, from 20 to 100, from 40 to 100, from 10 to 80, from 20 to 80, or from 40 to 80. In some embodiments, m may be 20, 40, 60, 80, 120, or 180. In some embodiments, m may be from 10 to 120 where the Tt of the ELP fusion is above 2°C from the temperature of the cell culture, e.g., that it is expressed in and secreted from.

[0075] The ELP may have a hydrophobicity effective to promote expression and secretion from the cell without causing precipitation or aggregation. For example, the ELP may have a hydropathicity score of from about 0.5 to about 1.5, about 0.7 to about 1.5, about 0.9 to about 1.5, about 1.1 to about 1.5, about 1.3 to about 1.5, about 0.5 to about 1.3, about 0.5 to about 1.1, about 0.5 to about 0.9, or about 0.5 to about 0.7. In some embodiments, the ELP may have a hydropathicity score of no less than 0.5, no less than 0.6, no less than 0.7, no less than 0.8, no less than 0.9, or no less than 1. In some embodiments, the ELP may have a hydropathicityDocket No. 028193-0052-WO01score of no greater than 1.5, no greater than 1.4, no greater than 1.3, no greater than 1.2, no greater than 1.1, or no greater than 1.

[0076] Amino acid sequences included by the ELP can have a hydropathicity score as described herein. For example, SEQ ID NO: 1 may have a hydropathicity score of from about 0.5 to about 1.5, about 0.7 to about 1.5, about 0.9 to about 1.5, about 1.1 to about 1.5, about 1.3 to about 1.5, about 0.5 to about 1.3, about 0.5 to about 1.1, about 0.5 to about 0.9, or about 0.5 to about 0.7. In some embodiments, SEQ ID NO: 1 may have a hydropathicity score of no less than 0.5, no less than 0.6, no less than 0.7, no less than 0.8, no less than 0.9, or no less than 1. In some embodiments, SEQ ID NO: 1 may have a hydropathicity score of no greater than 1.5, no greater than 1.4, no greater than 1.3, no greater than 1.2, no greater than 1.1, or no greater than 1. Hydropathicity score is measured and denoted as a Kyte-Doolittle value.

[0077] In some embodiments, X may be alanine (Ala), valine (Val), Serine (Ser), Isoleucine (lie), or a mixture thereof. In some embodiments, X may be Val. In some embodiments, X may be Ala. In some embodiments, X may be Ser. In some embodiments, X may be lie. In some embodiments, X may be a mixture of Ala and Val. In some embodiments, X may be a mixture of Ala and Val in a ratio of 1:1, 1:2, 1:3, 1:4, 1:5, 1:6, 1:7, 1:8, 1:9, 1:10, 2:1, 3:1, 4:1, 5:1, 6:1, 7:1, 8:1, 9:1, or 10:1. In some embodiments, X may be a mixture of Ala and Val in a ratio of 1:1 or 1:4. In some embodiments, X may be a mixture of Ala and Val in a ratio from 10:1 to 1:10 (Ala: Val), such as from 5:1 to 1:5 or from 1:1 to 1:4.

[0078] In some embodiments, X may be Ser and lie arranged in alternating blocks, each block comprising from about 1 to about 40 repeats of a single residue. For example, the ELP may include an amino acid sequence of [VPGSG]I-4O-[VPGIG]I-4O, [VPGIG]I-4O-[VPGSG]I-4O, [VPGSG]I-4O-[VPGIG]I.4O-[VPGSG]I.4O-[VPGIG]I.4O, [VPGIG] O-[VPGSG]I.4O-[VPGIG]I.4O-[VPGSG]I-40, [VPGSG]I-80-[VPGIG]I-80, [VPGIG]I-80-[VPGSG]I-80, or [VPGSG]I.4O-[VPGIG]I-4O-[VPGSG]I-4O-[VPGIG]I-4O-[VPGSG]I-4O-[VPGIG]I-4O. The overall ratio of Ser to lie residues in the ELP may be 1:1, 1:2, 1:3, 1:4, 1:5, 1:6, 1:7, 1:8, 1:9, 1:10, 2:1, 3:1, 4:1, 5:1, 6:1, 7:1, 8:1, 9:1, or 10:1. The alternating blocks may be contiguous blocks.

[0079] In some embodiments, the ELP may comprise an amino acid sequence of [VPGVG]40(SEQ ID NO: 2); [VPGAG]40(SEQ ID NO: 3); [VPG[AI: VI]G]40(SEQ ID NO: 4);[VPG[A1:V4]G]40(SEQ ID NO: 5); [VPG[A4: VI]G]40(SEQ ID NO: 6); [VPGAG]80(SEQ ID NO: 11);[VPGVG]80(SEQ ID NO: 12); [VPG[A1:V1]G]80(SEQ ID NO: 14); [VPG[A4: VI]G]80(SEQ ID NO:Docket No. 028193-0052-WO0115); [VPG[AI: V4]G]80(SEQ ID NO: 16); [VPGVG]120(SEQ ID NO: 21); [VPGSG]2o-[VPGIG]2o-[VPGSG]2O-[VPGIG]20(SEQ ID NO: 17); [VPGSG]2O-[VPGIG]2O-[VPGSG]2o-[VPGIG]2o-[VPGIG]2o (SEQ ID NO: 24); [VPGSG]₈₀(SEQ ID NO: 13); or [VPGIG]4o-[VPGSG]4o (SEQ ID NO: 115). In some embodiments, the ELP may comprise an amino acid sequence of [VPGVG]40(SEQ ID NO: 2); [VPGAG]40(SEQ ID NO: 3); [VPG[AI: VI]G]40(SEQ ID NO: 4); [VPG[AI: V4]G]40(SEQ ID NO: 5); [VPG[A4: VI]G]40(SEQ ID NO: 6); [VPGAG]80(SEQ ID NO: 11); [VPGVG]80(SEQ ID NO: 12); [VPG[A1:V1]G]80(SEQ ID NO: 14); [VPG[A4: VI]G]80(SEQ ID NO: 15);[VPG[AI: V4]G]80(SEQ ID NO: 16); or [VPGVG]120(SEQ ID NO: 21). In some embodiments, the ELP may comprise an amino acid sequence of [VPGSG]2o-[VPGIG]2o-[VPGSG]2O-[VPGIG]2O(SEQ ID NO: 17);[VPGSG]2o-[VPGIG]2o-[VPGSG]2o-[VPGIG]2o-[VPGIG]2o (SEQ ID NO: 24); [VPGSG]80(SEQ ID NO: 13); or [VPGIG]40.[VPGSG]40(SEQ ID NO: 115).c. Secretion Domains

[0080] The fusion protein may comprise a secretion domain that can aid in subcellular trafficking. The secretion domain may comprise a mammalian signal peptide or an insect signal peptide. The secretion domain may be cleaved from the fusion protein after or during secretion from the eukaryotic cell. Secretion domains are known in the art, for example secretion domains described in Kober et al., Biotechnology and Bioengineering, 2013; 110(4) and ACS Synth Biol. 2023; 12(8): 2339-2352, each of which is incorporated by reference herein, may be used. Example secretion domains include, but are not limited to an azurocidin (AZ) signal peptide (SEQ ID NO: 28), an interleukin-2 (IL-2) signal peptide (SEQ ID NO: 31), an albumin signal peptide (SEQ ID NO: 32), a CD33 signal peptide (SEQ ID NO: 33), a tissue plasminogen activator (tPA) signal peptide (SEQ ID NO: 34), a human trypsinogen-2 signal peptide (SEQ ID NO: 35), a human chymotrypsinogen signal peptide (SEQ ID NO: 36), a maltose-binding protein (MBP) signal peptide (SEQ ID NO: 37), such as a human IgG 1 Fc Tag (SEQ ID NO: 41), a glutathione S-transferase (GST) Tag (SEQ ID NO: 40), a CD5 signal peptide (SEQ ID NO: 38), an immunoglobulin Kappa light chain signal peptide (SEQ ID NO: 39), a trypsinogen signal peptide (SEQ ID NO: 29), or a preprolactin signal peptide (SEQ ID NO: 30). In some embodiments, the secretion domain is an azurocidin (AZ) signal peptide (SEQ ID NO: 28).d. Secondary Protein Domains

[0081] The fusion protein may comprise a secondary protein domain that can aid the hydrophilicity and / or hydrophobicity balance of the fusion protein. Example secondary proteinDocket No. 028193-0052-WO01domains include, but are not limited to, mCherry, maltose-binding protein (MBP), thioredoxin, N-utilization substance A (NusA), green fluorescent protein (GFP), glutathione S-transferase (GST), an engineered small ubiquitin-like modifier (SUMO) tag (e.g., SUMOstar™), a modified bacterial dehalogenase tag (e.g., HaloTag®), human serum albumin (HSA), and a second Fc-fusion protein or domain thereof (e.g., a p35 domain, a p40 domain, an interleukin-12 (IL-12) domain, or a combination thereof).e. Linkers

[0082] The fusion protein can comprise a linker. The fusion protein may include more than one linker. The linkers may be the same or different from one another. The fusion protein may include at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 11, at least 12, at least 13, at least 14, at least 15, at least 16, at least 17, at least 18, at least 19, at least 20, at least 25, at least 30, at least 35, at least 40, at least 45, at least 50, at least 55, at least 60, at least 65, at least 70, at least 75, at least 80, at least 85, at least 90, at least 95, or at least 100 linkers. The fusion protein may include less than 500, less than 400, less than 300, or less than 200 linkers. The fusion protein may include between 1 and 1000, between 10 and 900, between 10 and 800, or between 5 and 500 linkers.

[0083] The linker may be positioned in between a biologic polypeptide and an ELP polypeptide, in between biologic polypeptides, in between ELP polypeptides, or a combination thereof. Multiple linkers may be positioned adjacent to one another. Multiple linkers may be positioned adjacent to one another and in between the biologic polypeptide and the ELP polypeptide. In some embodiments, the linker attaches the biologic polypeptide to the ELP polypeptide.

[0084] The linker may be a polypeptide of any amino acid sequence and length. The linker may act as a spacer peptide. The linker may occur between polypeptide domains. The linker can comprise charged amino acids. The linker can comprise uncharged amino acids. The linker may be flexible. The linker may comprise at least one glycine and at least one serine. In some embodiments, the linker is a flexible glycine linker, such as a glycine / serine linker. An example linker includes, but is not limited to, [GGGGS]i-s (SEQ ID NO: 25). In some embodiments, the linker is GGGGSGGGGSG (SEQ ID NO: 26).f. Transition PropertiesDocket No. 028193-0052-WO01

[0085] The fusion protein may demonstrate phase transition behavior by changing solubility and aggregate formation with temperature. The phase transition behavior of the fusion protein may derive from the phase transition behavior of the ELP domains of the fusion protein. The phase transition may be reversible, although the specific temperature of dissolution may be the same or different from the specific temperature of aggregation. In some embodiments, the fusion protein may be soluble below a lower critical solution temperature (LCST). LCST is the temperature below which the polypeptide is miscible.

[0086] A transition temperature (Tt) is a temperature at which the fusion protein changes from one state to another. States may include, for example, soluble polypeptides, gels, and aggregates of varying sizes and dimensions. The fusion protein may have a transition temperature of heating (Tt-heating) and a transition temperature of cooling (Tt-cooling). In some embodiments, the Tt-heating may be concentration-dependent. In some embodiments, the Tt-cooling may be concentration-independent. Below the transition temperature (LCST or Tt), the fusion protein may be highly soluble. Upon heating above the transition temperature, the fusion protein may hydrophobically collapse and aggregate, forming a separate phase.

[0087] The ELP and / or fusion protein thereof may have a transition temperature (Tt) that is at least 1 °C higher, at least 2°C higher, at least 3°C higher, at least 4°C higher, at least 5°C higher, at least 10°C higher, at least 15°C higher, or at least 20°C higher than a temperature that the at least one eukaryotic cell is cultured at. The ELP and / or fusion protein may have a Tt that is at most 2°C higher, at most 3°C higher, at most 4°C higher, at most 5°C higher, at most 6°C, at most 10°C, at most 15°C, or at most 20°C higher than a temperature that the at least one eukaryotic cell is cultured at. The ELP and / or fusion protein may have a Tt that is from about 1°C to about 20°C, about 1°C to about 15°, about 2°C to about 20°C, about 5°C to about 20°C, about 1 °C to about 6°C, about 2°C to about 6°C, about 3°C to about 6°C, about 4°C to about 6°C, about 5°C to about 6°C, about 1 °C to about 5°C, about 1 °C to about 4°C, about 1 °C to about 3°C, or about 1°C to about 2°C higher than a temperature that the at least one eukaryotic cell is cultured at.

[0088] The ELP and fusion protein thereof may phase transition at a variety of temperatures. The ELP and / or fusion protein thereof may have a Tt from about 0°C to about 100°C, from about 10°C to about 50°C, or from about 20°C to about 42°C. In some embodiments, the ELP and / or fusion protein may have a Tt of from about 34°C to about 65°C, about 38°C to about 65°C, about 42°C to about 65°C, about 46°C to about 65°C, about 50°C toDocket No. 028193-0052-WO01about 65°C, about 54°C to about 65°C, about 58°C to about 65°C, about 62°C to about 65°C, about 34°C to about 60°C, about 34°C to about 55°C, about 34°C to about 50°C, about 34°C to about 45°C, about 34°C to about 40°C, or about 34°C to about 35°C. In some embodiments, the ELP and / or fusion protein thereof may have Tt of no less than 34°C, no less than 35°C, no less than 36°C, no less than 37°C, no less than 38°C, no less than 39°C, no less than 40°C, no less than 45°C, or no less than 50°C. In some embodiments, the ELP and / or fusion protein thereof may have Tt of no greater than 34°C, no greater than 35°C, no greater than 36°C, no greater than 37°C, no greater than 38°C, no greater than 39°C, no greater than 40°C, no greater than 45°C, or no greater than 50°C.

[0089] The Tt-heating and Tt-cooling may be identical. Temperatures may be “identical” when the temperatures are within 2.0°C, 1,0°C, 0.5°C, or 0.1 °C of each other. In some embodiments, the Tt-heating is greater than the Tt-cooling. In embodiments where the fusion protein has a Tt-heating greater than the Tt-cooling, the difference between the two transition temperatures may be referred to as a hysteresis. In some embodiments, the fusion protein has a hysteresis of about 5°C to about 70°C, such as about 5°C to about 60°C or about 10°C to about 50°C.

[0090] The phase transition behavior of the fusion protein may be utilized in purification of the fusion protein according to a method referred to as “inverse transition cycling,” in which the fusion protein’s reversible phase transition behavior is used to cycle the solution through soluble and insoluble phases, thereby removing contaminants. Phase transition may also be triggered using kosmotropic salts, such as, for example, ammonium sulfate or sodium chloride. The kosmotropic salt may be added to a solution comprising the fusion protein, with the kosmotropic salt being added until the fusion protein forms aggregates or is precipitated out of solution. The aggregates may be pelleted by centrifugation and resuspended in a second solution or buffer. Aggregates of the fusion protein may re-solubilize into solution once cooled below their Tt or when the kosmotropic salt is removed from the solution. In some embodiments, the fusion protein is purified without any chromatographic purification. In some embodiments, the fusion protein is generated recombinantly and purified from eukaryotic culture, such as, for example, from a mammalian cell or insect cell.g. Example Fusion ProteinsDocket No. 028193-0052-WO01

[0091] In some embodiments, the fusion protein comprises a formula of biologic-[VPGXG]10-120 (SEQ ID NO: 1), wherein the ELP and / or SEQ ID NO: 1 has a hydropathicity score of from about 0.5 to about 1.5, and the biologic is a biologic as disclosed herein.

[0092] In some embodiments, the fusion protein comprises a formula of biologic-[VPGXG]10-120 (SEQ ID NO: 1), wherein X is alanine (Ala), valine (Val), Serine (Ser), Isoleucine (lie), or a mixture thereof, and the biologic is a biologic as disclosed herein.3. Polynucleotides

[0093] Further provided are polynucleotides encoding the fusion proteins detailed herein. A vector may include the polynucleotide encoding the fusion proteins detailed herein. To obtain expression of a polypeptide, one may subclone the polynucleotide encoding the polypeptide into an expression vector that contains a promoter to direct transcription, a transcription / translation terminator, and if for a nucleic acid encoding a protein, a ribosome binding site for translational initiation. An example of a vector is pcDNA5. Suitable eukaryotic promoters, such as mammalian and insect promoters, are well known in the art. Further provided is a host cell transformed or transfected with an expression vector comprising a polynucleotide encoding a fusion protein as detailed herein. Eukaryotic expression systems for expressing the fusion protein are available in, for example, human, mouse, hamster, rat, goat, rabbit, yeast, and insects. Kits for such expression systems are commercially available.

[0094] The fusion protein may be expressed recombinantly in a host cell according to one of skill in the art. The fusion protein may be purified by any means known to one of skill in the art. For example, the fusion protein may be purified using chromatography, such as liquid chromatography, size exclusion chromatography, affinity chromatography, and a combination thereof. In some embodiments, the fusion protein is purified without chromatography. In some embodiments, the fusion protein is purified using inverse transition cycling.4. Compositions

[0095] Further provided herein are compositions comprising a fusion protein described herein. The composition may comprise a fusion protein described herein and a cell culture media. Cell culture media are well known in the art and vary depending on the type of eukaryotic cell used. For example, Gibco™ Ham’s F-12 or Dulbecco’s Modified Eagle’s Medium (DM EM) are commonly used for mammalian cell cultures.Docket No. 028193-0052-WO01

[0096] The composition may comprise a fusion protein described herein and a buffer.Suitable buffers are well known in the art. The buffer may be a low salt buffer. The low salt buffer may be phosphate-buffered saline (PBS), Dulbecco’s Phosphate- Buffered Saline (DPBS), citrate buffer, 4-(2-hydroxyethyl)-1 -piperazineethanesulfonic acid (HEPES) buffer, tris(hydroxymethyl)aminomethane (Tris) buffer, acetate buffer, histidine buffer, succinate buffer, or the like.

[0097] Further provided herein are pharmaceutical compositions comprising the fusion protein described herein. In some embodiments, the pharmaceutical composition may comprise about 1 ng to about 10 mg of the fusion protein. The fusion protein may be formulated into a pharmaceutical composition in accordance with standard techniques well known to those skilled in the pharmaceutical art. The pharmaceutical compositions can be formulated according to the mode of administration to be used. In cases where pharmaceutical compositions are injectable pharmaceutical compositions, they are sterile, pyrogen free, and particulate free. An isotonic formulation is preferably used. Generally, additives for isotonicity may include sodium chloride, dextrose, mannitol, sorbitol, and lactose. In some cases, isotonic solutions such as phosphate buffered saline are preferred. Stabilizers include gelatin and albumin. The composition may further comprise a pharmaceutically acceptable excipient. The pharmaceutically acceptable excipient may be functional molecules as vehicles, adjuvants, carriers, or diluents. The term “pharmaceutically acceptable carrier,” may be a non-toxic, inert solid, semi-solid or liquid filler, diluent, encapsulating material or formulation auxiliary of any type. Pharmaceutically acceptable carriers include, for example, diluents, lubricants, binders, disintegrants, colorants, flavors, sweeteners, antioxidants, preservatives, glidants, solvents, suspending agents, wetting agents, surfactants, emollients, propellants, humectants, powders, pH adjusting agents, and combinations thereof.5. Administration

[0098] The polynucleotides as detailed herein, or at least one component thereof, may be administered or delivered to a cell. Methods of introducing a nucleic acid into a host cell are known in the art, and any known method can be used to introduce a nucleic acid (e.g., an expression construct) into a cell. Suitable methods include, for example, viral or bacteriophage infection, transfection, conjugation, protoplast fusion, polycation or lipid:nucleic acid conjugates, lipofection, electroporation, nucleofection, immunoliposomes, calcium phosphate precipitation, polyethyleneimine (PEI)-mediated transfection, DEAE-dextran mediated transfection, liposome-Docket No. 028193-0052-WO01mediated transfection, particle gun technology, calcium phosphate precipitation, direct micro injection, nanoparticle-mediated nucleic acid delivery, and the like. The polynucleotide or a composition comprising the same, may be electroporated using BioRad Gene Pulser Xcell or Amaxa Nucleofector IIb devices or other electroporation device. Several different buffers may be used, including BioRad electroporation solution, Sigma phosphate-buffered saline product #D8537 (PBS), Invitrogen OptiMEM I (OM), or Amaxa Nucleofector solution V (N. V.).Transfections may include a transfection reagent, such as Lipofectamine 2000.

[0099] The fusion protein as detailed herein, or at least one component thereof, or the pharmaceutical compositions comprising the same, may be administered to a subject. Such compositions can be administered in dosages and by techniques well known to those skilled in the medical arts taking into consideration such factors as the age, sex, weight, and condition of the particular subject, and the route of administration.[000100] The fusion proteins, or at least one component thereof, or compositions comprising the same, can be administered prophylactically or therapeutically. In prophylactic administration, the fusion protein can be administered in an amount sufficient to induce a response. In therapeutic applications, the fusion protein is administered to a subject in need thereof in an amount sufficient to elicit a therapeutic effect. An amount adequate to accomplish this is defined as “therapeutically effective dose.” Amounts effective for this use will depend on, e.g., the particular composition of the fusion protein regimen administered, the manner of administration, the stage, and severity of the disease, the general state of health of the patient, and the judgment of the prescribing physician.[000101] The fusion protein can be administered by methods well known in the art as described in Donnelly et al. Ann. Rev. Immunol. 1997, 75, 617-648; U. S. Patent No. 5,580,859; U. S. Patent No. 5,703,055; and U. S. Patent No. 5,679,647, the contents of each of which are incorporated herein by reference in their entirety. The fusion protein can be complexed to particles or beads that can be administered to an individual, for example, using a vaccine gun. One skilled in the art would know that the choice of a pharmaceutically acceptable carrier, including a physiologically acceptable compound, depends, for example, on the route of administration.[000102] The presently disclosed fusion protein, or at least one component thereof, or compositions comprising the same, may be administered to a subject by different routesDocket No. 028193-0052-WO01including orally, parenterally, sublingually, transdermally, rectally, transmucosally, topically, intranasal, intravaginal, via inhalation, via buccal administration, intrapleurally, intravenous, intraarterial, intraperitoneal, subcutaneous, intradermally, epidermally, intramuscular, intranasal, intrathecal, intracranial, and intraarticular or combinations thereof. For veterinary use, the fusion proteins or compositions comprising the same may be administered as a suitably acceptable formulation in accordance with normal veterinary practice. The veterinarian may readily determine the dosing regimen and route of administration that is most appropriate for a particular animal. The fusion proteins or compositions comprising the same may be administered by traditional syringes, needleless injection devices, or other physical methods.[000103] The fusion protein can be in a liquid preparation such as a suspension, syrup, or elixir. The fusion protein can be incorporated into liposomes, microspheres, or other polymer matrices (such as by a method described in U. S. Patent No. 5,703,055; Gregoriadis, Liposome Technology, Vols. I to III (2nd ed. 1993), the contents of which are incorporated herein by reference in their entirety). Liposomes can consist of phospholipids or other lipids, and can be nontoxic, physiologically acceptable, and metabolizable carriers that are relatively simple to make and administer.[000104] The fusion protein may be used as a vaccine. The vaccine can be administered via electroporation, such as by a method described in U. S. Patent No. 7,664,545, which is incorporated herein by reference. The electroporation can be by a method or apparatus described in U. S. Patent No. 6,302,874; U. S. Patent No. 5,676,646; U. S. Patent No. 6,241,701; U. S. Patent No. 6,233,482; U. S. Patent No. 6,216,034; U. S. Patent No. 6,208,893; U. S. Patent No. 6,192,270; U. S. Patent No. 6,181,964; U. S. Patent No. 6,150,148; U. S. Patent No.6,120,493; U. S. Patent No. 6,096,020; U. S. Patent No. 6,068,650; and U. S. Patent No.5,702,359, the contents of each of which are incorporated herein by reference in their entirety. The electroporation can be carried out via a minimally invasive device.[000105] The fusion protein may be administered in a controlled release formulation. The fusion protein may comprise one or more thermally responsive polypeptides (e.g., ELP), the thermally responsive polypeptide having a transition temperature such that the fusion protein remains soluble prior to administration and such that the fusion protein transitions upon administration to a gel-like depot in the subject. For example, the fusion protein may comprise one or more thermally responsive polypeptides, the thermally responsive polypeptide having a transition temperature between room temperature (about 25 °C) and body temperature (aboutDocket No. 028193-0052-WO0137 °C), whereby the fusion protein can be administered to form a depot. As used herein, “depot” refers to a gel-like composition comprising a fusion protein that releases the fusion protein overtime. The fusion protein can be injected subcutaneously or intratumorally to form a depot (coacervate). The depot may provide controlled (slow) release of the fusion protein. The depot may provide slow release of the fusion protein into circulation or a tumor, for example. In some embodiments, the fusion protein may be released from the depot over a period of at least about 1 day, at least about 2 days, at least about 3 days, at least about 4 days, at least about 5 days, at least about 6 days, at least about 7 days, at least about 1 week, at least about 1.5 weeks, at least about 2 weeks, at least about 2.5 weeks, at least about 3.5 weeks, at least about 4 weeks, or at least about 1 month.6. Kits[000106] Provided herein is a kit, which may be used to purify or administer a fusion protein. The kit comprises one or more fusion proteins or a composition comprising the same, for purifying or administering a fusion protein, as described herein, and instructions for purifying a fusion protein by reversible phase separation or administering a fusion protein by means described herein. In some embodiments, the kit comprises at least one fusion protein having an amino acid sequence of any one of SEQ ID NOs: 1-24, 42-109, and 115, a complement thereof, a variant thereof, or fragment thereof, and instructions for using the fusion protein. The kit may also comprise a buffer suitable for inducing or reversing a phase transition of the ELP.[000107] Instructions included in kits may be affixed to packaging material or may be included as a package insert. While the instructions are typically written on printed materials they are not limited to such. Any medium capable of storing such instructions and communicating them to an end user is contemplated by this disclosure. Such media include, but are not limited to, electronic storage media (e.g., magnetic discs, tapes, cartridges, chips), optical media (e.g., CD ROM), and the like. As used herein, the term “instructions” may include the address of an internet site that provides the instructions.7. Methodsa. Methods of Producing a Biologic-Based Fusion Protein[000108] Provided herein are methods of producing a biologic-based fusion protein. The methods may include engineering a eukaryotic cell to express a fusion protein as describedDocket No. 028193-0052-WO01herein. The methods may further include culturing the eukaryotic cell under conditions permitting secretion of the fusion protein into a culture supernatant; and recovering the fusion protein from the culture supernatant. In some embodiments, the fusion protein is recovered without chromatography.[000109] The eukaryotic cell can be a mammalian cell or an insect cell. Example mammalian cells include, but are not limited to, a human cell, a mouse cell, a hamster cell, a rat cell, a goat cell, or a rabbit cell. Cell lines for these types of cells are well known within the art. Example cell lines include, but are not limited to, ExpiCHO cells, CHO cells, Expi293 cells, HEK293 cells, HeLa, Vero, Jurkat, primary cells isolated directly from tissue, pluripotent or multipotent stem cells, immortalized cells, Sf9 cells, Sf21 cells, S2 cells, High Five cells, Bm5 cells, Tn368 cells, and the like.[000110] Engineering a eukaryotic cell to express a fusion protein may comprise transfecting the eukaryotic cell with a plasmid or vector comprising a polynucleotide encoding a fusion protein as described herein. Then, culturing the eukaryotic cell in a cell culture media suitable for the cell type being used for a period time such as for at least about 1 day, at least about 2 days, at least about 3 days, at least about 4 days, at least about 5 days, at least about 6 days, at least about 7 days, at least about 1 week, at least about 1.5 weeks, at least about 2 weeks, at least about 2.5 weeks, at least about 3.5 weeks, at least about 4 weeks, or at least about 1 month.[000111] The eukaryotic cell may be cultured at a temperature suitable for the cell line being used. In some embodiments, the at least one eukaryotic cell may be cultured at a temperature of from about 30°C to about 37°C, about 31°C to about 37°C, about 32°C to about 37°C, about 33°C to about 37°C, about 34°C to about 37°C, about 35°C to about 37°C, about 36°C to about 37°C, about 32°C to about 36°C, about 32°C to about 35°C, about 32°C to about 34°C, or about 32°C to about 33°C. In some embodiments, the at least one eukaryotic cell may be cultured at a temperature of no less than 30°C, no less than 31 °C, no less than 32°C, or no less than 33°C. In some embodiments, the at least one eukaryotic cell may be cultured at a temperature of no greater than 37°C, no greater than 36°C, no greater than 35°C, or no greater than 34°C.[000112] Recovering the fusion protein from the culture supernatant may comprise harvesting the cell culture, centrifuging the cell culture at a speed and for a period of time, and collecting the cell culture supernatant. The centrifugation speed may be at least 1000g, at least 2000g, atDocket No. 028193-0052-WO01least 3000g, at least 4000g, at least 5000g, at least 6000g, or at least 7000g. The period of time for which the cell culture is centrifuged may be at least 1 minute, at least 5 minutes, at least 10 minutes, at least 15 minutes, at least 20 minutes, or at least 25 minutes. The cell culture supernatant can then be dialyzed against a buffer such as PBS for at least 10 hours, at least 15 hours, at least 20 hours, at least 24 hours, at least 30 hours, or at least 35 hours. Then, the dialyzed supernatant may be filtered through an about 0.1 micron, about 0.2 micron, about 0.3 micron, about 0.4 micron, about 0.5 micron, about 0.6 micron, about 0.7 micron, or about 0.8 micron filter and purified as described herein.b. Methods of Purifying a Fusion Protein[000113] Provided herein are methods of purifying a fusion protein. The methods may include providing a culture supernatant comprising a fusion protein described herein; inducing a phase transition of the ELP to form an insoluble phase; isolating the insoluble phase from the supernatant; and resolubilizing the fusion protein. In some embodiments, the biologic is purified without chromatography.[000114] Inducing a phase transition of the ELP may comprise increasing the temperature of the supernatant above a transition temperature (Tt) of the ELP.[000115] Inducing a phase transition of the ELP may comprise adding from about 0.05M to about 5M, about 0.1M to about 5M, about 0.5M to about 5M, about 1M to about 5M, about 1.5M to about 5M, about 2M to about 5M, about 3M to about 5M, about 4M to about 5M, about 0.05M to about 4M, about 0.05M to about 3M, about 0.05M to about 2M, about 0.05M to about 1,5M, about 0.05M to about 1M, or about 0.05M to about 0.1M salt to the supernatant. In some embodiments, the salt may be added to the supernatant at no less than 0.05M, no less than 0.1M, no less than 0.5M, no less 0.6M, no less than 0.7M, no less 0.8M, no less 0.9M, or no less than 1 M. In some embodiments, the salt may be added to the supernatant at no greater than 5M, no greater than 4M, no greater than 3M, no greater than 2M, no greater 1.9M, no greater than 1,8M, no greater 1,7M, no greater than 1,6M, or no greater than 1,5M. The salt may be a kosmotropic salt. In some embodiments, the salt may be sodium chloride, ammonium sulfate, sodium citrate, sodium sulfate, magnesium sulfate, potassium sulfate, sodium phosphates, lithium chloride, sodium acetate, or sodium carbonate.[000116] Isolating the insoluble phase from the supernatant may comprise centrifugation or tangential flow filtration.Docket No. 028193-0052-WO01[000117] Resolubilizing the fusion protein may comprise adding a low salt buffer and removing insoluble contaminants using centrifugation or tangential flow filtration. The low salt buffer may be phosphate-buffered saline (PBS), Dulbecco’s Phosphate- Buffered Saline (DPBS), citrate buffer, 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (HEPES) buffer, tris(hydroxymethyl)aminomethane (Tris) buffer, acetate buffer, histidine buffer, succinate buffer, or the like.c. Methods of Treating a Disease or Disorder[000118] Provided herein are methods of treating a disease or disorder. The methods may include administering to a subject a therapeutically effective amount of a fusion protein described herein. The fusion protein may exhibit an extended serum half-life relative to a biologic without the ELP. The fusion protein may be administered subcutaneously and form a depot at the administration site.[000119] The disease or disorder may be any disease or disorder known to be targeted or treated by a biologic disclosed herein. In some embodiments, the disease is a disease associated with a target of the biologic. Examples of diseases and disorders include, but are not limited to cancer, metabolic disease, autoimmune disease, cardiovascular disease, and orthopedic disorders.[000120] Metabolic disease may occur when abnormal chemical reactions in the body alter the normal metabolic process. Metabolic diseases may include, for example, insulin resistance, non-alcoholic fatty liver diseases, type 2 diabetes, insulin resistance diseases, cardiovascular diseases, arteriosclerosis, lipid-related metabolic disorders, hyperglycemia, hyperinsulinemia, hyperlipidemia, and glucose metabolic disorders.[000121] Autoimmune diseases arise from an abnormal immune response of the body against substances and tissues normally present in the body. Autoimmune diseases may include, but are not limited to, lupus, rheumatoid arthritis, multiple sclerosis, insulin dependent diabetes mellitus, myasthenia gravis, Grave's disease, autoimmune hemolytic anemia, autoimmune thrombocytopenia purpura, Goodpasture's syndrome, pemphigus vulgaris, acute rheumatic fever, post-streptococcal glomerulonephritis, polyarteritis nodosa, myocarditis, psoriasis, Celiac disease, Crohn's disease, ulcerative colitis, and fibromyalgia.Docket No. 028193-0052-WO01[000122] Cardiovascular disease is a class of diseases that involve the heart or blood vessels. Cardiovascular diseases may include, for example, coronary artery diseases (CAD) such as angina and myocardial infarction (heart attack), stroke, hypertensive heart disease, rheumatic heart disease, cardiomyopathy, heart arrhythmia, congenital heart disease, valvular heart disease, carditis, aortic aneurysms, peripheral artery disease, and venous thrombosis.[000123] Orthopedic disorders or musculoskeletal disorders are injuries or pain in the body’s joints, ligaments, muscles, nerves, tendons, and structures that support limbs, neck, and back. Orthopedic disorders may include degenerative diseases and inflammatory conditions that cause pain and impair normal activities. Orthopedic disorders may include, for example, carpal tunnel syndrome, epicondylitis, and tendinitis.[000124] Cancers may include, but are not limited to, breast cancer, colorectal cancer, colon cancer, lung cancer, prostate cancer, testicular cancer, brain cancer, skin cancer, rectal cancer, gastric cancer, esophageal cancer, sarcomas, tracheal cancer, head and neck cancer, pancreatic cancer, liver cancer, ovarian cancer, lymphoid cancer, cervical cancer, vulvar cancer, melanoma, mesothelioma, renal cancer, bladder cancer, thyroid cancer, bone cancers, carcinomas, sarcomas, and soft tissue cancers.8. Examples[000125] The foregoing may be better understood by reference to the following examples, which are presented for purposes of illustration and are not intended to limit the scope of the disclosure. The present disclosure has multiple aspects and embodiments, illustrated by the appended non-limiting examples.Example 1Benefits of Mammalian Cell Secretion of Biologic-ELP Fusions[000126] To date, mAb-ELP and FcfP-ELP fusions have never been expressed and secreted from mammalian cells, as ELPs could only be expressed and purified from E. coli and had to be chemically or enzymatically conjugated to mAbs or FcfPs expressed in eukaryotic (mammalian-and insect-derived cells). The need to express ELPs in bacterial systems greatly increases the complexity and cost to develop and produce mAb-ELPs and FcfP-ELPs. Furthermore, use of E. co / / -generated ELPs introduces the potential risk of endotoxin contamination in the final product, which can cause adverse effects during the administration of therapeutics.Docket No. 028193-0052-WO01[000127] The mAbs and FcfPs fused to ELPs in mammalian cells have several properties, such as: mab-ELPs and FcfP-ELPs present concentration dependent phase-transition behavior that is derived from the ELP but impacted by the fusion protein; mab-ELPs and FcfP-ELPs can be purified through reversible phase separation driven by the ELP tag; mab-ELP and FcfP-ELPs retain binding to their original targets but can be marginally impacted; ELP fusions can be expressed in different mammalian expression systems such as HEK and CHO cells; ELP heavy chain fusions preserve the protein’s original chemical stability; mab-ELP and FcfP-ELP have varying affinities (ELP architecture dependent) for the neonatal Fc receptor (FcRn) that controls mAb and FcfP cellular recycling and is the key to the long half-lives of mAbs and FcfP’s; and mab-ELP and FcfP-ELP have varying affinities for other transmembrane glycoproteins — such as the Fc gamma receptor (FcyR) — involved in mAb and FcfP cellular effector functions, and the affinity is also dependent on the ELP architecture.[000128] The development of mAb-ELP and FcfP-ELP fusions that can be expressed in mammalian cells: eliminates the need for expensive Protein A / G chromatography equipment that is typically used for mAb and FcfP purification, as the ELP can drive phase separation of the fusion partner, enabling mAb-ELP and FcfP-ELP purification through centrifugation and / or tangential flow filtration (TFF); eliminates the need for a harsh, low pH elution step during chromatography-based purification, which can improve final product stability and activity, especially for highly sensitive proteins; eliminates the need to express ELPs in bacterial systems to create ELP fusions with proteins that can only be expressed in mammalian cell systems (e.g. therapeutic antibodies) which would require separate purification and subsequent conjugation to mAbs and FcfPs that are expressed in mammalian- or insect-derived cells to generate mAb-ELPs and FcfPs-ELPs; enables the design of mAb-ELPs and FcfP-ELPs that can phase-transition below a pre-determined temperature so that the mAb-ELP or FcfP-ELP can be injected s.c., intramuscularly (i.m.), or intratumorally (i.t.) as a solution, forming a coacervate at the injection site for sustained release of the fusion for local or systemic delivery; enables the design of mAb-ELPs and FcfP-ELPs that phase-transition above body temperature, so that the mAb-ELP or FcfP-ELP can circulate as a soluble conjugate in systemic circulation; enables the design of mAb-ELPs and FcfP-ELP with unique reactive residues on the ELP(s) to which one or more copies of drugs, radionuclides, fluorophores or imaging agents can be conjugated; enables the design of mAb-ELPs and FcfP-ELPs with controlled charge and limited interaction with hyaluronic acid and other elements from the extracellular matrix for improved diffusion and absorption in the subcutaneous space; enables the design of mAb-ELP- and FcfP-ELP-basedDocket No. 028193-0052-WO01self-assembling nanostructures for drug delivery applications such as micelles and vesicles that phase-transition below a pre-determined temperature and sequester the mAb and FcfP in the interior or present the mAb and FcfP on the exterior of the nanoparticle; minimizes supplies, beyond mammalian cell culture equipment, the only required equipment consists of centrifuges or a TFF setup, common and inexpensive salts and buffers, standard refrigerators, and standard labware to purify mAb-ELPs and FcfP-ELPs; minimizes training, scientists using this technology only need to know how to grow mammalian cells in bioreactors and execute standard lab techniques no knowledge of bacterial cell culture and protein purification techniques or advanced protein chromatography techniques is needed to isolate mAb-ELPs and FcfP-ELPs.Example 2Development of ELPs secreted by mammalian cells[000129] Elastin-like polypeptides (ELPs) have been expressed and purified from E. coli for over two decades. However, their highly repetitive amino acid sequence, and thus their highly repetitive DNA sequence, coupled with their hydrophobicity and propensity to phase separate (clump together) inside cells, has hampered their expression and secretion from mammalian cells. We note that ELPs have been previously expressed within mammalian cells, but their secretion was not demonstrated (Pastuszka et al., 2012, Shi et al., 2014, Pastuszka et al., 2014, Li et al., 2018), especially when fused to mAbs and FcfPs. Previous attempts to express and secrete ELPs in mammalian cells have consistently failed. From the perspective of highly repetitive amino acid sequences with various degrees of hydrophobicity and trafficking of the ELPs and ELP-fusions through the secretory pathway, we believe these failures are due to the fact that in E. coli, the protein is translated by ribosomes in the cytoplasm where they remain until the protein is harvested by lysing the cell. In mammalian systems, proteins are translated by ribosomes attached to the endoplasmic reticulum (rough endoplasmic reticulum), where they are folded and transported to the Golgi apparatus through cargo vesicles. In the Golgi apparatus they are further processed, sorted and packaged into new vesicles. Proteins tagged for secretion are then transported to the plasma membrane for secretion / exocytosis. ELP’s tendency to phase separate inside cells at standard cell culture temperatures (32°C - 37°C), especially for longer and more hydrophobic ELPs with transition temperature below 30°C -which are also the ones with greater biotechnology applications and traditionally investigated in material science- can hamper protein production and secretion pathways. To overcome theseDocket No. 028193-0052-WO01issues, we designed mAb-ELP and FcfP-ELP fusions with varying degrees of valine and alanine guest residues with 40, 80, and 120 repeats of the VPGXG sequence where “X” is the guest residue. We also evaluated the impact of fusing the ELPs to the heavy, light and heavy and light chains of antibodies. To ensure expressed proteins were tagged for secretion we added the azurocidin signal peptide to the N-terminus of all constructs.Example 3Description of mAb-ELP and FcfP-ELP expression and characterization[000130] To demonstrate the ability to express different ELPs fused to mAbs and FcfP proteins, we designed a set of ELPs with DNA sequences for mammalian cell expression fused to mAb heavy and / or light chain C-terminus.[000131] First, we generated separate pcDNA5 plasmids containing Trastuzumab’s heavy and light chains with ELPs where the VPGXG guest residue “X” was valine or arginine in different ratios fused to the C-terminus of Trastuzumab heavy (HC) and / or light chains (LC). Plasmids for heavy and light chain with DNA for mammalian cell expression were transfected into Expi293 cells following the manufacturer instructions. After 7 days of culture, we harvested the cell culture which was clarified through centrifugation at 5000g for 15 minutes. The cell culture supernatant was then dialyzed against 1xPBS for 24 hours, filtered through a 0.2 micron filter and purified with a Cytiva Protein A chromatography column following the manufacturer recommendations. Following purification, we ran an SDS-Page gel under non-reducing and reducing conditions as shown in FIGS. 1A-B, where we were able to confirm the successful expression and purification of several antibody (Trastuzumab) ELP fusions.[000132] Following this initial assessment of expression and secretion of a 40-mer library, where all ELPs presented 40 repeats of the VPGXG amino acid sequence with different ratios of Valine and Alanine - 100% valine V(40) (SEQ ID NO: 2); 75% valine and 25% alanine, V4A1(40) (SEQ ID NO: 5); 50% valine and 50% alanine, V1A1(40) (SEQ ID NO: 4); 25% valine and 75% alanine, V1A4(40) (SEQ ID NO: 6); 100% alanine, A(40) (SEQ ID NO: 3) - we calculated the expression yields in comparison to the parental Trastuzumab. As can be noted in FIG. 2, yields were dependent on fusions architecture and increase with the decrease of ELP hydrophobicity. In general, fusions to the HC also presented higher yields than those to the LC and HC and LC.Docket No. 028193-0052-WO01[000133] Following this initial evaluation, we sought to investigate the impact the different ELP architectures and fusions sites (HC, LC and HC+LC) can have on the potency of the Trastuzumab parental antibody. To this end, half-maximal effective concentration (EC50) assays were performed in with Chemiluminescent ELISA. 100µL per well of a 1µg / mL solution of HER2 in bicarbonate buffer was incubate in MaxiSorp plates overnight. Next, plates were washed 3 times with IxTBST buffer and blocked with a 1XTBST solution with 2% BSA for 1 hour. After 3 rounds of IxTBST wash, microwells were incubated with 100pL of trastuzumab or trastuzumab-ELP solutions with various concentrations ranging from 0.03 nM to 100 nM in 1xPBS in triplicate. Following 3 rounds of washing with IxTBST, wells were incubated for 1-hour with an anti-human IgG antibody conjugated to HRP and washed with IxTBST 3 more times. Finally, wells were incubated with a supersignal chemiluminescent substrate and imaged at 425nm. As can be seen in FIG. 3, there were small affinity shifts in the Trastuzumab-ELP fusions when compared to the parental antibody. These results indicate that antibody-ELP fusions may increase binding of the antibody, which we hypothesize is caused by the formation of nanostructures of Trastuzumab-ELPs and increase in binding valency of the system.[000134] Following these initial potency studies, we sought to investigate if the fusion of ELPs to antibodies, in this case Trastuzumab’s HC, can affect the antibody chemical stability. To this end, we performed guanidine-HCI denaturation assays in a subset of Trastuzumab-ELPs versus the unmodified Trastuzumab as shown in FIGS. 4A-B, where trastuzumab or trastuzumab-ELP at a final concentration of 0.5 µM was denatured overnight at room temperature in guanidine-HCI pH = 7.4 at concentration varying from 0M to 5.5M in 0.5M increments. Assays were done in triplicate in clear bottom plates. Intrinsic tryptophan fluorescence was detected by exciting the samples at 280nm and ratio of 350nm to 330nm was acquired. Data was fitted to a two-state model. As can be noted in FIG. 4A, chemical stability of Trastuzumab and Trastuzumab-ELP where the ELPs are V(40) (SEQ ID NO: 2), V4A1(40) (SEQ ID NO: 5) and V1A1(40) (SEQ ID NO: 4) fused to the antibody’s HC is very similar. In FIG. 4B we can see the change in Gibbs free energy (AG°) in kcal / mol where the fusion of Trastuzumab’s HC to V1A1(40) (SEQ ID NO: 48) increased the AG° in ~2.5 kcal / mol further stabilizing the protein and the fusion of to V4A1(40) (SEQ ID NO: 45) to Trastuzumab’s HC decreased the AG° in ~1.5 kcal / mol. These results which we believe are driven by the different levels of the ELP’s hydrophobicity, can be correlated with expression, as V1A1(40) (SEQ ID NO: 4) fusions presented higher expression yields while V4A1(40) (SEQ ID NO: 5) presented lower yields.Docket No. 028193-0052-WO01[000135] After we determined that a broad set of ELPs could be expressed and secreted from Expi293 cells, that are cultured at 37°C, we evaluated the use of a different mammalian cell system, which is cultured at 32°C. To this end, we transiently expressed Trastuzumab-ELP HC V(40) (SEQ ID NO: 44) in ExpiCHO. Interestingly, we achieved over 2-fold higher protein yields than when using Expi293 cells. We hypothesize that these higher yields were due to the lower culturing temperature, which reduces the probability of Trastuzumab-ELP transitioning inside the cellular organelles which can block the mammalian cell secretory pathway. FIG. 5A depicts the SDS-Page gel of the Trastuzumab-ELP HC V(40) (SEQ ID NO: 44) following Protein A and size exclusion chromatography (SEC) purification. FIG. 5B depicts the transition temperature characterization for Trastuzumab-ELP HC V(40) (SEQ ID NO: 44). As we can see on the heating curves, the fusion protein can transition at between 35°C - 40°C depending on concentration, which can be one of the factors at play on why culturing cells at lower temperature increases the expression and secretion yield for some of the ELP fusions.[000136] Following the successful expression of 40-mer ELPs fused to the HC and / or LC of Trastuzumab in Expi293 and ExpiCHO cell systems, we sought to evaluate if longer ELPs could also be successfully expressed and secreted. To this end, we transiently transfected DNA coding for ELP HC V(40) and LC into ExpiCHO cells. As can be seen in FIG. 6A, we were able to successfully express and secrete this protein fusion. FIG. 6B depicts the transition temperature characterization for Trastuzumab-ELP HC V4A1(80) (SEQ ID NO: 62). As we can see on the heating curves, the fusion protein can transition at between 35°C - 37°C depending on concentration, which is below the 32°C cell culture incubation temperature.[000137] Following these studies with Trastuzumab fusions we evaluated if other antibodies could also be successfully fused to ELPs. First, we generated a DNA plasmid containing a mouse variable light (VL) and mouse constant kappa light (CL) chains of an in-house developed murine monoclonal antibody (Mouse Antibody-1) terminally fused to an ELP formed by 4 alternating blocks of 20 VPGXG repeats where the guest residues “X” were serine (S) or isoleucine (I). This formed the LC-SISI80 (SEQ ID NO: 102) as shown in subpanel i. of FIG. 7A.This plasmid was transformed into Expi293 cells paired with the original mAb heavy chain (HC) forming a Mouse Antibody(HC+LC-SISI80) as depicted in subpanel ii. of FIG. 7A. Following a round of protein A based purification, we were able to isolate the Mouse Antibody(HC+LC-SISI80) from the culture supernatant as can be seen in the SDS-PAGE gel in subpanel iii. of FIG. 7A, where the purified Mouse Antibody(HC+LC-SISI80) is the band indicated by the light gray, medium gray, and black arrows under non-reducing and reducing conditions. In parallelDocket No. 028193-0052-WO01with this experiment, we also generated a DNA plasmid containing the mouse variable heavy (VH), mouse constant heavy 1 (CH1) a mouse lgG2a Fc chain of the same mouse mAb (Mouse Antibody) fused to the same SISI 80-mer ELP at the C terminus, as shown in subpanel i. of FIG.7B (HC-SISI80 (SEQ ID NO: 100)). This plasmid was co-transformed into Expi293 cells paired with a pcDNA5 plasmid encoding the Mouse Antibody mAb light chain to express Mouse Antibody(HC-SISI80+LC), as depicted in subpanel ii. of FIG. 7B. Following a round of protein A based affinity chromatography, we were able to isolate the Mouse Antibody(HC-SISI80+LC) from the culture supernatant, as shown by the SDS-PAGE gel in subpanel iii. of FIG. 7B, where the purified Mouse Antibody(HC-SISI80+LC) is the band indicated by the light gray, medium gray, and black arrows under non-reducing and reducing conditions.[000138] Following the successful expression and secretion of ELPs fused to the C-terminus of a murine mAb HC and LC, we sought to express ELPs fused to both the heavy and light chains of a mAb. To do so, we transformed Expi293 cells with plasmids containing the SISI 80 ELP at the C-terminus of both the HC and LC, as depicted in FIG. 8A. We also explored in parallel if a third fusion partner could be co-expressed with the mAb-ELP, as this could enable the fusion of other effector molecules to mAbs and potentially the generation of a new class of bi-specific biomolecules that target different molecules simultaneously. As proof-of-principle, we fused mCherry, a hydrophilic and highly stable fluorescent and soluble protein that expresses well in mammalian cells, to the C-terminus of the HC-SISI80 (SEQ ID NO: 100) as depicted in FIG. 8B. This plasmid containing the DNA coding Mouse Antibody mAb HC-SISI80-mCherry fusion was co-transformed into Expi293 cells with a PCDNA5 plasmid encoding the original Mouse Antibody mAb LC, to express Mouse Antibody(HC-S80-mCherry+LC) (SEQ ID NO: 106).[000139] Following protein A affinity and size exclusion (SEC) chromatography of the clarified supernatant of Expi293 cells expressing Mouse Antibody(HC-SISI80+LC-SISI80) (SEQ ID NOs: 100 &102) and Mouse Antibody(HC-S80-mCherry+LC) (SEQ ID NO: 106), we were able to isolate both constructs, as shown in the SDS-PAGE gel (FIG. 8C). To facilitate comparison, we ran in the same gel, the protein A and SEC-purified Mouse Antibody(HC-SISI80+LC) (SEQ ID NO: 100) under non-reducing and reducing conditions in lanes 2 and 3 respectively. Lanes 4 and 5 show the Mouse Antibody(HC-SISI80+LC-SISI80) (SEQ ID NOs: 100 & 102) under nonreducing and reducing conditions and lanes 6 and 7 show Mouse Antibody(HC-S80-mCherry+LC) (SEQ ID NO: 106), also under non-reducing and reducing conditions.Docket No. 028193-0052-WO01[000140] These results demonstrate that ELPs can also be expressed as fusions with murine Abs, and other non-antibody scaffold proteins like mCherry and secreted into the culture supernatant by mammalian cells.[000141] After this demonstration that murine Abs could be expressed in mammalian cells fused to ELPs in different architectures, we investigated if ELPs could also be expressed fused to FcfPs. To this end, we generated DNA plasmids containing the coding sequence for Dulaglutide, a GLP-1 receptor agonist (GLP-1a) fused to a mutated human lgG4 Fc (mhlgG4) that was fused to a wide range of ELPs that were designed for expression in mammalian cells. We expressed Dulaglutide as a control (FIG. 9A-i) and fused to a wide range of ELPs of varying size (from 40 to 120 repeats of the VPGXP repeat) and structure, where the “S” and “I” guest residue 20-repeat blocks were arranged in different configurations to examine the influence of ELP amino acid composition and length on the secretion, phase behavior, and binding of the FcfP-ELP fusions to FcRn and the FcfP target (FIG. 9A-ii to vii). FIG. 9B shows the Dulaglutide control run on an SDS-PAGE gel under non-reducing and reducing conditions in lanes 2 and 3. FIG. 9C depicts the Dulaglutide fused to SI40 (SEQ ID NO: 79), ISIS80 (SEQ ID NO: 80), SSI ISO (SEQ ID NO: 81), I ISS80 (SEQ ID NO: 82) run on an SDS-PAGE gel under non-reducing and reducing conditions and FIG. 9D shows the Dulaglutide fused to SISI80 (SEQ ID NO: 84) and SISIS1120 (SEQ ID NO: 83) also under reducing and non-reducing conditions in SDS-PAGE. These results demonstrate the current capabilities of expressing — and secreting — different ELPs fused to FcfPs, which can have a wide range of applications in biotechnology and the development of therapeutics and drug delivery systems.[000142] Next, we sought to evaluate the expression and secretion of Dulaglutide fused to ELPs of various lengths where valine is the guest residues. As previously described, we transiently transfected DNA coding for these ELP fusions and Dulaglutide in Expi293 cells and Protein A purified the clarified culture supernatant after 7 days of incubation at 37°C. FIG. 10 depicts the SDS-PAGE gel of Dulaglutide control (Trulicity) and the various ELP fusions which were successfully expressed, secreted and purified. Production yields for all constructs varied between 50mg / L to 10Omg / L.[000143] Once we were able to express a well know and FDA approved FcfP biological fused to a wide range of ELPs, we next sought to evaluate if Trastuzumab could also be expressed as SI fusions. Employing published Trastuzumab sequences, we generated a plasmid that encodes Trastuzumab’s HC and LC (FIG. 11 A) and a second plasmid encoding the IISS80 ELPDocket No. 028193-0052-WO01fused to the mAb LC (FIG. 11B). Both sets of plasmids — Trastuzumab (HC+LC) and (HC+LC-IISS80) — were transformed into Expi293 cells to express Trastuzumab and ELP fusions of Trastuzumab, as depicted in subpanel ii. of FIGS. 11A-B. Subpanel iii. of FIGS. 11A-B show the SDS-PAGE gels of the steps related to protein A purification where we highlight the purified Trastuzumab and Trastuzumab-IISS80 (SEQ ID NO: 71), as indicated by the gray arrows.[000144] Next, we generated a plasmid encoding for Trastuzumab HC fused to the IISS80 ELP as shown in FIG. 12A. The plasmids encoding Trastuzumab (HC-IISS80+LC (SEQ ID NO: 70)) were transformed into Expi293 cells forming another architecture of an Ab-ELP fusion, as depicted in FIG. 12B. FIG. 12C shows the SDS-Page gel of the purified Trastuzumab and Trastuzumab (HC-IISS80 (SEQ ID NO: 70)) under reducing and non-reducing conditions.[000145] The results presented in this section demonstrate that we can express and secrete a wide range of ELPs with different architectures and sizes fused to mAbs and FcfPs in mammalian cells. We have also demonstrated proof-of-principle that other proteins can be successfully expressed fused to mAbs-ELPs with widespread potential applications in biotechnology, drug delivery and development of therapeutic biologies.Example 4Description of mAb-ELP and FcfP-ELP phase behavior[000146] From this initial set of expressed mAb- and FcfP-ELP fusions we selected a smaller subset of Dulaglutide-ELP constructs to evaluate if the lower critical solution temperature (LCST) phase transition behavior was preserved. This behavior, which is inherent to ELPs and present in many ELP fusions, can be leveraged: (1) to develop depot forming versions of the FcfP and mAbs with sustained release and overall improved pharmacokinetics; and (2) to purify proteins by inverse transition cycling (ITC), eliminating the need for protein A chromatography. To investigate LCST phase behavior, we measured the optical density at 350 nm as a function of temperature on a UV-vis spectrophotometer for different concentrations of Dulaglutide and various Dulaglutide-ELP fusion (FIGS. 13A-D and FIGS. 14A-D). As can be seen in FIG. 13A, no concentration-dependent change in optical turbidity is observed as a function of concentration for the Dulaglutide control at all concentrations. FIGS. 13B-D show a sharp change in optical turbidity as a function of concentration for Dulaglutide fused to SI40 (SEQ ID NO: 79), ISIS80 (SEQ ID NO: 80) and SSII80 (SEQ ID NO: 81) ELPs, which is indicative of LCST phase transition behavior for these ELP fusions. The temperature at the inflection point inDocket No. 028193-0052-WO01the optical density as a function of temperature is defined as the transition temperature (Tt) — also called the cloud point temperature. Tt shows an inverse dependence on the concentration. Both these features indicate that these ELP fusions undergo phase transition from soluble unimer to a coacervate phase. In contrast, Dulaglutide-IISS80 (SEQ ID NO: 82) shows a very small change in its optical density with temperature, and the temperature at the inflection point of the optical density is largely independent of concentration, indicating that this ELP fusion undergoes a phase transition from soluble unimer to a nanoparticle (FIG. 13E). Similar findings were also seen with the various valine based Dulaglutide fusions (FIGS. 14A-D), where a sharp change in optical turbidity as a function of concentration is dependent on ELP architecture and concentration.[000147] The results presented in this section demonstrated that ELP fusions expressed in mammalian cells may retain phase transition behavior or self-assemble into nanoparticles according to the amino acid composition of the ELP fusion partner.Example 5Description of mAb-ELP and FcfP-ELP and phase behavior-based purification[000148] ELPs are synthetic intrinsically disordered proteins that exhibit tunable, reversible lower critical solution phase behavior. Below their cloud-point temperature (Tt) ELPs are soluble, while above the Tt they phase separate into two immiscible phases: a dense, insoluble, ELP-rich phase composed of micron-sized droplets, and a dilute, ELP-poor phase. The phase separation of ELPs can be triggered by heating the solution above the Tt or by isothermally adding NaCI, which can lower the ELPs’ Tt below the temperature of the solution. The insoluble droplets can be separated from the rest of the solution via centrifugation or tangential flow filtration (TFF) and then resuspended in buffer at a temperature below the ELPs’ Tt, allowing the ELP to return to a soluble state. The resolubilized ELP solution is free of most contaminants present in the original milieu, and cycles of phase transition, separation, and redissolution can be used to purify ELP fusions and their bound partner from complex solutions at large scale without any chromatography steps. We have named this purification process Inverse Transition Cycling (ITC).[000149] In our ITC purification methodology (FIGS. 15A-D), engineered mammalian cells will secrete a mAb or FcfP fused to ELPs into the cell culture supernatant. Then, the cells will be removed via centrifugation or filtration. The LCST phase separation of the mAb-ELP or FcfP-Docket No. 028193-0052-WO01ELP fusion will be triggered by adding 1-2M NaCI, a salt concentration that does not aggregate — “salt-out” — other proteins. The phase-separated droplets will then be separated from the cell culture supernatant via centrifugation or TFF. Next, the complex will be resolubilized in a low salt buffer, typically PBS, and insoluble contaminants will be removed through centrifugation or TFF. Notably, the volume in which the mAb-ELP or FcfP-ELP complex can be resolubilized is largely independent of the original culture volume and is only constrained by the maximum solubility of the complex. Because of this concentration step, all reagents and equipment for subsequent steps can be kept constant. Successive cycles of phase separation followed by solubilization to isolate the mAb-ELP or FcfP-ELP to purity. ITC is easily scalable from milligrams of ELP fusion up to kilograms or greater.[000150] The results presented in this section demonstrate that mAb-ELPs and FcfP-ELPs expressed in mammalian cells can be purified by ITC from culture supernatant.Example 6Description of mAb-ELP / FcfP-ELP binding to target antigen and FcRn[000151] After we demonstrated we could express and purify ELPs fused to mAbs and FcfPs from the culture supernatant with ITC, we used surface plasmon resonance (SPR) to determine the impact of ELPs on mAb target antigen binding. First, we covalently immobilized sGP, the target for Mouse Antibody, in the second channel of a CM5 Biacore T200 chip to a surface density of -500 RU. Next, we injected different concentrations of the wild-type (WT) Mouse Antibody mAb, and its ELP fusions — Mouse Antibody(HC+LC-SISI80) (SEQ ID NO: 102) and Mouse Antibody(HC-SISI80+LC) (SEQ ID NO: 100) for 180 s at 30 pL / min, followed by a 600 s injection of running buffer to measure the binding kinetics of each mAb variant to sGP. FIG. 16A shows the sensorgram and binding affinity (KD) of the original Mouse Antibody mAb and FIGS. 16B-C show the sensorgram and KD of the Mouse Antibody mAb when the SI SI 80 ELP is fused to the LC (SEQ ID NO: 102) or HC (SEQ ID NO: 100) respectively. As can be seen, when fused to the LC, the ELP shows some interference of mAb-Ag binding, which is reflected by a ~6-fold loss in affinity. When fused to the mAb HC, the ELP does not seem to impact the mAb-Ag binding, as the KD of Mouse Antibody and Mouse Antibody(HC-SISI80+LC) (SEQ ID NO: 100) are virtually identical.[000152] Following these initial binding studies with a murine Ab, we investigated the impact of ELP fusion on FcRn mediated mAb recycling by measuring the binding of different mAb-ELPDocket No. 028193-0052-WO01fusions on FcRn binding by SPR. SPR binding studies were performed with biotinylated FcRn that was captured on streptavidin coated CM5 chips with a surface density of ~300-600 RU. Next, we injected different concentrations of two controls, a WT human lgG4 Fc (WT hlgG4) and Dulaglutide (FIGS. 17A-D), which uses a human lgG4 Fc with a series of point mutations to erase its effector function and increase its half-life. WT lgG4 and Dulaglutide were injected for 180 s at 30 pL / min, followed by a 600-s injection of running buffer to establish each mAb’s binding kinetics. Due to the dimeric nature of FcRn-Fc binding, where two FcRns can bind to one Fc, we first fit the acquired sensorgrams with a heterogeneous ligand model (HLM) as can be seen in FIG. 17A and FIG. 17C. Although HLMs are instructive with regard to the multimeric components that form the final interaction, we also fit the data with a steady state model (SSM), as these are recommended to study FcRn binding as they disregard the complexity of the off-rate kinetics and provide an easier way to evaluate FcRn-Fc interactions. FIG. 17B and FIG. 17D show the WT hlgG4 and Dulaglutide sensorgram data fit with a SSM and their calculated KD’s of 114 and 140 nM respectively, which agree with values reported in the literature.[000153] After we established the assay parameters and data-analysis workflow for the controls (WT hlgG4 and Dulaglutide) we performed FcRn binding studies with Dulaglutide SI40 (SEQ ID NO: 79), Dulaglutide IISS80 (SEQ ID NO: 82) and Dulaglutide ISIS80 (SEQ ID NO: 80). FIGS. 18A-F show the SPR sensorgrams for each construct and their respective HLM and SSM fits. Dulaglutide-SI40 (SEQ ID NO: 79) has a slight improvement in its FcRn KD when compared to just Dulaglutide, while Dulaglutide-IISS80 (SEQ ID NO: 82) has a significant decrease (~2-fold) in binding affinity. Interestingly another ELP with the same size but different block architecture (ISIS80 (SEQ ID NO: 80)) has a KD of 89 nM compared to the KD of 140 nM for Dulaglutide, which we speculate is related to the ELP’s improved localized charge profile that translates into a faster / better “on-rate” (FIG. 18E) when compared to the Dulaglutide and Dulaglutide-IISS80 (SEQ ID NO: 82) (FIG. 17C and FIG. 18C).[000154] Finally, to better understand the ELP’s effect on FcRn binding, we also evaluated Dulaglutide SISI80 (SEQ ID NO: 84) and Dulaglutide SISIS1120 (SEQ ID NO: 83) with SPR under the same assay conditions. FIGS. 19A-D show the SPR sensorgrams for each construct and their respective HLM and SSM fits. As can be seen, Dulaglutide-SISI80 (SEQ ID NO: 84) has a slight increase in KD when compared to Dulaglutide-ISIS80 (SEQ ID NO: 80), which indicates a dependence of binding affinity on the ELP architecture, which is further corroborated when analyzing Dulaglutide-SISIS1120 (SEQ ID NO: 83) sensorgrams, which has a bindingDocket No. 028193-0052-WO01affinity of 361 nM that is ~ 3-fold higher than Dulaglutide and Dulaglutide-SISI80 (SEQ ID NO: 84).[000155] The results presented in this section demonstrated that ELPs different sizes, architectures and fusion site can impact binding affinity to antigen target and cell receptors such as FcRn.Example 7Potency of Dulaglutide ELP fusion[000156] Following SPR characterization of the various ELP fusions, we sought to characterize the impact of ELP fusions to Dulaglutide’s half-maximal effective concentration (EC50). To this end, we performed assays were with Chemiluminescent ELISA and GLP-1 receptors were immobilized on MaxiSorp plates. FIG. 20, depicts the EC50 of Trulicity and the different Dulaglutide-ELP fusions assayed. As can be noted, there is an expected increase in EC50 which correlates with the length of the ELP and can be attributed to the steric hinderance imparted by the ELP.Example 8Example Bi-Specific Fusion Proteins[000157] Fusions of ELPs to the C-terminus of either the p35-Fc chain, the p40-Fc chain, or both were produced. Briefly, Expi293 cells were co-transfected with equivalent amounts of plasmid DNA encoding the two chains. Proteins were harvested at 5-7 days post-transfection and purified by Protein A affinity chromatography followed by polishing steps including ion exchange and / or size exclusion as necessary.[000158] FIGS. 21 A-C show SDS PAGE gels of final purified fusions of a representative ELP, V120, to the p35-Fc chain, the p40-Fc chain, or both in the form of heterodimeric IL-12-Fc-ELP fusions under reducing (R) and non-reducing (NR) conditions. Schematics show the design of IL-12-Fc-ELP fusions.[000159] TABLE 1 shows a summary of the results for each of the fusion proteins tested herein.Docket No. 028193-0052-WO01TABLE 1. Summary of the results for each fusion proteinBiologic Heavy Chain Light chain Expressed SEQ ID NO:Trastuzumab V(40) Unmodified YES 42 Trastuzumab Unmodified V(40) YES 43 Trastuzumab V(40) V(40) NO 44 Trastuzumab V4A1(40) Unmodified YES 45 Trastuzumab Unmodified V4A1(40) NO 46 Trastuzumab V4A1(40) V4A1(40) YES 47 Trastuzumab V1A1(40) Unmodified YES 48 Trastuzumab Unmodified V1A1(40) YES 49 Trastuzumab V1A1(40) V1A1(40) YES 50 Trastuzumab V1A4(40) Unmodified YES 51 Trastuzumab Unmodified V1A4(40) YES 52 Trastuzumab V1A4(40) V1A4(40) YES 53 Trastuzumab A(40) Unmodified YES 54 Trastuzumab Unmodified A(40) YES 55 Trastuzumab A(40) A(40) YES 56 Trastuzumab SI(40) Unmodified YES 57 Trastuzumab Unmodified SI(40) YES 58 Trastuzumab SI(40) SI(40) YES 59 Trastuzumab V(60) Unmodified YES 60 Trastuzumab V1A1(60) Unmodified NO 61 Trastuzumab V4A1(80) Unmodified YES 62 Trastuzumab V(80) Unmodified YES 63 Trastuzumab SISI(80) Unmodified YES 64 Trastuzumab Unmodified SISI(80) YES 65 Trastuzumab SISI(80) SISI(80) YES 66 Trastuzumab ISIS(80) Unmodified YES 67 Trastuzumab Unmodified ISIS(80) YES 68 Trastuzumab ISIS(80) ISIS(80) YES 69 Trastuzumab IISS(80) Unmodified YES 70 Trastuzumab Unmodified IISS(80) YES 71 Trastuzumab IISS(80) IISS(80) YES 72 Trastuzumab SSII(80) Unmodified YES 73 Trastuzumab Unmodified SSII(80) YES 74 Trastuzumab SSII(80) SSII(80) YES 75 Trastuzumab SISISI(120) Unmodified NO 76 Trastuzumab Unmodified SISISI(120) NO 77Trastuzumab SISISI(120) SISISI(120) NO 78Docket No. 028193-0052-WO01Dulaglutide SI (40) - YES 79 Dulaglutide ISIS(80) - YES 80 Dulaglutide SSII(80) - YES 81 Dulaglutide IISS(80) - YES 82 Dulaglutide SISISI(120) - YES 83 Dulaglutide SISI(80) - YES 84 Dulaglutide SI-alternating-80 - YES 85 Dulaglutide V40 - YES 86 Dulaglutide V80 - YES 87 Dulaglutide V120 - YES 88 Dulaglutide A1V1(80) - YES 89 Dulaglutide A1V4(80) - YES 90 Dulaglutide S(80) - YES 91 Dulaglutide S(120) - YES 92 Dulaglutide A(80) - YES 93 Dulaglutide A(120) - YES 94 Mouse Antibody S80-mCherry Unmodified YES 106 Mouse Antibody A80-mCherry Unmodified YES 105 Mouse Antibody IVVI80 Unmodified NO 104 Mouse Antibody SISI80 Unmodified YES 100 Mouse Antibody Unmodified S80-mCherry YES 98 Mouse Antibody Unmodified A80-mCherry YES 108 Mouse Antibody Unmodified IVVI80 NO 95 Mouse Antibody S80-mCherry S80-mCherry YES 98 & 106Mouse Antibody SISI80 SISI80 YES 100 & 102[000160] The foregoing description of the specific aspects will so fully reveal the general nature of the disclosed technology that others can, by applying knowledge within the skill of the art, readily modify and / or adapt for various applications such specific aspects, without undue experimentation, without departing from the general concept of the present disclosure.Therefore, such adaptations and modifications are intended to be within the meaning and range of equivalents of the disclosed aspects, based on the teaching and guidance presented herein. It is to be understood that the phraseology or terminology herein is for the purpose of description and not of limitation, such that the terminology or phraseology of the present specification is to be interpreted by the skilled artisan in light of the teachings and guidance.Docket No. 028193-0052-WO01[000161] The breadth and scope of the present disclosure should not be limited by any of the above-described exemplary aspects, but should be defined only in accordance with the following claims and their equivalents.[000162] All publications, patents, patent applications, and / or other documents cited in this application are incorporated by reference in their entirety for all purposes to the same extent as if each individual publication, patent, patent application, and / or other document were individually indicated to be incorporated by reference for all purposes.[000163] For reasons of completeness, the following Embodiments are set out in the following numbered clauses:[000164] Clause 1. A fusion protein comprising: a biologic; and an elastin-like polypeptide (ELP) attached to the biologic, wherein the ELP comprises an amino acid sequence of [VPGXG]m(SEQ ID NO: 1), wherein the ELP has a hydropathicity score of from about 0.5 to about 1.5 and m is an integer from 10 to 120, wherein the fusion protein is expressed and secreted from a eukaryotic cell.[000165] Clause 2. The fusion protein of clause 1, wherein the ELP is attached to the C-terminus of the biologic.[000166] Clause 3. The fusion protein of clause 1 or clause 2, wherein one ELP is attached to the biologic.[000167] Clause 4. The fusion protein of any one of clauses 1-3, wherein more than one ELP is attached to the biologic.[000168] Clause 5. The fusion protein of any one of clauses 1-4, wherein the biologic is an antibody or a Fc-fusion protein.[000169] Clause 6. The fusion protein of clause 5, wherein the antibody is a monoclonal antibody, an immunoglobulin, or a bispecific antibody.[000170] Clause 7. The fusion protein of clause 5 or clause 6, wherein the ELP is attached to a terminus of a heavy chain of the antibody or a terminus of a light chain of the antibody.[000171] Clause 8. The fusion protein of any one of clauses 5-7, wherein the antibody comprises more than one ELP, each ELP being independently attached to a terminus of a heavy chain of the antibody, a terminus of a light chain of the antibody, or a combination thereof.Docket No. 028193-0052-WO01[000172] Clause 9. The fusion protein of clause 8, wherein: the antibody comprises two ELPs, a first ELP attached to a terminus of a first heavy chain and a second ELP attached to a terminus of a second heavy chain; the antibody comprises two ELPs, a first ELP attached to a terminus of a first light chain and a second ELP attached to a terminus of a second light chain; or the antibody comprises four ELPs, a first ELP attached to a terminus of a first heavy chain, a second ELP attached to a terminus of a second heavy chain, a third ELP attached to a terminus of a first light chain, and a fourth ELP attached to a terminus of a second light chain.[000173] Clause 10. The fusion protein of any one of clauses 5-9, wherein: the Fc-fusion protein comprises one ELP attached to a terminus of a first heavy chain or a second heavy chain; or the Fc-fusion protein comprises two ELPs, a first ELP attached to a terminus of a first heavy chain and a second ELP attached to a terminus of a second heavy chain.[000174] Clause 11. The fusion protein of any one of clauses 5-10, wherein the Fc-fusion protein comprises a modified human IgG Fc domain.[000175] Clause 12. The fusion protein of clause 11, wherein the Fc-fusion protein comprises a p35 domain, a p40 domain, an interleukin-12 (IL-12) domain, or a combination thereof.[000176] Clause 13. The fusion protein of any one of clauses 1-12, wherein the ELP comprises an amino acid sequence of [VPGXG]m(SEQ ID NO: 1); wherein X is alanine (Ala), valine (Val), Serine (Ser), Isoleucine (lie), or a mixture thereof; and wherein m is an integer from 20 to 120.[000177] Clause 14. The fusion protein of clause 13, wherein X is a mixture of Ala and Val in a ratio from 10:1 to 1:10 (Ala: Val).[000178] Clause 15. The fusion protein of clause 13 or clause 14, wherein X is a mixture of Ala and Val in a ratio from 4:1 to 1:4 (Ala: Val).[000179] Clause 16. The fusion protein of any one of clauses 13-15, wherein: m is 40 and X is Val or a mixture of Ala and Val in a ratio from 1: 1 to 1:4 (Ala: Val); or m is 80 and X is Ala or a mixture of Ala and Val in a ratio from 1:1 to 4:1 (Ala: Val).[000180] Clause 17. The fusion protein of any one of clauses 13-16, wherein X comprises Ser and lie arranged in blocks, each block comprising about 1 to about 40 repeats of a single residue, and wherein the overall ratio of Ser to Ile residues is from about 10:1 to about 1:10.[000181] Clause 18. The fusion protein of any one of clauses 1-17, wherein the ELP comprises an amino acid sequence of [VPGVG]4o (SEQ ID NO: 2); [VPGAG]4o (SEQ ID NO: 3);[VPG[A1:V1]G]40(SEQ ID NO: 4); [VPG[A1:V4]G]40(SEQ ID NO: 5); [VPG[A4:V1]G]40(SEQ ID NO: 6); [VPGAG]8o (SEQ ID NO: 11); [VPGVG]80(SEQ ID NO: 12); [VPG[A1:V1]G]80(SEQ ID NO: 14);Docket No. 028193-0052-WO01[VPG[A4:V1]G]80(SEQ ID NO: 15);[VPG[A1:V4]G]80(SEQ ID NO: 16); [VPGVG]120(SEQ ID NO: 21); [VPGSG]20-[VPGIG]20-[VPGSG]20-[VPGIG]20(SEQ ID NO: 17); [VPGSG]20-[VPGIG]20-[VPGSG]20-[VPGIG]20-[VPGIG]20(SEQ ID NO: 24); [VPGSG]80(SEQ ID NO: 13); or [VPGIG]40-[VPGSG]40(SEQ ID NO: 115).[000182] Clause 19. The fusion protein of any one of clauses 1-18, wherein the fusion protein has a glycosylation pattern characteristic of the eukaryotic cell.[000183] Clause 20. The fusion protein of any one of clauses 1-19, wherein the ELP has a hydrophobicity effective to promote expression and secretion from the cell without causing precipitation or aggregation.[000184] Clause 21. The fusion protein of any one of clauses 1-20, wherein the ELP has a transition temperature (Tt) that is at least 2°C higher than a temperature that the at least one eukaryotic cell is cultured at.[000185] Clause 22. The fusion protein of any one of clauses 1-21, wherein the ELP has a transition temperature (Tt) of from about 34°C to about 65°C.[000186] Clause 23. The fusion protein of any one of clauses 1-22, wherein the ELP is capable of undergoing a reversible phase transition.[000187] Clause 24. The fusion protein of any one of clauses 1-23, comprising a secondary protein domain attached to the C-terminus of the ELP, wherein the secondary protein domain comprises mCherry, maltose-binding protein (MBP), thioredoxin, N-utilization substance A (NusA), green fluorescent protein (GFP), glutathione S-transferase (GST), an engineered small ubiquitin-like modifier (SUMO) tag, a modified bacterial dehalogenase tag, human serum albumin (HSA), or a second Fc-fusion protein or domain thereof.[000188] Clause 25. The fusion protein of any one of clauses 1-24, wherein the fusion protein comprises a secretion domain attached to the N-terminus of the fusion protein.[000189] Clause 26. The fusion protein of clause 25, wherein the secretion domain comprises an azurocidin signal peptide (SEQ ID NO: 28), an interleukin-2 (IL-2) signal peptide (SEQ ID NO: 31), an albumin signal peptide (SEQ ID NO: 32), a CD33 signal peptide (SEQ ID NO: 33), a tissue plasminogen activator (tPA) signal peptide (SEQ ID NO: 34), a human trypsinogen-2 signal peptide (SEQ ID NO: 35), a human chymotrypsinogen signal peptide (SEQ ID NO: 36), a maltose-binding protein (MBP) signal peptide (SEQ ID NO: 37), a Fc Tag, a glutathione S-transferase (GST) Tag (SEQ ID NO: 40), a CD5 signal peptide (SEQ ID NO: 38), an immunoglobulin Kappa light chain signal peptide (SEQ ID NO: 39), a trypsinogen signal peptide (SEQ ID NO: 29), or a preprolactin signal peptide (SEQ ID NO: 30).Docket No. 028193-0052-WO01[000190] Clause 27. A method of producing a biologic-based fusion protein, comprising: (a) engineering a eukaryotic cell to express a fusion protein comprising a biologic and an elastin-like polypeptide (ELP) attached to a C-terminus of the biologic, wherein the ELP comprises an amino acid sequence of [VPGXG]m(SEQ ID NO: 1), wherein the ELP has a hydropathicity score of from about 0.5 to about 1.5 and m is an integer from 10 to 120; (b) culturing the eukaryotic cell under conditions permitting secretion of the fusion protein into a culture supernatant; and (c) recovering the fusion protein from the culture supernatant.[000191] Clause 28. The method of clause 27, wherein the eukaryotic cell is a mammalian cell or an insect cell.[000192] Clause 29. The method of clause 28, wherein the mammalian cell is a human cell, a mouse cell, a hamster cell, a rat cell, a goat cell, or a rabbit cell.[000193] Clause 30. The method of any one of clauses 27-29, wherein the fusion protein is secreted without intracellular aggregation.[000194] Clause 31. The method of any one of clauses 27-30, wherein the biologic is an antibody or an Fc-fusion protein.[000195] Clause 32. The method of any one of clauses 27-31, wherein the ELP comprises an amino acid sequence of [VPGXG]m(SEQ ID NO: 1); wherein X is alanine (Ala), valine (Val), Serine (Ser), Isoleucine (lie), or a mixture thereof; and wherein m is an integer from 20 to 120.[000196] Clause 33. The method of clause 32, wherein X is a mixture of Ala and Val in a ratio from 10:1 to 1:10 (Ala: Val).[000197] Clause 34. The method of clause 32 or clause 33, wherein X is a mixture of Ala and Val in a ratio from 4:1 to 1:4 (Ala: Val).[000198] Clause 35. The method of any one of clauses 32-34, wherein: m is 40 and X is Val or a mixture of Ala and Val in a ratio from 1: 1 to 1:4 (Ala: Val); or m is 80 and X is Ala or a mixture of Ala and Val in a ratio from 1:1 to 4:1 (Ala: Val).[000199] Clause 36. The method of any one of clauses 32-35, wherein X comprises Ser and lie arranged in blocks, each block comprising about 1 to about 40 repeats of a single residue, and wherein the overall ratio of Ser to Ile residues is from about 10:1 to about 1:10.[000200] Clause 37. The method of any one of clauses 27-36, wherein the ELP has a hydrophobicity effective to promote expression and secretion from the cell without causing precipitation or aggregation.[000201] Clause 38. The method of any one of clauses 27-37, wherein the ELP has a transition temperature (Tt) that is at least 2°C higher than a temperature that the at least one eukaryotic cell is cultured at.Docket No. 028193-0052-WO01[000202] Clause 39. A method of purifying a fusion protein, comprising: (a) providing a culture supernatant comprising the fusion protein of clause 1; (b) inducing a phase transition of the ELP to form an insoluble phase; (c) isolating the insoluble phase from the supernatant; and (d) resolubilizing the fusion protein.[000203] Clause 40. The method of clause 39, wherein the phase transition is induced by increasing temperature of the supernatant above a transition temperature (Tt) of the ELP.[000204] Clause 41. The method of clause 39 or clause 40, wherein the phase transition is induced by adding from about 0.05M to about 5M salt to the supernatant.[000205] Clause 42. The method of clause 41, wherein the salt is sodium chloride, ammonium sulfate, sodium citrate, sodium sulfate, magnesium sulfate, potassium sulfate, sodium phosphates, lithium chloride, sodium acetate, or sodium carbonate.[000206] Clause 43. The method of any one of clauses 39-42, wherein step (c) comprises centrifugation or tangential flow filtration.[000207] Clause 44. The method of any one of clauses 39-43, wherein step (d) comprises adding a low salt buffer to the fusion protein and removing insoluble contaminants using centrifugation or tangential flow filtration.[000208] Clause 45. The method of clause 44, wherein the low salt buffer is phosphate-buffered saline (PBS), Dulbecco’s Phosphate- Buffered Saline (DPBS), citrate buffer, 4-(2-hydroxyethyl)-1 -piperazineethanesulfonic acid (HEPES) buffer, tris(hydroxymethyl)aminomethane (Tris) buffer, acetate buffer, histidine buffer, or succinate buffer.[000209] Clause 46. The method of any one of clauses 39-45, wherein the method is performed without chromatography.[000210] Clause 47. A method of treating a disease or disorder, comprising administering to a subject a therapeutically effective amount of the fusion protein of any one of clauses 1-26.[000211] Clause 48. The method of clause 47, wherein the fusion protein exhibits extended serum half-life relative to the biologic without the ELP.[000212] Clause 49. The method of clause 47 or clause 48, wherein the fusion protein is administered subcutaneously, intramuscularly, or intratumorally; and forms a depot at the administration site.[000213] Clause 50. The method of any one of clauses 47-49, wherein the disease or disorder is cancer, an autoimmune disease, or a metabolic disorder.[000214] Clause 51. A kit, comprising: (a) a fusion protein according to any one of clauses 1-26; and (b) instructions for administering or purifying the fusion protein.Docket No. 028193-0052-WO01[000215] Clause 52. The kit of clause 51, further comprising a buffer suitable for inducing or reversing a phase transition of the ELP, or suitable for in vivo administration.SEQUENCESSEQ ID NO: 1Generic ELP Amino Acid Sequence[VPGXG]mSEQ ID NO: 2ELP Amino Acid Sequence where X is Val and m is 40[VPGVG]40SEQ ID NO: 3ELP Amino Acid Sequence where X is Ala and m is 40[VPGAG]40SEQ ID NO: 4ELP Amino Acid Sequence where X is Ala and Val at a ratio of 1:1 and m is 40 [VPG[A1:V1]G]40SEQ ID NO: 5ELP Amino Acid Sequence where X is Ala and Val at a ratio of 1:4 and m is 40 [VPG[A1:V4]G]40SEQ ID NO: 6ELP Amino Acid Sequence where X is Ala and Val at a ratio of 4:1 and m is 40 [VPG[A4:V1]G]40SEQ ID NO: 7ELP Amino Acid Sequence where X is Ser and He at a ratio of 1:1 and m is 40 [VPG[S1:I1]G]40SEQ ID NO: 8Docket No. 028193-0052-WO01ELP Amino Acid Sequence where X is Val and lie at a ratio of 1: 1 and m is 40 [VPG[V1:I1]G]40SEQ ID NO: 9ELP Amino Acid Sequence where X is Val and m is 60[VPGVG]60SEQ ID NO: 10ELP Amino Acid Sequence where X is Ala and Val at a ratio of 1:1 and m is 60 [VPG[AI: VI]G]60SEQ ID NO: 11ELP Amino Acid Sequence where X is Ala and m is 80[VPGAG]80SEQ ID NO: 12ELP Amino Acid Sequence where X is Val and m is 80[VPGVG]80SEQ ID NO: 13ELP Amino Acid Sequence where X is Ser and m is 80[VPGSG]80SEQ ID NO: 14ELP Amino Acid Sequence where X is Ala and Val at a ratio of 1:1 and m is 80 [VPG[A1:V1]G]80SEQ ID NO: 15ELP Amino Acid Sequence where X is Ala and Val at a ratio of 4:1 and m is 80 [VPG[A4: VI]G]80SEQ ID NO: 16ELP Amino Acid Sequence where X is Ala and Val at a ratio of 1:4 and m is 80[VPG[A1:V4]G]80Docket No. 028193-0052-WO01SEQ ID NO: 17ELP Amino Acid Sequence where X is Ser and lie at a ratio of 1:1 and m is 80[VPGSG]20-[VPGIG]20-[VPGSG]20-[VPGIG]20SEQ ID NO: 18ELP Amino Acid Sequence where X is Ser and lie at a ratio of 1:1 and m is 80[VPGIG]20-[VPGIG]20-[VPGSG]20-[VPGSG]20SEQ ID NO: 19ELP Amino Acid Sequence where X is Ser and lie at a ratio of 1:1 and m is 80[VPGSG]20-[VPGSG]20-[VPGIG]20-[VPGIG]20SEQ ID NO: 20ELP Amino Acid Sequence where X is lie and Val at a ratio of 1:1 and m is 80[VPGIG]20-[VPGVG]20-[VPGVG]20-[VPGIG]20SEQ ID NO: 21ELP Amino Acid Sequence where X is Val and m is 120[VPGVG]120SEQ ID NO: 22ELP Amino Acid Sequence where X is Ala and m is 120[VPGAG]120SEQ ID NO: 23ELP Amino Acid Sequence where X is Ser and m is 120[VPGSG]120SEQ ID NO: 24ELP Amino Acid Sequence where X is Ser and lie at a ratio of 1:1 and m is 120[VPGSG]20-[VPGIG]20-[VPGSG]20-[VPGIG]20-[VPGSG]20-[VPGIG]20SEQ ID NO: 25Docket No. 028193-0052-WO01GS linker between biologic and ELP[GGGGS]i-5SEQ ID NO: 26GS linker between biologic and ELPGGGGSGGGGSGSEQ ID NO: 27mCherry sequence VSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGTQTAKLKVTKGGPLPFAW DILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGVVTVTQDSSLQDGEFIYK VKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKGEIKQRLKLKDGGHYDAEVKTTYKA KKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERAEGRHSTGGMDELYKGSEQ ID NO: 28AZ signal peptideMTRLTVLALLAGLLASSRASEQ ID NO: 29Trypsinogen signal peptideMNLLLILTFVAAAVASEQ ID NO: 30Preprolactin signal peptideMDSKGSSQKGSEQ ID NO: 31IL-2 signal peptideMYRMQLLSCIALSLALVTNSSEQ ID NO: 32Albumin signal peptideMKWVTFISLLFLFSSAYSDocket No. 028193-0052-WO01SEQ ID NO: 33CD33 signal peptideMPLLLLLLPLLWAGALASEQ ID NO: 34tPA signal peptideMDAMKRGLCCVLLLCGAVFVSPSSEQ ID NO: 35Human trypsinogen-2 signal peptideMNLLLILTFVAAAVASEQ ID NO: 36Human chymotrypsinogen signal peptideMAFLWLLSCWALLGTTFGSEQ ID NO: 37MBP signal peptideMKIKTGARILALSALTTMMFSASALASEQ ID NO: 38CD5 signal peptideMPMGSLQPLATLYLLGMLVASCLGSEQ ID NO: 39Immunoglobulin Kappa light chain signal peptideMETPAQLLFLLLLWLPDSTGSEQ ID NO: 40GST Tag MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKKFELGLEFPNLPYYIDGDVK LTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRYGVSRIAYSKDFETLKVDFLSKLPEML KMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVLYMDPMCLDAFPKLVCFKKRIEAIPQIDKYLK SSKYIAWPLQGWQATFGGGDHPPKSDDocket No. 028193-0052-WO01SEQ ID NO: 41Human lgG1 Fc Tag EPKSCDKTHTCPPCPAPELLGGPSVFIFPPKPKDTLMISRTPEVTCVWDVSQDDPEVQFTWYI NNEQVRTARPPLREQQFNSTIRVVSTLPITHQDWLRGKEFKCKVHNKALPAPIEKTISKARGQP REPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFL YSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGKSEQ ID NO: 42Trastuzumab-HC- V(40) \LC EVQLVESGGGLVQPGGSLRLSCAASGFNIKDTYIHWVRQAPGKGLEWVARIYPTNGYTRYADS VKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCSRWGGDGFYAMDYWGQGTLVTVSSASTK GPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSS VVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPK DTLMISRTPEVTCVWDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQ DWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYP SDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHY TQKSLSLSPGKGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPG VGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVP GVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGV PGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGY*DIQMTQSPSSLSASVGDRVTITCRAS QDVNTAVAWYQQKPGKAPKLLIYSASFLYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQ QHYTTPPTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDN ALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGECG*EVQLVESGGGLVQPGGSLRLSCAASGFNIKDTYIHWVRQAPGKGLEWVARIYPTNGYTRYAD SVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCSRWGGDGFYAMDYWGQGTLVTVSSASTK GPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSS VVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPK DTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQ DWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYP SDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHY TQKSLSLSPGKGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPG VGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVP GVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVDocket No. 028193-0052-WO01PGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGY*DIQMTQSPSSLSASVGDRVTITCRAS QDVNTAVAWYQQKPGKAPKLLIYSASFLYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQ QHYTTPPTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDN ALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGECGSEQ ID NO: 43Trastuzumab-HC\LC- V(40) EVQLVESGGGLVQPGGSLRLSCAASGFNIKDTYIHWVRQAPGKGLEWVARIYPTNGYTRYADS VKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCSRWGGDGFYAMDYWGQGTLVTVSSASTK GPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSS VVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPK DTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQ DWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYP SDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHY TQKSLSLSPGKGY*DIQMTQSPSSLSASVGDRVTITCRASQDVNTAVAWYQQKPGKAPKLLIYS ASFLYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQHYTTPPTFGQGTKVEIKRTVAAPS VFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSS TLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGECGVPGVGVPGVGVPGVGVPGVGVPG VGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVP GVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGV PGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGY*EVQLVESGGGLVQPGGSLRLSCAASGFNIKDTYIHWVRQAPGKGLEWVARIYPTNGYTRYA DSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCSRWGGDGFYAMDYWGQGTLVTVSSAST KGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLS SVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKP KDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLH QDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFY PSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNH YTQKSLSLSPGKGY*DIQMTQSPSSLSASVGDRVTITCRASQDVNTAVAWYQQKPGKAPKLLIY SASFLYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQHYTTPPTFGQGTKVEIKRTVAAP SVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLS STLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGECGVPGVGVPGVGVPGVGVPGVGVP GVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGV PGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGDocket No. 028193-0052-WO01VPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGV GY*SEQ ID NO: 44Trastuzumab-HC- V(40) \LC- V(40) EVQLVESGGGLVQPGGSLRLSCAASGFNIKDTYIHWVRQAPGKGLEWVARIYPTNGYTRYADS VKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCSRWGGDGFYAMDYWGQGTLVTVSSASTK GPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSS VVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPK DTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQ DWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYP SDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHY TQKSLSLSPGKGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPG VGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVP GVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGV PGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGY*DIQMTQSPSSLSASVGDRVTITCRAS QDVNTAVAWYQQKPGKAPKLLIYSASFLYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQ QHYTTPPTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDN ALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGECG VPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGV GVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPG VGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVP GVGVPGVGVPGVGVPGVGVPGVGY*EVQLVESGGGLVQPGGSLRLSCAASGFNIKDTYIHWV RQAPGKGLEWVARIYPTNGYTRYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCSRW GGDGFYAMDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSW NSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCD KTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEV HNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQ VYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKL TVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGKGVPGVGVPGVGVPGVGVPGVGVP GVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGV PGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVG VPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGV GY*DIQMTQSPSSLSASVGDRVTITCRASQDVNTAVAWYQQKPGKAPKLLIYSASFLYSGVPSRDocket No. 028193-0052-WO01FSGSRSGTDFTLTISSLQPEDFATYYCQQHYTTPPTFGQGTKVEIKRTVAAPSVFIFPPSDEQLK SGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEK HKVYACEVTHQGLSSPVTKSFNRGECGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPG VGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVP GVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGV PGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGY*SEQ ID NO: 45Trastuzumab-HC-V4A 1-40\LC EVQLVESGGGLVQPGGSLRLSCAASGFNIKDTYIHWVRQAPGKGLEWVARIYPTNGYTRYADS VKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCSRWGGDGFYAMDYWGQGTLVTVSSASTK GPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSS VVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPK DTLMISRTPEVTCVWDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQ DWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYP SDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHY TQKSLSLSPGKGVPGVGVPGVGVPGVGVPGVGVPGAGVPGVGVPGVGVPGVGVPGVGVPG AGVPGVGVPGVGVPGVGVPGVGVPGAGVPGVGVPGVGVPGVGVPGVGVPGAGVPGVGVP GVGVPGVGVPGVGVPGAGVPGVGVPGVGVPGVGVPGVGVPGAGVPGVGVPGVGVPGVGV PGVGVPGAGVPGVGVPGVGVPGVGVPGVGVPGAGY*DIQMTQSPSSLSASVGDRVTITCRAS QDVNTAVAWYQQKPGKAPKLLIYSASFLYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQ QHYTTPPTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDN ALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGECGY*EVQLVESGGGLVQPGGSLRLSCAASGFNIKDTYIHWVRQAPGKGLEWVARIYPTNGYTRYA DSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCSRWGGDGFYAMDYWGQGTLVTVSSAST KGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLS SVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKP KDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLH QDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFY PSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNH YTQKSLSLSPGKGVPGVGVPGVGVPGVGVPGVGVPGAGVPGVGVPGVGVPGVGVPGVGVP GAGVPGVGVPGVGVPGVGVPGVGVPGAGVPGVGVPGVGVPGVGVPGVGVPGAGVPGVGV PGVGVPGVGVPGVGVPGAGVPGVGVPGVGVPGVGVPGVGVPGAGVPGVGVPGVGVPGVG VPGVGVPGAGVPGVGVPGVGVPGVGVPGVGVPGAGY*DIQMTQSPSSLSASVGDRVTITCRADocket No. 028193-0052-WO01SQDVNTAVAWYQQKPGKAPKLLIYSASFLYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQ QHYTTPPTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASWCLLNNFYPREAKVQWKVDN ALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGECGY*SEQ ID NO: 46Trastuzumab-HC\LC-(V4A 1)40 EVQLVESGGGLVQPGGSLRLSCAASGFNIKDTYIHWVRQAPGKGLEWVARIYPTNGYTRYADS VKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCSRWGGDGFYAMDYWGQGTLVTVSSASTK GPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSS VVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPK DTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQ DWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYP SDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHY TQKSLSLSPGKGY*DIQMTQSPSSLSASVGDRVTITCRASQDVNTAVAWYQQKPGKAPKLLIYS ASFLYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQHYTTPPTFGQGTKVEIKRTVAAPS VFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSS TLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGECGVPGVGVPGVGVPGVGVPGVGVPG AGVPGVGVPGVGVPGVGVPGVGVPGAGVPGVGVPGVGVPGVGVPGVGVPGAGVPGVGVP GVGVPGVGVPGVGVPGAGVPGVGVPGVGVPGVGVPGVGVPGAGVPGVGVPGVGVPGVGV PGVGVPGAGVPGVGVPGVGVPGVGVPGVGVPGAGVPGVGVPGVGVPGVGVPGVGVPGAGY*EVQLVESGGGLVQPGGSLRLSCAASGFNIKDTYIHWVRQAPGKGLEWVARIYPTNGYTRYA DSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCSRWGGDGFYAMDYWGQGTLVTVSSAST KGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLS SVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKP KDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLH QDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFY PSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNH YTQKSLSLSPGKGY*DIQMTQSPSSLSASVGDRVTITCRASQDVNTAVAWYQQKPGKAPKLLIY SASFLYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQHYTTPPTFGQGTKVEIKRTVAAP SVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLS STLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGECGVPGVGVPGVGVPGVGVPGVGVP GAGVPGVGVPGVGVPGVGVPGVGVPGAGVPGVGVPGVGVPGVGVPGVGVPGAGVPGVGV PGVGVPGVGVPGVGVPGAGVPGVGVPGVGVPGVGVPGVGVPGAGVPGVGVPGVGVPGVGDocket No. 028193-0052-WO01VPGVGVPGAGVPGVGVPGVGVPGVGVPGVGVPGAGVPGVGVPGVGVPGVGVPGVGVPGA GY*SEQ ID NO: 47Trastuzumab-HC-V4A 1(40)\LC-V4A 1(40) EVQLVESGGGLVQPGGSLRLSCAASGFNIKDTYIHWVRQAPGKGLEWVARIYPTNGYTRYADS VKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCSRWGGDGFYAMDYWGQGTLVTVSSASTK GPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSS VVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPK DTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQ DWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYP SDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHY TQKSLSLSPGKGVPGVGVPGVGVPGVGVPGVGVPGAGVPGVGVPGVGVPGVGVPGVGVPG AGVPGVGVPGVGVPGVGVPGVGVPGAGVPGVGVPGVGVPGVGVPGVGVPGAGVPGVGVP GVGVPGVGVPGVGVPGAGVPGVGVPGVGVPGVGVPGVGVPGAGVPGVGVPGVGVPGVGV PGVGVPGAGVPGVGVPGVGVPGVGVPGVGVPGAGY*DIQMTQSPSSLSASVGDRVTITCRAS QDVNTAVAWYQQKPGKAPKLLIYSASFLYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQ QHYTTPPTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDN ALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGECG VPGVGVPGVGVPGVGVPGVGVPGAGVPGVGVPGVGVPGVGVPGVGVPGAGVPGVGVPGV GVPGVGVPGVGVPGAGVPGVGVPGVGVPGVGVPGVGVPGAGVPGVGVPGVGVPGVGVPG VGVPGAGVPGVGVPGVGVPGVGVPGVGVPGAGVPGVGVPGVGVPGVGVPGVGVPGAGVP GVGVPGVGVPGVGVPGVGVPGAGY*EVQLVESGGGLVQPGGSLRLSCAASGFNIKDTYIHWV RQAPGKGLEWVARIYPTNGYTRYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCSRW GGDGFYAMDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSW NSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCD KTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEV HNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQ VYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKL TVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGKGVPGVGVPGVGVPGVGVPGVGVP GAGVPGVGVPGVGVPGVGVPGVGVPGAGVPGVGVPGVGVPGVGVPGVGVPGAGVPGVGV PGVGVPGVGVPGVGVPGAGVPGVGVPGVGVPGVGVPGVGVPGAGVPGVGVPGVGVPGVG VPGVGVPGAGVPGVGVPGVGVPGVGVPGVGVPGAGVPGVGVPGVGVPGVGVPGVGVPGA GY*DIQMTQSPSSLSASVGDRVTITCRASQDVNTAVAWYQQKPGKAPKLLIYSASFLYSGVPSRDocket No. 028193-0052-WO01FSGSRSGTDFTLTISSLQPEDFATYYCQQHYTTPPTFGQGTKVEIKRTVAAPSVFIFPPSDEQLK SGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEK HKVYACEVTHQGLSSPVTKSFNRGECGVPGVGVPGVGVPGVGVPGVGVPGAGVPGVGVPG VGVPGVGVPGVGVPGAGVPGVGVPGVGVPGVGVPGVGVPGAGVPGVGVPGVGVPGVGVP GVGVPGAGVPGVGVPGVGVPGVGVPGVGVPGAGVPGVGVPGVGVPGVGVPGVGVPGAGV PGVGVPGVGVPGVGVPGVGVPGAGVPGVGVPGVGVPGVGVPGVGVPGAGY*SEQ ID NO: 48Trastuzumab-HC-V1A 1(40)\LC EVQLVESGGGLVQPGGSLRLSCAASGFNIKDTYIHWVRQAPGKGLEWVARIYPTNGYTRYADS VKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCSRWGGDGFYAMDYWGQGTLVTVSSASTK GPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSS VVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPK DTLMISRTPEVTCVWDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQ DWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYP SDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHY TQKSLSLSPGKGVPGVGVPGAGVPGVGVPGAGVPGVGVPGAGVPGVGVPGAGVPGVGVPG AGVPGVGVPGAGVPGVGVPGAGVPGVGVPGAGVPGVGVPGAGVPGVGVPGAGVPGVGVP GAGVPGVGVPGAGVPGVGVPGAGVPGVGVPGAGVPGVGVPGAGVPGVGVPGAGVPGVGV PGAGVPGVGVPGAGVPGVGVPGAGVPGVGVPGAGY*DIQMTQSPSSLSASVGDRVTITCRAS QDVNTAVAWYQQKPGKAPKLLIYSASFLYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQ QHYTTPPTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDN ALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGECGY*EVQLVESGGGLVQPGGSLRLSCAASGFNIKDTYIHWVRQAPGKGLEWVARIYPTNGYTRYA DSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCSRWGGDGFYAMDYWGQGTLVTVSSAST KGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLS SVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKP KDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLH QDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFY PSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNH YTQKSLSLSPGKGVPGVGVPGAGVPGVGVPGAGVPGVGVPGAGVPGVGVPGAGVPGVGVP GAGVPGVGVPGAGVPGVGVPGAGVPGVGVPGAGVPGVGVPGAGVPGVGVPGAGVPGVGV PGAGVPGVGVPGAGVPGVGVPGAGVPGVGVPGAGVPGVGVPGAGVPGVGVPGAGVPGVG VPGAGVPGVGVPGAGVPGVGVPGAGVPGVGVPGAGY*DIQMTQSPSSLSASVGDRVTITCRADocket No. 028193-0052-WO01SQDVNTAVAWYQQKPGKAPKLLIYSASFLYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQ QHYTTPPTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASWCLLNNFYPREAKVQWKVDN ALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGECGY*SEQ ID NO: 49Trastuzumab-HC|LC-(V1A1)40 EVQLVESGGGLVQPGGSLRLSCAASGFNIKDTYIHWVRQAPGKGLEWVARIYPTNGYTRYADS VKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCSRWGGDGFYAMDYWGQGTLVTVSSASTK GPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSS VVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPK DTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQ DWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYP SDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHY TQKSLSLSPGKGY*DIQMTQSPSSLSASVGDRVTITCRASQDVNTAVAWYQQKPGKAPKLLIYS ASFLYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQHYTTPPTFGQGTKVEIKRTVAAPS VFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSS TLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGECGVPGVGVPGAGVPGVGVPGAGVPG VGVPGAGVPGVGVPGAGVPGVGVPGAGVPGVGVPGAGVPGVGVPGAGVPGVGVPGAGVP GVGVPGAGVPGVGVPGAGVPGVGVPGAGVPGVGVPGAGVPGVGVPGAGVPGVGVPGAGV PGVGVPGAGVPGVGVPGAGVPGVGVPGAGVPGVGVPGAGVPGVGVPGAGVPGVGVPGAGY*EVQLVESGGGLVQPGGSLRLSCAASGFNIKDTYIHWVRQAPGKGLEWVARIYPTNGYTRYA DSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCSRWGGDGFYAMDYWGQGTLVTVSSAST KGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLS SVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKP KDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLH QDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFY PSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNH YTQKSLSLSPGKGY*DIQMTQSPSSLSASVGDRVTITCRASQDVNTAVAWYQQKPGKAPKLLIY SASFLYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQHYTTPPTFGQGTKVEIKRTVAAP SVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLS STLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGECGVPGVGVPGAGVPGVGVPGAGVP GVGVPGAGVPGVGVPGAGVPGVGVPGAGVPGVGVPGAGVPGVGVPGAGVPGVGVPGAGV PGVGVPGAGVPGVGVPGAGVPGVGVPGAGVPGVGVPGAGVPGVGVPGAGVPGVGVPGAGDocket No. 028193-0052-WO01VPGVGVPGAGVPGVGVPGAGVPGVGVPGAGVPGVGVPGAGVPGVGVPGAGVPGVGVPGA GY*SEQ ID NO: 50Trastuzumab-HC-V1A1(40)|LC-V1A1(40) EVQLVESGGGLVQPGGSLRLSCAASGFNIKDTYIHWVRQAPGKGLEWVARIYPTNGYTRYADS VKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCSRWGGDGFYAMDYWGQGTLVTVSSASTK GPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSS VVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPK DTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQ DWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYP SDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHY TQKSLSLSPGKGVPGVGVPGAGVPGVGVPGAGVPGVGVPGAGVPGVGVPGAGVPGVGVPG AGVPGVGVPGAGVPGVGVPGAGVPGVGVPGAGVPGVGVPGAGVPGVGVPGAGVPGVGVP GAGVPGVGVPGAGVPGVGVPGAGVPGVGVPGAGVPGVGVPGAGVPGVGVPGAGVPGVGV PGAGVPGVGVPGAGVPGVGVPGAGVPGVGVPGAGY*DIQMTQSPSSLSASVGDRVTITCRAS QDVNTAVAWYQQKPGKAPKLLIYSASFLYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQ QHYTTPPTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDN ALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGECG VPGVGVPGAGVPGVGVPGAGVPGVGVPGAGVPGVGVPGAGVPGVGVPGAGVPGVGVPGA GVPGVGVPGAGVPGVGVPGAGVPGVGVPGAGVPGVGVPGAGVPGVGVPGAGVPGVGVPG AGVPGVGVPGAGVPGVGVPGAGVPGVGVPGAGVPGVGVPGAGVPGVGVPGAGVPGVGVP GAGVPGVGVPGAGVPGVGVPGAGY*EVQLVESGGGLVQPGGSLRLSCAASGFNIKDTYIHWV RQAPGKGLEWVARIYPTNGYTRYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCSRW GGDGFYAMDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSW NSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCD KTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEV HNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQ VYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKL TVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGKGVPGVGVPGAGVPGVGVPGAGVP GVGVPGAGVPGVGVPGAGVPGVGVPGAGVPGVGVPGAGVPGVGVPGAGVPGVGVPGAGV PGVGVPGAGVPGVGVPGAGVPGVGVPGAGVPGVGVPGAGVPGVGVPGAGVPGVGVPGAG VPGVGVPGAGVPGVGVPGAGVPGVGVPGAGVPGVGVPGAGVPGVGVPGAGVPGVGVPGA GY*DIQMTQSPSSLSASVGDRVTITCRASQDVNTAVAWYQQKPGKAPKLLIYSASFLYSGVPSRDocket No. 028193-0052-WO01FSGSRSGTDFTLTISSLQPEDFATYYCQQHYTTPPTFGQGTKVEIKRTVAAPSVFIFPPSDEQLK SGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEK HKVYACEVTHQGLSSPVTKSFNRGECGVPGVGVPGAGVPGVGVPGAGVPGVGVPGAGVPG VGVPGAGVPGVGVPGAGVPGVGVPGAGVPGVGVPGAGVPGVGVPGAGVPGVGVPGAGVP GVGVPGAGVPGVGVPGAGVPGVGVPGAGVPGVGVPGAGVPGVGVPGAGVPGVGVPGAGV PGVGVPGAGVPGVGVPGAGVPGVGVPGAGVPGVGVPGAGVPGVGVPGAGY*SEQ ID NO: 51Trastuzumab-HC V1A4(40)|LC EVQLVESGGGLVQPGGSLRLSCAASGFNIKDTYIHWVRQAPGKGLEWVARIYPTNGYTRYADS VKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCSRWGGDGFYAMDYWGQGTLVTVSSASTK GPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSS VVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPK DTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGKGVPGVGVPGAGVPGAGVPGAGVPGAGVPGVGVPGAGVPGAGVPGAGVPG AGVPGVGVPGAGVPGAGVPGAGVPGAGVPGVGVPGAGVPGAGVPGAGVPGAGVPGVGVP GAGVPGAGVPGAGVPGAGVPGVGVPGAGVPGAGVPGAGVPGAGVPGVGVPGAGVPGAGV PGAGVPGAGVPGVGVPGAGVPGAGVPGAGVPGAGY*DIQMTQSPSSLSASVGDRVTITCRAS QDVNTAVAWYQQKPGKAPKLLIYSASFLYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQ QHYTTPPTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDN ALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGECGY*EVQLVESGGGLVQPGGSLRLSCAASGFNIKDTYIHWVRQAPGKGLEWVARIYPTNGYTRYA DSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCSRWGGDGFYAMDYWGQGTLVTVSSAST KGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLS SVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKP KDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLH QDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFY PSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNH YTQKSLSLSPGKGVPGVGVPGAGVPGAGVPGAGVPGAGVPGVGVPGAGVPGAGVPGAGVP GAGVPGVGVPGAGVPGAGVPGAGVPGAGVPGVGVPGAGVPGAGVPGAGVPGAGVPGVGV PGAGVPGAGVPGAGVPGAGVPGVGVPGAGVPGAGVPGAGVPGAGVPGVGVPGAGVPGAG VPGAGVPGAGVPGVGVPGAGVPGAGVPGAGVPGAGY*DIQMTQSPSSLSASVGDRVTITCRADocket No. 028193-0052-WO01SQDVNTAVAWYQQKPGKAPKLLIYSASFLYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQ QHYTTPPTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASWCLLNNFYPREAKVQWKVDN ALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGECGY*SEQ ID NO: 52Trastuzumab-HC|LC-V1A4(40) EVQLVESGGGLVQPGGSLRLSCAASGFNIKDTYIHWVRQAPGKGLEWVARIYPTNGYTRYADS VKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCSRWGGDGFYAMDYWGQGTLVTVSSASTK GPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSS VVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPK DTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQ DWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYP SDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHY TQKSLSLSPGKGY*DIQMTQSPSSLSASVGDRVTITCRASQDVNTAVAWYQQKPGKAPKLLIYS ASFLYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQHYTTPPTFGQGTKVEIKRTVAAPS VFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSS TLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGECGVPGVGVPGAGVPGAGVPGAGVPG AGVPGVGVPGAGVPGAGVPGAGVPGAGVPGVGVPGAGVPGAGVPGAGVPGAGVPGVGVP GAGVPGAGVPGAGVPGAGVPGVGVPGAGVPGAGVPGAGVPGAGVPGVGVPGAGVPGAGV PGAGVPGAGVPGVGVPGAGVPGAGVPGAGVPGAGVPGVGVPGAGVPGAGVPGAGVPGAGY*EVQLVESGGGLVQPGGSLRLSCAASGFNIKDTYIHWVRQAPGKGLEWVARIYPTNGYTRYA DSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCSRWGGDGFYAMDYWGQGTLVTVSSAST KGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLS SVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKP KDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLH QDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFY PSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNH YTQKSLSLSPGKGY*DIQMTQSPSSLSASVGDRVTITCRASQDVNTAVAWYQQKPGKAPKLLIY SASFLYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQHYTTPPTFGQGTKVEIKRTVAAP SVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLS STLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGECGVPGVGVPGAGVPGAGVPGAGVP GAGVPGVGVPGAGVPGAGVPGAGVPGAGVPGVGVPGAGVPGAGVPGAGVPGAGVPGVGV PGAGVPGAGVPGAGVPGAGVPGVGVPGAGVPGAGVPGAGVPGAGVPGVGVPGAGVPGAGDocket No. 028193-0052-WO01VPGAGVPGAGVPGVGVPGAGVPGAGVPGAGVPGAGVPGVGVPGAGVPGAGVPGAGVPGA GY*SEQ ID NO: 53Trastuzumab-HC V1A4(40)|LC-V1A4(40) EVQLVESGGGLVQPGGSLRLSCAASGFNIKDTYIHWVRQAPGKGLEWVARIYPTNGYTRYADS VKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCSRWGGDGFYAMDYWGQGTLVTVSSASTK GPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSS VVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPK DTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQ DWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYP SDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHY TQKSLSLSPGKGVPGVGVPGAGVPGAGVPGAGVPGAGVPGVGVPGAGVPGAGVPGAGVPG AGVPGVGVPGAGVPGAGVPGAGVPGAGVPGVGVPGAGVPGAGVPGAGVPGAGVPGVGVP GAGVPGAGVPGAGVPGAGVPGVGVPGAGVPGAGVPGAGVPGAGVPGVGVPGAGVPGAGV PGAGVPGAGVPGVGVPGAGVPGAGVPGAGVPGAGY*DIQMTQSPSSLSASVGDRVTITCRAS QDVNTAVAWYQQKPGKAPKLLIYSASFLYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQ QHYTTPPTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDN ALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGECG VPGVGVPGAGVPGAGVPGAGVPGAGVPGVGVPGAGVPGAGVPGAGVPGAGVPGVGVPGA GVPGAGVPGAGVPGAGVPGVGVPGAGVPGAGVPGAGVPGAGVPGVGVPGAGVPGAGVPG AGVPGAGVPGVGVPGAGVPGAGVPGAGVPGAGVPGVGVPGAGVPGAGVPGAGVPGAGVP GVGVPGAGVPGAGVPGAGVPGAGY*EVQLVESGGGLVQPGGSLRLSCAASGFNIKDTYIHWV RQAPGKGLEWVARIYPTNGYTRYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCSRW GGDGFYAMDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSW NSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCD KTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEV HNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQ VYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKL TVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGKGVPGVGVPGAGVPGAGVPGAGVP GAGVPGVGVPGAGVPGAGVPGAGVPGAGVPGVGVPGAGVPGAGVPGAGVPGAGVPGVGV PGAGVPGAGVPGAGVPGAGVPGVGVPGAGVPGAGVPGAGVPGAGVPGVGVPGAGVPGAG VPGAGVPGAGVPGVGVPGAGVPGAGVPGAGVPGAGVPGVGVPGAGVPGAGVPGAGVPGA GY*DIQMTQSPSSLSASVGDRVTITCRASQDVNTAVAWYQQKPGKAPKLLIYSASFLYSGVPSRDocket No. 028193-0052-WO01FSGSRSGTDFTLTISSLQPEDFATYYCQQHYTTPPTFGQGTKVEIKRTVAAPSVFIFPPSDEQLK SGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEK HKVYACEVTHQGLSSPVTKSFNRGECGVPGVGVPGAGVPGAGVPGAGVPGAGVPGVGVPG AGVPGAGVPGAGVPGAGVPGVGVPGAGVPGAGVPGAGVPGAGVPGVGVPGAGVPGAGVP GAGVPGAGVPGVGVPGAGVPGAGVPGAGVPGAGVPGVGVPGAGVPGAGVPGAGVPGAGV PGVGVPGAGVPGAGVPGAGVPGAGVPGVGVPGAGVPGAGVPGAGVPGAGY*SEQ ID NO: 54Trastuzumab-HC-A(40)|LC EVQLVESGGGLVQPGGSLRLSCAASGFNIKDTYIHWVRQAPGKGLEWVARIYPTNGYTRYADS VKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCSRWGGDGFYAMDYWGQGTLVTVSSASTK GPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSS VVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPK DTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGKGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPG AGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVP GAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGV PGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGY*DIQMTQSPSSLSASVGDRVTITCRAS QDVNTAVAWYQQKPGKAPKLLIYSASFLYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQ QHYTTPPTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDN ALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGECGY*EVQLVESGGGLVQPGGSLRLSCAASGFNIKDTYIHWVRQAPGKGLEWVARIYPTNGYTRYA DSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCSRWGGDGFYAMDYWGQGTLVTVSSAST KGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLS SVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKP KDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLH QDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFY PSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNH YTQKSLSLSPGKGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVP GAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGV PGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAG VPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGY*DIQMTQSPSSLSASVGDRVTITCRADocket No. 028193-0052-WO01SQDVNTAVAWYQQKPGKAPKLLIYSASFLYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQ QHYTTPPTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASWCLLNNFYPREAKVQWKVDN ALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGECGY*SEQ ID NO: 55Trastuzumab-HC|LC-A(40) EVQLVESGGGLVQPGGSLRLSCAASGFNIKDTYIHWVRQAPGKGLEWVARIYPTNGYTRYADS VKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCSRWGGDGFYAMDYWGQGTLVTVSSASTK GPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSS VVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPK DTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQ DWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYP SDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHY TQKSLSLSPGKGY*DIQMTQSPSSLSASVGDRVTITCRASQDVNTAVAWYQQKPGKAPKLLIYS ASFLYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQHYTTPPTFGQGTKVEIKRTVAAPS VFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSS TLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGECGVPGAGVPGAGVPGAGVPGAGVPG AGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVP GAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGV PGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGY*EVQLVESGGGLVQPGGSLRLSCAASGFNIKDTYIHWVRQAPGKGLEWVARIYPTNGYTRYA DSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCSRWGGDGFYAMDYWGQGTLVTVSSAST KGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLS SVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKP KDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLH QDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFY PSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNH YTQKSLSLSPGKGY*DIQMTQSPSSLSASVGDRVTITCRASQDVNTAVAWYQQKPGKAPKLLIY SASFLYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQHYTTPPTFGQGTKVEIKRTVAAP SVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLS STLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGECGVPGAGVPGAGVPGAGVPGAGVP GAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGV PGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGDocket No. 028193-0052-WO01VPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGA GY*SEQ ID NO: 56Trastuzumab-HC-A (40) |LC-A (40) EVQLVESGGGLVQPGGSLRLSCAASGFNIKDTYIHWVRQAPGKGLEWVARIYPTNGYTRYADS VKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCSRWGGDGFYAMDYWGQGTLVTVSSASTK GPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSS VVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPK DTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQ DWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYP SDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHY TQKSLSLSPGKGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPG AGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVP GAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGV PGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGY*DIQMTQSPSSLSASVGDRVTITCRAS QDVNTAVAWYQQKPGKAPKLLIYSASFLYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQ QHYTTPPTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDN ALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGECG VPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGA GVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPG AGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVP GAGVPGAGVPGAGVPGAGVPGAGY*EVQLVESGGGLVQPGGSLRLSCAASGFNIKDTYIHWV RQAPGKGLEWVARIYPTNGYTRYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCSRW GGDGFYAMDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSW NSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCD KTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEV HNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQ VYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKL TVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGKGVPGAGVPGAGVPGAGVPGAGVP GAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGV PGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAG VPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGA GY*DIQMTQSPSSLSASVGDRVTITCRASQDVNTAVAWYQQKPGKAPKLLIYSASFLYSGVPSRDocket No. 028193-0052-WO01FSGSRSGTDFTLTISSLQPEDFATYYCQQHYTTPPTFGQGTKVEIKRTVAAPSVFIFPPSDEQLK SGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEK HKVYACEVTHQGLSSPVTKSFNRGECGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPG AGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVP GAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGV PGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGY*SEQ ID NO: 57Trastuzumab-HC-SI(40)|LC EVQLVESGGGLVQPGGSLRLSCAASGFNIKDTYIHWVRQAPGKGLEWVARIYPTNGYTRYADS VKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCSRWGGDGFYAMDYWGQGTLVTVSSASTK GPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSS VVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPK DTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGKGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPG SGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGIGVPG IGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIG VPGIGVPGIGVPGIGVPGIGVPGIGY*DIQMTQSPSSLSASVGDRVTITCRASQDVNTAVAWYQ QKPGKAPKLLIYSASFLYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQHYTTPPTFGQG TKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTE QDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC*EVQLVESGGGLVQ PGGSLRLSCAASGFNIKDTYIHWVRQAPGKGLEWVARIYPTNGYTRYADSVKGRFTISADTSK NTAYLQMNSLRAEDTAVYYCSRWGGDGFYAMDYWGQGTLVTVSSASTKGPSVFPLAPSSKS TSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQT YICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTC VVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCK VSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQ PENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGKG VPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGS GVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGIGVPGIGVPGIGVPGIG VPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVP GIGVPGIGVPGIGY*DIQMTQSPSSLSASVGDRVTITCRASQDVNTAVAWYQQKPGKAPKLLIYSDocket No. 028193-0052-WO01ASFLYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQHYTTPPTFGQGTKVEIKRTVAAPS VFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSS TLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC*SEQ ID NO: 58Trastuzumab-HC|LC-SI(40) EVQLVESGGGLVQPGGSLRLSCAASGFNIKDTYIHWVRQAPGKGLEWVARIYPTNGYTRYADS VKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCSRWGGDGFYAMDYWGQGTLVTVSSASTK GPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSS VVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPK DTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQ DWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYP SDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHY TQKSLSLSPGKGY*DIQMTQSPSSLSASVGDRVTITCRASQDVNTAVAWYQQKPGKAPKLLIYS ASFLYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQHYTTPPTFGQGTKVEIKRTVAAPS VFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSS TLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGECGVPGSGVPGSGVPGSGVPGSGVPG SGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVP GSGVPGSGVPGSGVPGSGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGV PGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGY*EVQLVESGGG LVQPGGSLRLSCAASGFNIKDTYIHWVRQAPGKGLEWVARIYPTNGYTRYADSVKGRFTISADT SKNTAYLQMNSLRAEDTAVYYCSRWGGDGFYAMDYWGQGTLVTVSSASTKGPSVFPLAPSS KSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGT QTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEV TCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYK CKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESN GQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG KGY*DIQMTQSPSSLSASVGDRVTITCRASQDVNTAVAWYQQKPGKAPKLLIYSASFLYSGVPS RFSGSRSGTDFTLTISSLQPEDFATYYCQQHYTTPPTFGQGTKVEIKRTVAAPSVFIFPPSDEQL KSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEK HKVYACEVTHQGLSSPVTKSFNRGECGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPG SGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVP GSGVPGSGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVP GIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGY*Docket No. 028193-0052-WO01SEQ ID NO: 59Trastuzumab-HC- SI(40)\LC- SI(40) EVQLVESGGGLVQPGGSLRLSCAASGFNIKDTYIHWVRQAPGKGLEWVARIYPTNGYTRYADS VKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCSRWGGDGFYAMDYWGQGTLVTVSSASTK GPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSS VVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPK DTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGKGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPG SGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGIGVPG IGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIG VPGIGVPGIGVPGIGVPGIGVPGIGY*DIQMTQSPSSLSASVGDRVTITCRASQDVNTAVAWYQ QKPGKAPKLLIYSASFLYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQHYTTPPTFGQG TKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTE QDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGECGVPGSGVPGSGVP GSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGV PGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGI GVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGY*EVQLVESGGGLVQPGGSLRLSCAASGFNIKDTYIHWVRQAPGKGLEWVARIYPTNGYTRYA DSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCSRWGGDGFYAMDYWGQGTLVTVSSAST KGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLS SVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKP KDTLMISRTPEVTCWVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLH QDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFY PSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNH YTQKSLSLSPGKGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVP GSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGIGVP GIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGI GVPGIGVPGIGVPGIGVPGIGVPGIGY*DIQMTQSPSSLSASVGDRVTITCRASQDVNTAVAWY QQKPGKAPKLLIYSASFLYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQHYTTPPTFGQ GTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESV TEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGECGVPGSGVPGSGDocket No. 028193-0052-WO01VPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGS GVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVP GIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGI GY*SEQ ID NO: 60Trastuzumab-HC-V(60)\LC EVQLVESGGGLVQPGGSLRLSCAASGFNIKDTYIHWVRQAPGKGLEWVARIYPTNGYTRYADS VKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCSRWGGDGFYAMDYWGQGTLVTVSSASTK GPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSS VVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPK DTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQ DWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYP SDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHY TQKSLSLSPGKGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPG VGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVP GVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGV PGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVG VPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGV GVPGVGVPGVGVPGVGY*DIQMTQSPSSLSASVGDRVTITCRASQDVNTAVAWYQQKPGKAP KLLIYSASFLYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQHYTTPPTFGQGTKVEIKRT VAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDST YSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGECGY*EVQLVESGGGLVQPGGSL RLSCAASGFNIKDTYIHWVRQAPGKGLEWVARIYPTNGYTRYADSVKGRFTISADTSKNTAYLQ MNSLRAEDTAVYYCSRWGGDGFYAMDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTA ALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVN HKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVS HEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALP APIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKT TPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGKGVPGVGVP GVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGV PGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVG VPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGV GVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGDocket No. 028193-0052-WO01VGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGY*D IQMTQSPSSLSASVGDRVTITCRASQDVNTAVAWYQQKPGKAPKLLIYSASFLYSGVPSRFSG SRSGTDFTLTISSLQPEDFATYYCQQHYTTPPTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGT ASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVY ACEVTHQGLSSPVTKSFNRGECGY*SEQ ID NO: 61Trastuzumab-HC-V1A 1(60)\LC EVQLVESGGGLVQPGGSLRLSCAASGFNIKDTYIHWVRQAPGKGLEWVARIYPTNGYTRYADS VKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCSRWGGDGFYAMDYWGQGTLVTVSSASTK GPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSS VVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPK DTLMISRTPEVTCVWDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQ DWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYP SDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHY TQKSLSLSPGKGVPGVGVPGAGVPGVGVPGAGVPGVGVPGAGVPGVGVPGAGVPGVGVPG AGVPGVGVPGAGVPGVGVPGAGVPGVGVPGAGVPGVGVPGAGVPGVGVPGAGVPGVGVP GAGVPGVGVPGAGVPGVGVPGAGVPGVGVPGAGVPGVGVPGAGVPGVGVPGAGVPGVGV PGAGVPGVGVPGAGVPGVGVPGAGVPGVGVPGAGVPGVGVPGAGVPGVGVPGAGVPGVG VPGAGVPGVGVPGAGVPGVGVPGAGVPGVGVPGAGVPGVGVPGAGVPGVGVPGAGVPGV GVPGAGVPGVGVPGAGY*DIQMTQSPSSLSASVGDRVTITCRASQDVNTAVAWYQQKPGKAP KLLIYSASFLYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQHYTTPPTFGQGTKVEIKRT VAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDST YSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGECGY*EVQLVESGGGLVQPGGSL RLSCAASGFNIKDTYIHWVRQAPGKGLEWVARIYPTNGYTRYADSVKGRFTISADTSKNTAYLQ MNSLRAEDTAVYYCSRWGGDGFYAMDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTA ALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVN HKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVS HEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALP APIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKT TPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGKGVPGVGVP GAGVPGVGVPGAGVPGVGVPGAGVPGVGVPGAGVPGVGVPGAGVPGVGVPGAGVPGVGV PGAGVPGVGVPGAGVPGVGVPGAGVPGVGVPGAGVPGVGVPGAGVPGVGVPGAGVPGVG VPGAGVPGVGVPGAGVPGVGVPGAGVPGVGVPGAGVPGVGVPGAGVPGVGVPGAGVPGVDocket No. 028193-0052-WO01GVPGAGVPGVGVPGAGVPGVGVPGAGVPGVGVPGAGVPGVGVPGAGVPGVGVPGAGVPG VGVPGAGVPGVGVPGAGVPGVGVPGAGVPGVGVPGAGVPGVGVPGAGVPGVGVPGAGY*D IQMTQSPSSLSASVGDRVTITCRASQDVNTAVAWYQQKPGKAPKLLIYSASFLYSGVPSRFSG SRSGTDFTLTISSLQPEDFATYYCQQHYTTPPTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGT ASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVY ACEVTHQGLSSPVTKSFNRGECGY*SEQ ID NO: 62Trastuzumab-HC-V4A 1(80)\LC EVQLVESGGGLVQPGGSLRLSCAASGFNIKDTYIHWVRQAPGKGLEWVARIYPTNGYTRYADS VKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCSRWGGDGFYAMDYWGQGTLVTVSSASTK GPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSS VVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPK DTLMISRTPEVTCVWDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQ DWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYP SDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHY TQKSLSLSPGKGVPGVGVPGVGVPGVGVPGVGVPGAGVPGVGVPGVGVPGVGVPGVGVPG AGVPGVGVPGVGVPGVGVPGVGVPGAGVPGVGVPGVGVPGVGVPGVGVPGAGVPGVGVP GVGVPGVGVPGVGVPGAGVPGVGVPGVGVPGVGVPGVGVPGAGVPGVGVPGVGVPGVGV PGVGVPGAGVPGVGVPGVGVPGVGVPGVGVPGAGVPGVGVPGVGVPGVGVPGVGVPGAG VPGVGVPGVGVPGVGVPGVGVPGAGVPGVGVPGVGVPGVGVPGVGVPGAGVPGVGVPGV GVPGVGVPGVGVPGAGVPGVGVPGVGVPGVGVPGVGVPGAGVPGVGVPGVGVPGVGVPG VGVPGAGVPGVGVPGVGVPGVGVPGVGVPGAGVPGVGVPGVGVPGVGVPGVGVPGAGY*D IQMTQSPSSLSASVGDRVTITCRASQDVNTAVAWYQQKPGKAPKLLIYSASFLYSGVPSRFSG SRSGTDFTLTISSLQPEDFATYYCQQHYTTPPTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGT ASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVY ACEVTHQGLSSPVTKSFNRGECGY*EVQLVESGGGLVQPGGSLRLSCAASGFNIKDTYIHWVR QAPGKGLEWVARIYPTNGYTRYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCSRWG GDGFYAMDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWN SGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKT HTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCWVDVSHEDPEVKFNWYVDGVEVHN AKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVY TLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTV DKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGKGVPGVGVPGVGVPGVGVPGVGVPGADocket No. 028193-0052-WO01GVPGVGVPGVGVPGVGVPGVGVPGAGVPGVGVPGVGVPGVGVPGVGVPGAGVPGVGVPG VGVPGVGVPGVGVPGAGVPGVGVPGVGVPGVGVPGVGVPGAGVPGVGVPGVGVPGVGVP GVGVPGAGVPGVGVPGVGVPGVGVPGVGVPGAGVPGVGVPGVGVPGVGVPGVGVPGAGV PGVGVPGVGVPGVGVPGVGVPGAGVPGVGVPGVGVPGVGVPGVGVPGAGVPGVGVPGVG VPGVGVPGVGVPGAGVPGVGVPGVGVPGVGVPGVGVPGAGVPGVGVPGVGVPGVGVPGV GVPGAGVPGVGVPGVGVPGVGVPGVGVPGAGVPGVGVPGVGVPGVGVPGVGVPGAGVPG VGVPGVGVPGVGVPGVGVPGAGY*DIQMTQSPSSLSASVGDRVTITCRASQDVNTAVAWYQQ KPGKAPKLLIYSASFLYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQHYTTPPTFGQGT KVEIKRTVAAPSVFIFPPSDEQLKSGTASWCLLNNFYPREAKVQWKVDNALQSGNSQESVTE QDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGECGY*SEQ ID NO: 63Trastuzumab-HC-V(80)\LC EVQLVESGGGLVQPGGSLRLSCAASGFNIKDTYIHWVRQAPGKGLEWVARIYPTNGYTRYADS VKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCSRWGGDGFYAMDYWGQGTLVTVSSASTK GPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSS VVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPK DTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQ DWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYP SDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHY TQKSLSLSPGKGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPG VGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVP GVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGV PGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVG VPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGV GVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPG VGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGY*D IQMTQSPSSLSASVGDRVTITCRASQDVNTAVAWYQQKPGKAPKLLIYSASFLYSGVPSRFSG SRSGTDFTLTISSLQPEDFATYYCQQHYTTPPTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGT ASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVY ACEVTHQGLSSPVTKSFNRGECGY*EVQLVESGGGLVQPGGSLRLSCAASGFNIKDTYIHWVR QAPGKGLEWVARIYPTNGYTRYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCSRWG GDGFYAMDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWN SGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTDocket No. 028193-0052-WO01HTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCWVDVSHEDPEVKFNWYVDGVEVHN AKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVY TLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTV DKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGKGVPGVGVPGVGVPGVGVPGVGVPGV GVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPG VGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVP GVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGV PGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVG VPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGV GVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPG VGVPGVGVPGVGVPGVGVPGVGY*DIQMTQSPSSLSASVGDRVTITCRASQDVNTAVAWYQQ KPGKAPKLLIYSASFLYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQHYTTPPTFGQGT KVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTE QDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGECGY*SEQ ID NO: 64Trastuzumab-HC-SISI(80) | LC EVQLVESGGGLVQPGGSLRLSCAASGFNIKDTYIHWVRQAPGKGLEWVARIYPTNGYTRYADS VKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCSRWGGDGFYAMDYWGQGTLVTVSSASTK GPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSS VVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPK DTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQ DWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYP SDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHY TQKSLSLSPGKGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPG SGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGIGVPG IGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIG VPGIGVPGIGVPGIGVPGIGVPGIGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVP GSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGV PGSGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVP GIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGY*DIQMTQSPSSLSASVGDRVTITCRAS QDVNTAVAWYQQKPGKAPKLLIYSASFLYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQ QHYTTPPTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASWCLLNNFYPREAKVQWKVDN ALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC*Docket No. 028193-0052-WO01EVQLVESGGGLVQPGGSLRLSCAASGFNIKDTYIHWVRQAPGKGLEWVARIYPTNGYTRYADS VKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCSRWGGDGFYAMDYWGQGTLVTVSSASTK GPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSS VVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPK DTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQ DWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYP SDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHY TQKSLSLSPGKGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPG SGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGIGVPG IGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIG VPGIGVPGIGVPGIGVPGIGVPGIGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVP GSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGV PGSGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVP GIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGY*DIQMTQSPSSLSASVGDRVTITCRAS QDVNTAVAWYQQKPGKAPKLLIYSASFLYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQ QHYTTPPTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDN ALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC*SEQ ID NO: 65Trastuzumab-HC\LC-SISI(80) EVQLVESGGGLVQPGGSLRLSCAASGFNIKDTYIHWVRQAPGKGLEWVARIYPTNGYTRYADS VKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCSRWGGDGFYAMDYWGQGTLVTVSSASTK GPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSS VVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPK DTLMISRTPEVTCVWDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQ DWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYP SDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHY TQKSLSLSPGK*DIQMTQSPSSLSASVGDRVTITCRASQDVNTAVAWYQQKPGKAPKLLIYSAS FLYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQHYTTPPTFGQGTKVEIKRTVAAPSVFI FPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTL TLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGECGVPGSGVPGSGVPGSGVPGSGVPGSG VPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGS GVPGSGVPGSGVPGSGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGI GVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGSGVPGSGVPGSDocket No. 028193-0052-WO01GVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPG SGVPGSGVPGSGVPGSGVPGSGVPGSGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGV PGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGY*E VQLVESGGGLVQPGGSLRLSCAASGFNIKDTYIHWVRQAPGKGLEWVARIYPTNGYTRYADSV KGRFTISADTSKNTAYLQMNSLRAEDTAVYYCSRWGGDGFYAMDYWGQGTLVTVSSASTKG PSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSV VTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKD TLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQD WLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPS DIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYT QKSLSLSPGK*DIQMTQSPSSLSASVGDRVTITCRASQDVNTAVAWYQQKPGKAPKLLIYSASF LYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQHYTTPPTFGQGTKVEIKRTVAAPSVFIF PPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTL SKADYEKHKVYACEVTHQGLSSPVTKSFNRGECGVPGSGVPGSGVPGSGVPGSGVPGSGVP GSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGV PGSGVPGSGVPGSGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGV PGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGSGVPGSGVPGSGV PGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSG VPGSGVPGSGVPGSGVPGSGVPGSGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGI GVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGY*SEQ ID NO: 66Trastuzumab-HC-SISI(80)\LC-SISI(80) EVQLVESGGGLVQPGGSLRLSCAASGFNIKDTYIHWVRQAPGKGLEWVARIYPTNGYTRYADS VKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCSRWGGDGFYAMDYWGQGTLVTVSSASTK GPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSS VVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPK DTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQ DWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYP SDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHY TQKSLSLSPGKGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPG SGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGIGVPG IGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIG VPGIGVPGIGVPGIGVPGIGVPGIGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPDocket No. 028193-0052-WO01GSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGV PGSGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVP GIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGY*DIQMTQSPSSLSASVGDRVTITCRAS QDVNTAVAWYQQKPGKAPKLLIYSASFLYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQ QHYTTPPTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDN ALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGECG VPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGS GVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGIGVPGIGVPGIGVPGIG VPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVP GIGVPGIGVPGIGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPG SGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGIGVPG IGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIG VPGIGVPGIGVPGIGVPGIGVPGIGY*EVQLVESGGGLVQPGGSLRLSCAASGFNIKDTYIHWVR QAPGKGLEWVARIYPTNGYTRYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCSRWG GDGFYAMDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWN SGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKT HTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHN AKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVY TLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTV DKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGKGVPGSGVPGSGVPGSGVPGSGVPGS GVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPG SGVPGSGVPGSGVPGSGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVP GIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGSGVPGSGVP GSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGV PGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGI GVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGY*DIQMTQSPSSLSASVGDRVTITCRASQDVNTAVAWYQQKPGKAPKLLIYSASFLYSGVPSRF SGSRSGTDFTLTISSLQPEDFATYYCQQHYTTPPTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKS GTASWCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKH KVYACEVTHQGLSSPVTKSFNRGECGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGS GVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPG SGVPGSGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGI GVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGSGVPGSGVPGSGVPGSGVPG SGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPDocket No. 028193-0052-WO01GSGVPGSGVPGSGVPGSGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGV PGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGY*SEQ ID NO: 67Trastuzumab-HC-ISIS(80) | LC EVQLVESGGGLVQPGGSLRLSCAASGFNIKDTYIHWVRQAPGKGLEWVARIYPTNGYTRYADS VKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCSRWGGDGFYAMDYWGQGTLVTVSSASTK GPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSS VVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPK DTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQ DWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYP SDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHY TQKSLSLSPGKGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGI GVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGSGVPGSGVPGSGVPG SGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVP GSGVPGSGVPGSGVPGSGVPGSGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGV PGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGSGVP GSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGV PGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGY*DIQMTQSPSSLSASVGDRVTITCRAS QDVNTAVAWYQQKPGKAPKLLIYSASFLYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQ QHYTTPPTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASWCLLNNFYPREAKVQWKVDN ALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC* EVQLVESGGGLVQPGGSLRLSCAASGFNIKDTYIHWVRQAPGKGLEWVARIYPTNGYTRYADS VKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCSRWGGDGFYAMDYWGQGTLVTVSSASTK GPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSS VVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPK DTLMISRTPEVTCVWDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQ DWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYP SDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHY TQKSLSLSPGKGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGI GVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGSGVPGSGVPGSGVPG SGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVP GSGVPGSGVPGSGVPGSGVPGSGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGV PGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGSGVPDocket No. 028193-0052-WO01GSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGV PGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGY*DIQMTQSPSSLSASVGDRVTITCRAS QDVNTAVAWYQQKPGKAPKLLIYSASFLYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQ QHYTTPPTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDN ALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC*SEQ ID NO: 68Trastuzumab-HC|LC-ISIS(80) EVQLVESGGGLVQPGGSLRLSCAASGFNIKDTYIHWVRQAPGKGLEWVARIYPTNGYTRYADS VKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCSRWGGDGFYAMDYWGQGTLVTVSSASTK GPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSS VVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPK DTLMISRTPEVTCVWDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQ DWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYP SDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHY TQKSLSLSPGK*DIQMTQSPSSLSASVGDRVTITCRASQDVNTAVAWYQQKPGKAPKLLIYSAS FLYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQHYTTPPTFGQGTKVEIKRTVAAPSVFI FPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTL TLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGECGVPGIGVPGIGVPGIGVPGIGVPGIGVPG IGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIG VPGIGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSG VPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGIGVPGIGVPGIGV PGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVP GIGVPGIGVPGIGVPGIGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPG SGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGY*E VQLVESGGGLVQPGGSLRLSCAASGFNIKDTYIHWVRQAPGKGLEWVARIYPTNGYTRYADSV KGRFTISADTSKNTAYLQMNSLRAEDTAVYYCSRWGGDGFYAMDYWGQGTLVTVSSASTKG PSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSV VTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKD TLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQD WLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPS DIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYT QKSLSLSPGK*DIQMTQSPSSLSASVGDRVTITCRASQDVNTAVAWYQQKPGKAPKLLIYSASF LYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQHYTTPPTFGQGTKVEIKRTVAAPSVFIFDocket No. 028193-0052-WO01PPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTL SKADYEKHKVYACEVTHQGLSSPVTKSFNRGECGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIG VPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVP GIGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVP GSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGIGVPGIGVPGIGVPG IGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIG VPGIGVPGIGVPGIGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGV PGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGY*SEQ ID NO: 69Trastuzumab-HC-ISIS(80)|LC-ISIS(80) EVQLVESGGGLVQPGGSLRLSCAASGFNIKDTYIHWVRQAPGKGLEWVARIYPTNGYTRYADS VKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCSRWGGDGFYAMDYWGQGTLVTVSSASTK GPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSS VVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPK DTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQ DWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYP SDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHY TQKSLSLSPGKGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGI GVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGSGVPGSGVPGSGVPG SGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVP GSGVPGSGVPGSGVPGSGVPGSGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGV PGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGSGVP GSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGV PGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGY*DIQMTQSPSSLSASVGDRVTITCRAS QDVNTAVAWYQQKPGKAPKLLIYSASFLYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQ QHYTTPPTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASWCLLNNFYPREAKVQWKVDN ALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGECG VPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVP GIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSG VPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGS GVPGSGVPGSGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGI GVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGSGVPGSGVPGSGVPG SGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPDocket No. 028193-0052-WO01GSGVPGSGVPGSGVPGSGVPGSGY*EVQLVESGGGLVQPGGSLRLSCAASGFNIKDTYIHWV RQAPGKGLEWVARIYPTNGYTRYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCSRW GGDGFYAMDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSW NSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCD KTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEV HNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQ VYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKL TVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGKGVPGIGVPGIGVPGIGVPGIGVPGIG VPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVP GIGVPGIGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVP GSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGIGVPGIGVP GIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGI GVPGIGVPGIGVPGIGVPGIGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSG VPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGS GY*DIQMTQSPSSLSASVGDRVTITCRASQDVNTAVAWYQQKPGKAPKLLIYSASFLYSGVPSR FSGSRSGTDFTLTISSLQPEDFATYYCQQHYTTPPTFGQGTKVEIKRTVAAPSVFIFPPSDEQLK SGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEK HKVYACEVTHQGLSSPVTKSFNRGECGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVP GIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPG SGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVP GSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGIGVPGIGVPGIGVPGIGVPGI GVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIG VPGIGVPGIGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSG VPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGY*SEQ ID NO: 70Trastuzumab-HC-IISS(80) | LC EVQLVESGGGLVQPGGSLRLSCAASGFNIKDTYIHWVRQAPGKGLEWVARIYPTNGYTRYADS VKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCSRWGGDGFYAMDYWGQGTLVTVSSASTK GPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSS VVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPK DTLMISRTPEVTCVWDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQ DWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYP SDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYDocket No. 028193-0052-WO01TQKSLSLSPGKGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGI GVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIG VPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVP GIGVPGIGVPGIGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPG SGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVP GSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGV PGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGY*DIQMTQSPSSLSASVGDRVTITCRAS QDVNTAVAWYQQKPGKAPKLLIYSASFLYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQ QHYTTPPTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASWCLLNNFYPREAKVQWKVDN ALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC* EVQLVESGGGLVQPGGSLRLSCAASGFNIKDTYIHWVRQAPGKGLEWVARIYPTNGYTRYADS VKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCSRWGGDGFYAMDYWGQGTLVTVSSASTK GPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSS VVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPK DTLMISRTPEVTCVWDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQ DWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYP SDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHY TQKSLSLSPGKGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGI GVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIG VPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVP GIGVPGIGVPGIGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPG SGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVP GSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGV PGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGY*DIQMTQSPSSLSASVGDRVTITCRAS QDVNTAVAWYQQKPGKAPKLLIYSASFLYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQ QHYTTPPTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDN ALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC*SEQ ID NO: 71Trastuzumab-HC\LC-IISS(80) EVQLVESGGGLVQPGGSLRLSCAASGFNIKDTYIHWVRQAPGKGLEWVARIYPTNGYTRYADS VKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCSRWGGDGFYAMDYWGQGTLVTVSSASTK GPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSS VVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDocket No. 028193-0052-WO01DTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQ DWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYP SDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHY TQKSLSLSPGK*DIQMTQSPSSLSASVGDRVTITCRASQDVNTAVAWYQQKPGKAPKLLIYSAS FLYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQHYTTPPTFGQGTKVEIKRTVAAPSVFI FPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTL TLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGECGVPGIGVPGIGVPGIGVPGIGVPGIGVPG IGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIG VPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVP GIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGSGVPGSGVPGSGVPGSGVPGSGV PGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSG VPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGS GVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGY*EV QLVESGGGLVQPGGSLRLSCAASGFNIKDTYIHWVRQAPGKGLEWVARIYPTNGYTRYADSVK GRFTISADTSKNTAYLQMNSLRAEDTAVYYCSRWGGDGFYAMDYWGQGTLVTVSSASTKGP SVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVV TVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDT LMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQD WLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPS DIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYT QKSLSLSPGK*DIQMTQSPSSLSASVGDRVTITCRASQDVNTAVAWYQQKPGKAPKLLIYSASF LYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQHYTTPPTFGQGTKVEIKRTVAAPSVFIF PPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTL SKADYEKHKVYACEVTHQGLSSPVTKSFNRGECGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIG VPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVP GIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGI GVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGSGVPGSGVPGSGVPGSGVPGSGVP GSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGV PGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSG VPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGY*SEQ ID NO: 72Trastuzumab-HC-IISS(80) | \LC-IISS(80)Docket No. 028193-0052-WO01EVQLVESGGGLVQPGGSLRLSCAASGFNIKDTYIHWVRQAPGKGLEWVARIYPTNGYTRYADS VKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCSRWGGDGFYAMDYWGQGTLVTVSSASTK GPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSS VVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPK DTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQ DWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYP SDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHY TQKSLSLSPGKGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGI GVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIG VPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVP GIGVPGIGVPGIGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPG SGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVP GSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGV PGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGY*DIQMTQSPSSLSASVGDRVTITCRAS QDVNTAVAWYQQKPGKAPKLLIYSASFLYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQ QHYTTPPTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDN ALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGECG VPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVP GIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGI GVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIG VPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGS GVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPG SGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVP GSGVPGSGVPGSGVPGSGVPGSGY*EVQLVESGGGLVQPGGSLRLSCAASGFNIKDTYIHWV RQAPGKGLEWVARIYPTNGYTRYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCSRW GGDGFYAMDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSW NSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCD KTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEV HNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQ VYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKL TVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGKGVPGIGVPGIGVPGIGVPGIGVPGIG VPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVP GIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGI GVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGSGVPGSGVPGSGVPGSGVPGDocket No. 028193-0052-WO01SGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVP GSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGV PGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGY*DIQMTQSPSSLSASVGDRVTITCRASQDVNTAVAWYQQKPGKAPKLLIYSASFLYSGVPSRF SGSRSGTDFTLTISSLQPEDFATYYCQQHYTTPPTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKS GTASWCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKH KVYACEVTHQGLSSPVTKSFNRGECGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGI GVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIG VPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVP GIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGS GVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPG SGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVP GSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGY*SEQ ID NO: 73Trastuzumab-HC-SSII(80) | LC EVQLVESGGGLVQPGGSLRLSCAASGFNIKDTYIHWVRQAPGKGLEWVARIYPTNGYTRYADS VKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCSRWGGDGFYAMDYWGQGTLVTVSSASTK GPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSS VVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPK DTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGKGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPG SGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVP GSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGV PGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGIGVPGIGVPGIGVPGIGVPGIGVPGI GVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIG VPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVP GIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGY*DIQMTQSPSSLSASVGDRVTITCRAS QDVNTAVAWYQQKPGKAPKLLIYSASFLYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQ QHYTTPPTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDN ALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC* EVQLVESGGGLVQPGGSLRLSCAASGFNIKDTYIHWVRQAPGKGLEWVARIYPTNGYTRYADSDocket No. 028193-0052-WO01VKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCSRWGGDGFYAMDYWGQGTLVTVSSASTK GPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSS VVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPK DTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGKGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPG SGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVP GSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGV PGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGIGVPGIGVPGIGVPGIGVPGIGVPGI GVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIG VPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVP GIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGY*DIQMTQSPSSLSASVGDRVTITCRAS QDVNTAVAWYQQKPGKAPKLLIYSASFLYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQ QHYTTPPTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASWCLLNNFYPREAKVQWKVDN ALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC*SEQ ID NO: 74Trastuzumab-HC\LC-SSII(80) EVQLVESGGGLVQPGGSLRLSCAASGFNIKDTYIHWVRQAPGKGLEWVARIYPTNGYTRYADS VKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCSRWGGDGFYAMDYWGQGTLVTVSSASTK GPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSS VVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPK DTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQ DWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYP SDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHY TQKSLSLSPGK*DIQMTQSPSSLSASVGDRVTITCRASQDVNTAVAWYQQKPGKAPKLLIYSAS FLYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQHYTTPPTFGQGTKVEIKRTVAAPSVFI FPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTL TLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGECGVPGSGVPGSGVPGSGVPGSGVPGSG VPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGS GVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPG SGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVP GIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIDocket No. 028193-0052-WO01GVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIG VPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGY* EVQLVESGGGLVQPGGSLRLSCAASGFNIKDTYIHWVRQAPGKGLEWVARIYPTNGYTRYADS VKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCSRWGGDGFYAMDYWGQGTLVTVSSASTK GPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSS VVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPK DTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQ DWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYP SDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHY TQKSLSLSPGK*DIQMTQSPSSLSASVGDRVTITCRASQDVNTAVAWYQQKPGKAPKLLIYSAS FLYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQHYTTPPTFGQGTKVEIKRTVAAPSVFI FPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTL TLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGECGVPGSGVPGSGVPGSGVPGSGVPGSG VPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGS GVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPG SGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVP GIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGI GVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIG VPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGY*SEQ ID NO: 75Trastuzumab-HC-SSII(80)\LC-SSII(80) EVQLVESGGGLVQPGGSLRLSCAASGFNIKDTYIHWVRQAPGKGLEWVARIYPTNGYTRYADS VKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCSRWGGDGFYAMDYWGQGTLVTVSSASTK GPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSS VVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPK DTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGKGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPG SGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVP GSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGV PGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGIGVPGIGVPGIGVPGIGVPGIGVPGI GVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGDocket No. 028193-0052-WO01VPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVP GIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGY*DIQMTQSPSSLSASVGDRVTITCRAS QDVNTAVAWYQQKPGKAPKLLIYSASFLYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQ QHYTTPPTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDN ALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGECG VPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGS GVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPG SGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVP GSGVPGSGVPGSGVPGSGVPGSGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGV PGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVP GIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGI GVPGIGVPGIGVPGIGVPGIGVPGIGY*EVQLVESGGGLVQPGGSLRLSCAASGFNIKDTYIHWV RQAPGKGLEWVARIYPTNGYTRYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCSRW GGDGFYAMDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSW NSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCD KTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEV HNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQ VYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKL TVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGKGVPGSGVPGSGVPGSGVPGSGVP GSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGV PGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSG VPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGS GVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIG VPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVP GIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGI GY*DIQMTQSPSSLSASVGDRVTITCRASQDVNTAVAWYQQKPGKAPKLLIYSASFLYSGVPSR FSGSRSGTDFTLTISSLQPEDFATYYCQQHYTTPPTFGQGTKVEIKRTVAAPSVFIFPPSDEQLK SGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEK HKVYACEVTHQGLSSPVTKSFNRGECGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPG SGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVP GSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGV PGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGIGVPGIGV PGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPDocket No. 028193-0052-WO01GIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGI GVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGY*SEQ ID NO: 76Trastuzumab-HC-SISISI(120)\LC EVQLVESGGGLVQPGGSLRLSCAASGFNIKDTYIHWVRQAPGKGLEWVARIYPTNGYTRYADS VKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCSRWGGDGFYAMDYWGQGTLVTVSSASTK GPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSS VVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPK DTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGKGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPG SGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGIGVPG IGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIG VPGIGVPGIGVPGIGVPGIGVPGIGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVP GSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGV PGSGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVP GIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGSGVPGSGVPGSGVPGSGVPGSGV PGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSG VPGSGVPGSGVPGSGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIG VPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGY*DIQMTQSPSSLSAS VGDRVTITCRASQDVNTAVAWYQQKPGKAPKLLIYSASFLYSGVPSRFSGSRSGTDFTLTISSL QPEDFATYYCQQHYTTPPTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPR EAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSP VTKSFNRGEC*EVQLVESGGGLVQPGGSLRLSCAASGFNIKDTYIHWVRQAPGKGLEWVARIY PTNGYTRYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCSRWGGDGFYAMDYWGQG TLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAV LQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLG GPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNST YRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQ VSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFS CSVM H EALH N HYTQKSLSLSPGKGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGV PGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGDocket No. 028193-0052-WO01VPGSGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGV PGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGSGVPGSGVPGSGVPGSGVPGSG VPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGS GVPGSGVPGSGVPGSGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGI GVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGSGVPGSGVPGS GVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPG SGVPGSGVPGSGVPGSGVPGSGVPGSGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGV PGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGY*D IQMTQSPSSLSASVGDRVTITCRASQDVNTAVAWYQQKPGKAPKLLIYSASFLYSGVPSRFSG SRSGTDFTLTISSLQPEDFATYYCQQHYTTPPTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGT ASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVY ACEVTHQGLSSPVTKSFNRGEC*SEQ ID NO: 77Trastuzumab-HC\LC-SISISI(120) EVQLVESGGGLVQPGGSLRLSCAASGFNIKDTYIHWVRQAPGKGLEWVARIYPTNGYTRYADS VKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCSRWGGDGFYAMDYWGQGTLVTVSSASTK GPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSS VVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPK DTLMISRTPEVTCVWDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQ DWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYP SDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHY TQKSLSLSPGK*DIQMTQSPSSLSASVGDRVTITCRASQDVNTAVAWYQQKPGKAPKLLIYSAS FLYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQHYTTPPTFGQGTKVEIKRTVAAPSVFI FPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTL TLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGECGVPGSGVPGSGVPGSGVPGSGVPGSG VPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGS GVPGSGVPGSGVPGSGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGI GVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGSGVPGSGVPGS GVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPG SGVPGSGVPGSGVPGSGVPGSGVPGSGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGV PGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVP GSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGV PGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGIGVPGIGVPGIGVPGIGVPDocket No. 028193-0052-WO01GIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGI GVPGIGVPGIGY*EVQLVESGGGLVQPGGSLRLSCAASGFNIKDTYIHWVRQAPGKGLEWVARI YPTNGYTRYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCSRWGGDGFYAMDYWGQ GTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPA VLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLG GPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNST YRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQ VSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFS CSVMHEALHNHYTQKSLSLSPGK*DIQMTQSPSSLSASVGDRVTITCRASQDVNTAVAWYQQK PGKAPKLLIYSASFLYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQHYTTPPTFGQGTK VEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQ DSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGECGVPGSGVPGSGVPG SGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVP GSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIG VPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVP GSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGV PGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGIGVPGIGVPGIGVPGIGVP GIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGI GVPGIGVPGIGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGS GVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGIGVPGI GVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIG VPGIGVPGIGVPGIGVPGIGVPGIGY*SEQ ID NO: 78Trastuzumab-HC-SISISI(120)\LC-SISISI(120) EVQLVESGGGLVQPGGSLRLSCAASGFNIKDTYIHWVRQAPGKGLEWVARIYPTNGYTRYADS VKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCSRWGGDGFYAMDYWGQGTLVTVSSASTK GPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSS VVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPK DTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQ DWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYP SDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHY TQKSLSLSPGKGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPG SGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGIGVPGDocket No. 028193-0052-WO01IGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIG VPGIGVPGIGVPGIGVPGIGVPGIGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVP GSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGV PGSGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVP GIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGSGVPGSGVPGSGVPGSGVPGSGV PGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSG VPGSGVPGSGVPGSGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIG VPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGY*DIQMTQSPSSLSAS VGDRVTITCRASQDVNTAVAWYQQKPGKAPKLLIYSASFLYSGVPSRFSGSRSGTDFTLTISSL QPEDFATYYCQQHYTTPPTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPR EAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSP VTKSFNRGECGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGS GVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGIGVPGI GVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIG VPGIGVPGIGVPGIGVPGIGVPGIGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVP GSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGV PGSGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVP GIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGSGVPGSGVPGSGVPGSGVPGSGV PGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSG VPGSGVPGSGVPGSGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIG VPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGY*EVQLVESGGGLVQP GGSLRLSCAASGFNIKDTYIHWVRQAPGKGLEWVARIYPTNGYTRYADSVKGRFTISADTSKNT AYLQMNSLRAEDTAVYYCSRWGGDGFYAMDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTS GGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYI CNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCV VVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKV SNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQP ENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGKGV PGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSG VPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGIGVPGIGVPGIGVPGIGVP GIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGI GVPGIGVPGIGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGS GVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGIGVPGI GVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGDocket No. 028193-0052-WO01VPGIGVPGIGVPGIGVPGIGVPGIGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVP GSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGV PGSGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVP GIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGY*DIQMTQSPSSLSASVGDRVTITCRAS QDVNTAVAWYQQKPGKAPKLLIYSASFLYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQ QHYTTPPTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDN ALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGECG VPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGS GVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGIGVPGIGVPGIGVPGIG VPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVP GIGVPGIGVPGIGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPG SGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGIGVPG IGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIG VPGIGVPGIGVPGIGVPGIGVPGIGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVP GSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGV PGSGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVP GIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGY*SEQ ID NO: 79Dulaglutide-SI40 HGEGTFTSDVSSYLEEQAAKEFIAWLVKGGGGGGGSGGGGSGGGGSAESKYGPPCPPCPAP EAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREE QFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEE MTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQE G N VFSCSVM H EALH N H YTQKSLSLSLGG VPGSG VPGSG VPGSGVPGSGVPGSGVPGSGVPG SGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVP GSGVPGSGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVP GIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGYSEQ ID NO: 80Dulaglutide-ISIS80 HGEGTFTSDVSSYLEEQAAKEFIAWLVKGGGGGGGSGGGGSGGGGSAESKYGPPCPPCPAP EAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREE QFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEDocket No. 028193-0052-WO01MTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQE GNVFSCSVMHEALHNHYTQKSLSLSLGGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGV PGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVP GSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGV PGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGIGVPGIGVPGIGVPGIGVP GIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGI GVPGIGVPGIGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGS GVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGYSEQ ID NO: 81Dulaglutide-SSII80 HGEGTFTSDVSSYLEEQAAKEFIAWLVKGGGGGGGSGGGGSGGGGSAESKYGPPCPPCPAP EAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREE QFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEE MTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQE G N VFSCSVM H EALH N H YTQKSLSLSLGG VPGSG VPGSG VPGSGVPGSGVPGSGVPGSGVPG SGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVP GSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGV PGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGIGVPGIGV PGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVP GIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGI GVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGYSEQ ID NO: 82Dulaglutide-IISS80 HGEGTFTSDVSSYLEEQAAKEFIAWLVKGGGGGGGSGGGGSGGGGSAESKYGPPCPPCPAP EAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREE QFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEE MTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQE GNVFSCSVMHEALHNHYTQKSLSLSLGGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGV PGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVP GIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGI GVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVP GSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVDocket No. 028193-0052-WO01PGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSG VPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGYSEQ ID NO: 83Dulaglutide-SISISI120 HGEGTFTSDVSSYLEEQAAKEFIAWLVKGGGGGGGSGGGGSGGGGSAESKYGPPCPPCPAP EAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREE QFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEE MTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQE G N VFSCSVM H EALH N H YTQKSLSLSLGG VPGSG VPGSG VPGSGVPGSGVPGSGVPGSGVPG SGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVP GSGVPGSGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVP GIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGSGVPGSGVPGSGVPGSGV PGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSG VPGSGVPGSGVPGSGVPGSGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGI GVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGSGVPGS GVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPG SGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGV PGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVP GIGYSEQ ID NO: 84Dulaglutide-SISI80 HGEGTFTSDVSSYLEEQAAKEFIAWLVKGGGGGGGSGGGGSGGGGSAESKYGPPCPPCPAP EAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREE QFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEE MTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQE G N VFSCSVM H EALH N H YTQKSLSLSLGG VPGSG VPGSG VPGSGVPGSGVPGSGVPGSGVPG SGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVP GSGVPGSGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVP GIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGSGVPGSGVPGSGVPGSGV PGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSG VPGSGVPGSGVPGSGVPGSGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGI GVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGYDocket No. 028193-0052-WO01SEQ ID NO: 85Dulaglutide-Slalt80 HGEGTFTSDVSSYLEEQAAKEFIAWLVKGGGGGGGSGGGGSGGGGSAESKYGPPCPPCPAP EAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREE QFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEE MTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQE GNVFSCSVMHEALHNHYTQKSLSLSLGGVPGSGVPGIGVPGSGVPGIGVPGSGVPGIGVPGS GVPGIGVPGSGVPGIGVPGSGVPGIGVPGSGVPGIGVPGSGVPGIGVPGSGVPGIGVPGSGVP GIGVPGSGVPGIGVPGSGVPGIGVPGSGVPGIGVPGSGVPGIGVPGSGVPGIGVPGSGVPGIG VPGSGVPGIGVPGSGVPGIGVPGSGVPGIGVPGSGVPGIGVPGSGVPGIGVPGSGVPGIGVPG SGVPGIGVPGSGVPGIGVPGSGVPGIGVPGSGVPGIGVPGSGVPGIGVPGSGVPGIGVPGSGV PGIGVPGSGVPGIGVPGSGVPGIGVPGSGVPGIGVPGSGVPGIGVPGSGVPGIGVPGSGVPGI GVPGSGVPGIGVPGSGVPGIGVPGSGVPGIGVPGSGVPGIGVPGSGVPGIGYSEQ ID NO: 86Dulaglutide-V40 HGEGTFTSDVSSYLEEQAAKEFIAWLVKGGGGGGGSGGGGSGGGGSAESKYGPPCPPCPAP EAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREE QFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEE MTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQE G N VFSCSVM H EALH N H YTQKSLSLSLGG VPG VGVPGVGVPG VGVPGVGVPG VGVPGVGVPG VGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVP GVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGV PGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGYSEQ ID NO: 87Dulaglutide-V80 HGEGTFTSDVSSYLEEQAAKEFIAWLVKGGGGGGGSGGGGSGGGGSAESKYGPPCPPCPAP EAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREE QFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEE MTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQE G N VFSCSVM H EALH N H YTQKSLSLSLGG VPG VGVPGVGVPG VGVPGVGVPG VGVPGVGVPG VGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPDocket No. 028193-0052-WO01GVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGV PGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVG VPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGV GVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPG VGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVP GVGVPGVGVPGVGYSEQ ID NO: 88Dulaglutide- V 120 HGEGTFTSDVSSYLEEQAAKEFIAWLVKGGGGGGGSGGGGSGGGGSAESKYGPPCPPCPAP EAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREE QFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEE MTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQE G N VFSCSVM H EALH N H YTQKSLSLSLGG VPG VGVPGVGVPG VGVPGVGVPG VGVPGVGVPG VGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVP GVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGV PGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVG VPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGV GVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPG VGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVP GVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGV PGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVG VPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGV GVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGYSEQ ID NO: 89Dulaglutide-(A1V1)80 HGEGTFTSDVSSYLEEQAAKEFIAWLVKGGGGGGGSGGGGSGGGGSAESKYGPPCPPCPAP EAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREE QFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEE MTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQE G N VFSCSVM H EALH N H YTQKSLSLSLGG VPG VGVPGAG VPG VGVPGAGVPGVGVPGAGVPG VGVPGAGVPGVGVPGAGVPGVGVPGAGVPGVGVPGAGVPGVGVPGAGVPGVGVPGAGVP GVGVPGAGVPGVGVPGAGVPGVGVPGAGVPGVGVPGAGVPGVGVPGAGVPGVGVPGAGVDocket No. 028193-0052-WO01PGVGVPGAGVPGVGVPGAGVPGVGVPGAGVPGVGVPGAGVPGVGVPGAGVPGVGVPGAG VPGVGVPGAGVPGVGVPGAGVPGVGVPGAGVPGVGVPGAGVPGVGVPGAGVPGVGVPGA GVPGVGVPGAGVPGVGVPGAGVPGVGVPGAGVPGVGVPGAGVPGVGVPGAGVPGVGVPG AGVPGVGVPGAGVPGVGVPGAGVPGVGVPGAGVPGVGVPGAGVPGVGVPGAGVPGVGVP GAGVPGVGVPGAGYSEQ ID NO: 90Dulaglutide-(A1V4)80 HGEGTFTSDVSSYLEEQAAKEFIAWLVKGGGGGGGSGGGGSGGGGSAESKYGPPCPPCPAP EAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREE QFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEE MTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQE G N VFSCSVM H EALH N H YTQKSLSLSLGG VPG VG VPG VG VPG VGVPGVGVPGAGVPGVGVPG VGVPGVGVPGVGVPGAGVPGVGVPGVGVPGVGVPGVGVPGAGVPGVGVPGVGVPGVGVP GVGVPGAGVPGVGVPGVGVPGVGVPGVGVPGAGVPGVGVPGVGVPGVGVPGVGVPGAGV PGVGVPGVGVPGVGVPGVGVPGAGVPGVGVPGVGVPGVGVPGVGVPGAGVPGVGVPGVG VPGVGVPGVGVPGAGVPGVGVPGVGVPGVGVPGVGVPGAGVPGVGVPGVGVPGVGVPGV GVPGAGVPGVGVPGVGVPGVGVPGVGVPGAGVPGVGVPGVGVPGVGVPGVGVPGAGVPG VGVPGVGVPGVGVPGVGVPGAGVPGVGVPGVGVPGVGVPGVGVPGAGVPGVGVPGVGVP GVGVPGVGVPGAGYSEQ ID NO: 91Dulaglutide-S80 HGEGTFTSDVSSYLEEQAAKEFIAWLVKGGGGGGGSGGGGSGGGGSAESKYGPPCPPCPAP EAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREE QFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEE MTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQE G N VFSCSVM H EALH N H YTQKSLSLSLGG VPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPG SGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVP GSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGV PGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSG VPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGS GVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGDocket No. 028193-0052-WO01SGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVP GSGVPGSGVPGSGYSEQ ID NO: 92Dulaglutide-S 120 HGEGTFTSDVSSYLEEQAAKEFIAWLVKGGGGGGGSGGGGSGGGGSAESKYGPPCPPCPAP EAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREE QFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEE MTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQE G N VFSCSVM H EALH N H YTQKSLSLSLGG VPGSG VPGSG VPGSGVPGSGVPGSGVPGSGVPG SGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVP GSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGV PGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSG VPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGS GVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPG SGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVP GSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGV PGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSG VPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGS GVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGYSEQ ID NO: 93Dulaglutide-A80 HGEGTFTSDVSSYLEEQAAKEFIAWLVKGGGGGGGSGGGGSGGGGSAESKYGPPCPPCPAP EAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREE QFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEE MTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQE G N VFSCSVM H EALH N H YTQKSLSLSLGG VPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPG AGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVP GAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGV PGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAG VPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGA GVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGDocket No. 028193-0052-WO01AGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVP GAGVPGAGVPGAGYSEQ ID NO: 94Dulaglutide-A 120 HGEGTFTSDVSSYLEEQAAKEFIAWLVKGGGGGGGSGGGGSGGGGSAESKYGPPCPPCPAP EAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREE QFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEE MTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQE G N VFSCSVM H EALH N H YTQKSLSLSLGG VPGAG VPGAG VPGAGVPGAGVPGAGVPGAGVPG AGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVP GAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGV PGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAG VPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGA GVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPG AGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVP GAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGV PGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAG VPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGA GVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGYSEQ ID NO: 95Mouse Light Chain - IVVI80 DIVMTQSPSSFSVSLGDRVTITCKASEDIYNRLAWYQQKPGNAPRLLIFGATSLETGVPSRFSG SGSGKDYTLSITSLATEDVATYYCQQYWTTPWTFGGGTKLEIKRADAAPTVSIFPPSSEQLTSG GASVVCFLNNFYPKDINVKWKIDGSERQNGVLNSWTDQDSKDSTYSMSSTLTLTKDEYERHN SYTCEATHKTSTSPIVKSFNRNECGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGV PGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGVGVP GVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGV PGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVG VPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGV GVPGVGVPGVGVPGVGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGI GVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGYDocket No. 028193-0052-WO01SEQ ID NO: 96Mouse Light Chain – V40DIVMTQSPSSFSVSLGDRVTITCKASEDIYNRLAWYQQKPGNAPRLLIFGATSLETGVPSRFSG SGSGKDYTLSITSLATEDVATYYCQQYWTTPWTFGGGTKLEIKRADAAPTVSIFPPSSEQLTSG GASVVCFLNNFYPKDINVKWKIDGSERQNGVLNSWTDQDSKDSTYSMSSTLTLTKDEYERHN SYTCEATHKTSTSPIVKSFNRNECGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGV PGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVG VPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGV GVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGYSEQ ID NO: 97Mouse Heavy Chain – V40-mCherryEVKLVESGGDLVKPGGSLKLSCAASAFTFSSYAMSWVRQTPEKRLEWVATISDGDSYTYYPD NVKGRFTISRDNAKNNLYLQMSHLKSEDTAMYYCTRDGASWDGTWFAYWGQGTLVTVSSGA KTTAPSVYPLAPVCGDTTGSSVTLGCLVKGYFPEPVTLTWNSGSLSSGVHTFPAVLQSDLYTL SSSVTVTSSTWPSQSITCNVAHPASSTKVDKKIEPRGPTIKPCPPCKCPAPNLLGGPSVFIFPPK IKDVLMISLSPIVTCVVVDVSEDDPDVQISWFVNNVEVHTAQTQTHREDYNSTLRVVSALPIQHQ DWMSGKEFKCKVNNKDLPAPIERTISKPKGSVRAPQVYVLPPPEEEMTKKQVTLTCMVTDFMP EDIYVEWTNNGKTELNYKNTEPVLDSDGSYFMYSKLRVEKKNWVERNSYSCSWHEGLHNHH TTKSFSRTPGKGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPG VGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVP GVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGV PGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVSKGDEDNMAIIKEFMRFKVHMEGSVN GHEFEIEGEGEGRPYEGTQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSF PEGFKWERVMNFEDGGWTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSE RMYPEDGALKGEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVE QYERAEGRHSTGGMDELYKSGYSEQ ID NO: 98Mouse Light Chain – S80-mCherryDIVMTQSPSSFSVSLGDRVTITCKASEDIYNRLAWYQQKPGNAPRLLIFGATSLETGVPSRFSGSGSGKDYTLSITSLATEDVATYYCQQYWTTPWTFGGGTKLEIKRADAAPTVSIFPPSSEQLTSGGASVVCFLNNFYPKDINVKWKIDGSERQNGVLNSWTDQDSKDSTYSMSSTLTLTKDEYERHNSYTCEATH KTSTSPI VKSFN RN ECGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVDocket No. 028193-0052-WO01PGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSG VPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGS GVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPG SGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVP GSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGV PGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSG VPGSGVPGSGVSKGDEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGTQTAKLKV TKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGVVTVTQD SSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKGEIKQRLKLKDGGH YDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERAEGRHSTGGMDELYKSGYSEQ ID NO: 99Mouse Light Chain - V40-mCherry DIVMTQSPSSFSVSLGDRVTITCKASEDIYNRLAWYQQKPGNAPRLLIFGATSLETGVPSRFSG SGSGKDYTLSITSI-ATEDVATYYCQQYWTTPWTFGGGTKLEIKRADAAPTVSIFPPSSEQLTSG GASVVCFLNNFYPKDINVKWKIDGSERQNGVLNSWTDQDSKDSTYSMSSTLTLTKDEYERHN SYTCEATHKTSTSPIVKSFNRNECGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGV PGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVG VPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGV GVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVSKGDEDNMAIIKEF MRFKVHMEGSVNGHEFEIEGEGEGRPYEGTQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYV KHPADIPDYLKLSFPEGFKWERVMNFEDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVM QKKTMGWEASSERMYPEDGALKGEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIK LDITSHNEDYTIVEQYERAEGRHSTGGMDELYKSGYSEQ ID NO: 100Mouse Heavy Chain - SISI80 VKLVESGGDLVKPGGSLKLSCAASAFTFSSYAMSWVRQTPEKRLEWVATISDGDSYTYYPDN VKGRFTISRDNAKNNLYLQMSHLKSEDTAMYYCTRDGASWDGTWFAYWGQGTLVTVSSGAK TTAPSVYPLAPVCGDTTGSSVTLGCLVKGYFPEPVTLTWNSGSLSSGVHTFPAVLQSDLYTLS SSVTVTSSTWPSQSITCNVAHPASSTKVDKKIEPRGPTIKPCPPCKCPAPNLLGGPSVFIFPPKI KDVLMISLSPIVTCVVVDVSEDDPDVQISWFVNNVEVHTAQTQTHREDYNSTLRVVSALPIQHQ DWMSGKEFKCKVNNKDLPAPIERTISKPKGSVRAPQVYVLPPPEEEMTKKQVTLTCMVTDFMP EDIYVEWTNNGKTELNYKNTEPVLDSDGSYFMYSKLRVEKKNWVERNSYSCSVVHEGLHNHHDocket No. 028193-0052-WO01TTKSFSRTPGKGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPG SGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGIGVPG IGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIG VPGIGVPGIGVPGIGVPGIGVPGIGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVP GSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGV PGSGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVP GIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGYSEQ ID NO: 101Mouse Light Chain - V80 DIVMTQSPSSFSVSLGDRVTITCKASEDIYNRLAWYQQKPGNAPRLLIFGATSLETGVPSRFSG SGSGKDYTLSITSLATEDVATYYCQQYWTTPWTFGGGTKLEIKRADAAPTVSIFPPSSEQLTSG GASVVCFLNNFYPKDINVKWKIDGSERQNGVLNSWTDQDSKDSTYSMSSTLTLTKDEYERHN SYTCEATHKTSTSPIVKSFNRNECGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGV PGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVG VPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGV GVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPG VGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVP GVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGV PGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVG VPGVGVPGVGYSEQ ID NO: 102Mouse Light Chain - SISI80 DIVMTQSPSSFSVSLGDRVTITCKASEDIYNRLAWYQQKPGNAPRLLIFGATSLETGVPSRFSG SGSGKDYTLSITSLATEDVATYYCQQYWTTPWTFGGGTKLEIKRADAAPTVSIFPPSSEQLTSG GASVVCFLNNFYPKDINVKWKIDGSERQNGVLNSWTDQDSKDSTYSMSSTLTLTKDEYERHN SYTCEATH KTSTSPI VKSFN RN ECGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGV PGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSG VPGSGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGV PGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGSGVPGSGVPGSGVPGSGVPGSG VPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGS GVPGSGVPGSGVPGSGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGI GVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGYDocket No. 028193-0052-WO01SEQ ID NO: 103Mouse Light Chain - VI40-mCherry DIVMTQSPSSFSVSLGDRVTITCKASEDIYNRLAWYQQKPGNAPRLLIFGATSLETGVPSRFSG SGSGKDYTLSITSLATEDVATYYCQQYWTTPWTFGGGTKLEIKRADAAPTVSIFPPSSEQLTSG GASVVCFLNNFYPKDINVKWKIDGSERQNGVLNSWTDQDSKDSTYSMSSTLTLTKDEYERHN SYTCEATHKTSTSPIVKSFNRNECGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGV PGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVG VPGVGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGV PGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVSKGDEDNMAIIKEFMRFKVHMEGSV NGHEFEIEGEGEGRPYEGTQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKL SFPEGFKWERVMNFEDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEAS SERMYPEDGALKGEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTI VEQYERAEGRHSTGGMDELYKSGYSEQ ID NO: 104Mouse Heavy Chain - IVVI80 EVKLVESGGDLVKPGGSLKLSCAASAFTFSSYAMSWVRQTPEKRLEWVATISDGDSYTYYPD NVKGRFTISRDNAKNNLYLQMSHLKSEDTAMYYCTRDGASWDGTWFAYWGQGTLVTVSSGA KTTAPSVYPLAPVCGDTTGSSVTLGCLVKGYFPEPVTLTWNSGSLSSGVHTFPAVLQSDLYTL SSSVTVTSSTWPSQSITCNVAHPASSTKVDKKIEPRGPTIKPCPPCKCPAPNLLGGPSVFIFPPK IKDVLMISLSPIVTCVVVDVSEDDPDVQISWFVNNVEVHTAQTQTHREDYNSTLRVVSALPIQHQ DWMSGKEFKCKVNNKDLPAPIERTISKPKGSVRAPQVYVLPPPEEEMTKKQVTLTCMVTDFMP EDIYVEWTNNGKTELNYKNTEPVLDSDGSYFMYSKLRVEKKNWVERNSYSCSVVHEGLHNHH TTKSFSRTPGKGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGI GVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGVGVPGVGVPGVGVPG VGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVP GVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGV PGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVG VPGVGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGV PGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGVPGIGYSEQ ID NO: 105Mouse Heavy Chain - A80-mCherryDocket No. 028193-0052-WO01EVKLVESGGDLVKPGGSLKLSCAASAFTFSSYAMSWVRQTPEKRLEWVATISDGDSYTYYPD NVKGRFTISRDNAKNNLYLQMSHLKSEDTAMYYCTRDGASWDGTWFAYWGQGTLVTVSSGA KTTAPSVYPLAPVCGDTTGSSVTLGCLVKGYFPEPVTLTWNSGSLSSGVHTFPAVLQSDLYTL SSSVTVTSSTWPSQSITCNVAHPASSTKVDKKIEPRGPTIKPCPPCKCPAPNLLGGPSVFIFPPK IKDVLMISLSPIVTCVVVDVSEDDPDVQISWFVNNVEVHTAQTQTHREDYNSTLRVVSALPIQHQ DWMSGKEFKCKVNNKDLPAPIERTISKPKGSVRAPQVYVLPPPEEEMTKKQVTLTCMVTDFMP EDIYVEWTNNGKTELNYKNTEPVLDSDGSYFMYSKLRVEKKNWVERNSYSCSWHEGLHNHH TTKSFSRTPGKGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPG AGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVP GAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGV PGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAG VPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGA GVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPG AGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVS KGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGTQTAKLKVTKGGPLPFAWDIL SPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGVVTVTQDSSLQDGEFIYKVKL RGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKGEIKQRLKLKDGGHYDAEVKTTYKAKK PVQLPGAYNVNIKLDITSHNEDYTIVEQYERAEGRHSTGGMDELYKGYSEQ ID NO: 106Mouse Heavy Chain - S80-mCherry EVKLVESGGDLVKPGGSLKLSCAASAFTFSSYAMSWVRQTPEKRLEWVATISDGDSYTYYPD NVKGRFTISRDNAKNNLYLQMSHLKSEDTAMYYCTRDGASWDGTWFAYWGQGTLVTVSSGA KTTAPSVYPLAPVCGDTTGSSVTLGCLVKGYFPEPVTLTWNSGSLSSGVHTFPAVLQSDLYTL SSSVTVTSSTWPSQSITCNVAHPASSTKVDKKIEPRGPTIKPCPPCKCPAPNLLGGPSVFIFPPK IKDVLMISLSPIVTCVVVDVSEDDPDVQISWFVNNVEVHTAQTQTHREDYNSTLRVVSALPIQHQ DWMSGKEFKCKVNNKDLPAPIERTISKPKGSVRAPQVYVLPPPEEEMTKKQVTLTCMVTDFMP EDIYVEWTNNGKTELNYKNTEPVLDSDGSYFMYSKLRVEKKNWVERNSYSCSVVHEGLHNHH TTKSFSRTPGKGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPG SGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVP GSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGV PGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSG VPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGS GVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGDocket No. 028193-0052-WO01SGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVPGSGVS KGDEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGTQTAKLKVTKGGPLPFAWDIL SPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGVVTVTQDSSLQDGEFIYKVKL RGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKGEIKQRLKLKDGGHYDAEVKTTYKAKK PVQLPGAYNVNIKLDITSHNEDYTIVEQYERAEGRHSTGGMDELYKSGYSEQ ID NO: 107Mouse Heavy Chain - V40 EVKLVESGGDLVKPGGSLKLSCAASAFTFSSYAMSWVRQTPEKRLEWVATISDGDSYTYYPD NVKGRFTISRDNAKNNLYLQMSHLKSEDTAMYYCTRDGASWDGTWFAYWGQGTLVTVSSGA KTTAPSVYPLAPVCGDTTGSSVTLGCLVKGYFPEPVTLTWNSGSLSSGVHTFPAVLQSDLYTL SSSVTVTSSTWPSQSITCNVAHPASSTKVDKKIEPRGPTIKPCPPCKCPAPNLLGGPSVFIFPPK IKDVLMISLSPIVTCVVVDVSEDDPDVQISWFVNNVEVHTAQTQTHREDYNSTLRVVSALPIQHQ DWMSGKEFKCKVNNKDLPAPIERTISKPKGSVRAPQVYVLPPPEEEMTKKQVTLTCMVTDFMP EDIYVEWTNNGKTELNYKNTEPVLDSDGSYFMYSKLRVEKKNWVERNSYSCSVVHEGLHNHH TTKSFSRTPGKGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPG VGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVP GVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGV PGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGYSEQ ID NO: 108Mouse Light Chain - A80-mCherry DIVMTQSPSSFSVSLGDRVTITCKASEDIYNRLAWYQQKPGNAPRLLIFGATSLETGVPSRFSG SGSGKDYTLSITSLATEDVATYYCQQYWTTPWTFGGGTKLEIKRADAAPTVSIFPPSSEQLTSG GASVVCFLNNFYPKDINVKWKIDGSERQNGVLNSWTDQDSKDSTYSMSSTLTLTKDEYERHN SYTCEATHKTSTSPIVKSFNRNECGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGV PGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAG VPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGA GVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPG AGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVP GAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGV PGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAGVPGAG VPGAGVPGAGVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGTQTAKLKV TKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGVVTVTQDDocket No. 028193-0052-WO01SSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKGEIKQRLKLKDGGH YDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERAEGRHSTGGMDELYKGYSEQ ID NO: 109Mouse Heavy Chain - V80 EVKLVESGGDLVKPGGSLKLSCAASAFTFSSYAMSWVRQTPEKRLEWVATISDGDSYTYYPD NVKGRFTISRDNAKNNLYLQMSHLKSEDTAMYYCTRDGASWDGTWFAYWGQGTLVTVSSGA KTTAPSVYPLAPVCGDTTGSSVTLGCLVKGYFPEPVTLTWNSGSLSSGVHTFPAVLQSDLYTL SSSVTVTSSTWPSQSITCNVAHPASSTKVDKKIEPRGPTIKPCPPCKCPAPNLLGGPSVFIFPPK IKDVLMISLSPIVTCVVVDVSEDDPDVQISWFVNNVEVHTAQTQTHREDYNSTLRVVSALPIQHQ DWMSGKEFKCKVNNKDLPAPIERTISKPKGSVRAPQVYVLPPPEEEMTKKQVTLTCMVTDFMP EDIYVEWTNNGKTELNYKNTEPVLDSDGSYFMYSKLRVEKKNWVERNSYSCSVVHEGLHNHH TTKSFSRTPGKGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPG VGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVP GVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGV PGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVG VPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGV GVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPG VGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGVPGVGYSEQ ID NO: 110Trastuzumab HC EVQLVESGGGLVQPGGSLRLSCAASGFNIKDTYIHWVRQAPGKGLEWVARIYPTNGYTRYADS VKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCSRWGGDGFYAMDYWGQGTLVTVSSASTK GPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSS VVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPK DTLMISRTPEVTCVWDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQ DWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYP SDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHY TQKSLSLSPGKSEQ ID NO: 111Trastuzumab LCDocket No. 028193-0052-WO01DIQMTQSPSSLSASVGDRVTITCRASQDVNTAVAWYQQKPGKAPKLLIYSASFLYSGVPSRFS GSRSGTDFTLTISSLQPEDFATYYCQQHYTTPPTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKS GTASWCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKH KVYACEVTHQGLSSPVTKSFNRGECGSEQ ID NO: 112Dulaglutide HGEGTFTSDVSSYLEEQAAKEFIAWLVKGGGGGGGSGGGGSGGGGSAESKYGPPCPPCPAP EAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREE QFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEE MTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQE G N VFSCSVM H EALH N H YTQKSLSLSLSEQ ID NO: 113Mouse Ab LC DIVMTQSPSSFSVSLGDRVTITCKASEDIYNRLAWYQQKPGNAPRLLIFGATSLETGVPSRFSG SGSGKDYTLSITSLATEDVATYYCQQYWTTPWTFGGGTKLEIKRADAAPTVSIFPPSSEQLTSG GASVVCFLNNFYPKDINVKWKIDGSERQNGVLNSWTDQDSKDSTYSMSSTLTLTKDEYERHN SYTCEATHKTSTSPIVKSFNRNECSEQ ID NO: 114Mouse Ab HC EVKLVESGGDLVKPGGSLKLSCAASAFTFSSYAMSWVRQTPEKRLEWVATISDGDSYTYYPD NVKGRFTISRDNAKNNLYLQMSHLKSEDTAMYYCTRDGASWDGTWFAYWGQGTLVTVSSGA KTTAPSVYPLAPVCGDTTGSSVTLGCLVKGYFPEPVTLTWNSGSLSSGVHTFPAVLQSDLYTL SSSVTVTSSTWPSQSITCNVAHPASSTKVDKKIEPRGPTIKPCPPCKCPAPNLLGGPSVFIFPPK IKDVLMISLSPIVTCVVVDVSEDDPDVQISWFVNNVEVHTAQTQTHREDYNSTLRVVSALPIQHQ DWMSGKEFKCKVNNKDLPAPIERTISKPKGSVRAPQVYVLPPPEEEMTKKQVTLTCMVTDFMP EDIYVEWTNNGKTELNYKNTEPVLDSDGSYFMYSKLRVEKKNWVERNSYSCSVVHEGLHNHH TTKSFSRTPGKSEQ ID NO: 115ELP Amino Acid Sequence where X is lie and Ser at a ratio of 1:1 and m is 80[VPGIG]4O-[VPGSG]4O

Claims

Docket No. 028193-0052-WO01CLAIMSWhat is claimed is:

1. A fusion protein comprising:a biologic; andan elastin-like polypeptide (ELP) attached to the biologic, wherein the ELP comprises an amino acid sequence of [VPGXG]m(SEQ ID NO: 1), wherein the ELP has a hydropathicity score of from about 0.5 to about 1.5 and m is an integer from 10 to 120,wherein the fusion protein is expressed and secreted from a eukaryotic cell.

2. The fusion protein of claim 1, wherein the ELP is attached to the C-terminus of the biologic.

3. The fusion protein of claim 1, wherein one ELP is attached to the biologic.

4. The fusion protein of claim 1, wherein more than one ELP is attached to the biologic.

5. The fusion protein of claim 1, wherein the biologic is an antibody or a Fc-fusion protein.

6. The fusion protein of claim 5, wherein the antibody is a monoclonal antibody, an immunoglobulin, or a bispecific antibody.

7. The fusion protein of claim 5, wherein the ELP is attached to a terminus of a heavy chain of the antibody or a terminus of a light chain of the antibody.

8. The fusion protein of claim 5, wherein the antibody comprises more than one ELP, each ELP being independently attached to a terminus of a heavy chain of the antibody, a terminus of a light chain of the antibody, or a combination thereof.

9. The fusion protein of claim 8, wherein:the antibody comprises two ELPs, a first ELP attached to a terminus of a first heavy chain and a second ELP attached to a terminus of a second heavy chain;Docket No. 028193-0052-WO01the antibody comprises two ELPs, a first ELP attached to a terminus of a first light chain and a second ELP attached to a terminus of a second light chain; orthe antibody comprises four ELPs, a first ELP attached to a terminus of a first heavy chain, a second ELP attached to a terminus of a second heavy chain, a third ELP attached to a terminus of a first light chain, and a fourth ELP attached to a terminus of a second light chain.

10. The fusion protein of claim 5, wherein:the Fc-fusion protein comprises one ELP attached to a terminus of a first heavy chain or a second heavy chain; orthe Fc-fusion protein comprises two ELPs, a first ELP attached to a terminus of a first heavy chain and a second ELP attached to a terminus of a second heavy chain.

11. The fusion protein of claim 5, wherein the Fc-fusion protein comprises a modified human IgG Fc domain.

12. The fusion protein of claim 11, wherein the Fc-fusion protein comprises a p35 domain, a p40 domain, an interleukin-12 (IL-12) domain, or a combination thereof.

13. The fusion protein of claim 1, wherein the ELP comprises an amino acid sequence of [VPGXG]m(SEQ ID NO: 1);wherein X is alanine (Ala), valine (Val), Serine (Ser), Isoleucine (lie), or a mixture thereof; andwherein m is an integer from 20 to 120.

14. The fusion protein of claim 13, wherein X is a mixture of Ala and Val in a ratio from 10:1 to 1:10 (Ala: Val).

15. The fusion protein of claim 13, wherein X is a mixture of Ala and Val in a ratio from 4:1 to 1:4 (Ala: Val).

16. The fusion protein of claim 13, wherein:m is 40 and X is Val or a mixture of Ala and Val in a ratio from 1:1 to 1:4 (Ala: Val); or m is 80 and X is Ala or a mixture of Ala and Val in a ratio from 1:1 to 4:1 (Ala: Val).Docket No. 028193-0052-WO0117. The fusion protein of claim 13, wherein X comprises Ser and lie arranged in blocks, each block comprising about 1 to about 40 repeats of a single residue, and wherein the overall ratio of Ser to Ile residues is from about 10:1 to about 1:10.

18. The fusion protein of claim 1, wherein the ELP comprises an amino acid sequence of [VPGVG]4o (SEQ ID NO: 2);[VPGAG]4o (SEQ ID NO: 3);[VPG[AI: VI]G]40(SEQ ID NO: 4);[VPG[A1:V4]G]40(SEQ ID NO: 5);[VPG[A4:V1]G]40(SEQ ID NO: 6);[VPGAG]8o (SEQ ID NO: 11);[VPGVG]80(SEQ ID NO: 12);[VPG[A1:V1]G]80(SEQ ID NO: 14);[VPG[A4: VI]G]80(SEQ ID NO: 15);[VPG[AI: V4]G]80(SEQ ID NO: 16);[VPGVG]120(SEQ ID NO: 21);[VPGSG]2O-[VPGIG]2O-[VPGSG]2O-[VPGIG]2O (SEQ ID NO: 17);[VPGSG]2O-[VPGIG]2O-[VPGSG]2O-[VPGIG]2O-[VPGIG]2O (SEQ ID NO: 24);[VPGSG]80(SEQ ID NO: 13); or [VPGIG]4O-[VPGSG]4O (SEQ ID NO: 115).

19. The fusion protein of claim 1, wherein the fusion protein has a glycosylation pattern characteristic of the eukaryotic cell.

20. The fusion protein of claim 1, wherein the ELP has a hydrophobicity effective to promote expression and secretion from the cell without causing precipitation or aggregation.

21. The fusion protein of claim 1, wherein the ELP has a transition temperature (Tt) that is at least 2°C higher than a temperature that the at least one eukaryotic cell is cultured at.

22. The fusion protein of claim 1, wherein the ELP has a transition temperature (Tt) of from about 34°C to about 65°C.Docket No. 028193-0052-WO0123. The fusion protein of claim 1, wherein the ELP is capable of undergoing a reversible phase transition.

24. The fusion protein of claim 1, comprising a secondary protein domain attached to the C-terminus of the ELP, wherein the secondary protein domain comprises mCherry,maltose-binding protein (MBP), thioredoxin, N-utilization substance A (NusA), green fluorescent protein (GFP), glutathione S-transferase (GST), an engineered small ubiquitin-like modifier (SUMO) tag, a modified bacterial dehalogenase tag, human serum albumin (HSA), or a second Fc-fusion protein or domain thereof.

25. The fusion protein of claim 1, wherein the fusion protein comprises a secretion domain attached to the N-terminus of the fusion protein.

26. The fusion protein of claim 25, wherein the secretion domain comprises an azurocidin signal peptide (SEQ ID NO: 28), an interleukin-2 (IL-2) signal peptide (SEQ ID NO: 31), an albumin signal peptide (SEQ ID NO: 32), a CD33 signal peptide (SEQ ID NO: 33), a tissue plasminogen activator (tPA) signal peptide (SEQ ID NO: 34), a human trypsinogen-2 signal peptide (SEQ ID NO: 35), a human chymotrypsinogen signal peptide (SEQ ID NO: 36), a maltose-binding protein (MBP) signal peptide (SEQ ID NO: 37), a FcTag, a glutathione S-transferase (GST) Tag (SEQ ID NO: 40), a CD5 signal peptide (SEQ ID NO: 38), an immunoglobulin Kappa light chain signal peptide (SEQ ID NO: 39), a trypsinogen signal peptide (SEQ ID NO: 29), or a preprolactin signal peptide (SEQ ID NO: 30).

27. A method of producing a biologic-based fusion protein, comprising:(a) engineering a eukaryotic cell to express a fusion protein comprising a biologic and an elastin-like polypeptide (ELP) attached to a C-terminus of the biologic, wherein the ELP comprises an amino acid sequence of [VPGXG]m(SEQ ID NO: 1), wherein the ELP has a hydropathicity score of from about 0.5 to about 1.5 and m is an integer from 10 to 120;(b) culturing the eukaryotic cell under conditions permitting secretion of the fusion protein into a culture supernatant; and(c) recovering the fusion protein from the culture supernatant.

28. The method of claim 27, wherein the eukaryotic cell is a mammalian cell or an insect cell.Docket No. 028193-0052-WO0129. The method of claim 28, wherein the mammalian cell is a human cell, a mouse cell, a hamster cell, a rat cell, a goat cell, or a rabbit cell.

30. The method of claim 27, wherein the fusion protein is secreted without intracellular aggregation.

31. The method of claim 27, wherein the biologic is an antibody or an Fc-fusion protein.

32. The method of claim 27, wherein the ELP comprises an amino acid sequence of [VPGXG]m(SEQ ID NO: 1);wherein X is alanine (Ala), valine (Val), Serine (Ser), Isoleucine (lie), or a mixture thereof; andwherein m is an integer from 20 to 120.

33. The method of claim 32, wherein X is a mixture of Ala and Val in a ratio from 10:1 to 1:10 (Ala: Val).

34. The method of claim 32, wherein X is a mixture of Ala and Val in a ratio from 4:1 to 1:4 (Ala: Val).

35. The method of claim 32, wherein:m is 40 and X is Val or a mixture of Ala and Val in a ratio from 1:1 to 1:4 (Ala: Val); or m is 80 and X is Ala or a mixture of Ala and Val in a ratio from 1:1 to 4:1 (Ala: Val).

36. The method of claim 32, wherein X comprises Ser and lie arranged in blocks, each block comprising about 1 to about 40 repeats of a single residue, and wherein the overall ratio of Ser to Ile residues is from about 10:1 to about 1:10.

37. The method of claim 27, wherein the ELP has a hydrophobicity effective to promote expression and secretion from the cell without causing precipitation or aggregation.

38. The method of claim 27, wherein the ELP has a transition temperature (Tt) that is at least 2°C higher than a temperature that the at least one eukaryotic cell is cultured at.Docket No. 028193-0052-WO0139. A method of purifying a fusion protein, comprising:(a) providing a culture supernatant comprising the fusion protein of claim 1;(b) inducing a phase transition of the ELP to form an insoluble phase;(c) isolating the insoluble phase from the supernatant; and(d) resolubilizing the fusion protein.

40. The method of claim 39, wherein the phase transition is induced by increasing temperature of the supernatant above a transition temperature (Tt) of the ELP.

41. The method of claim 39, wherein the phase transition is induced by adding from about 0.05M to about 5M salt to the supernatant.

42. The method of claim 41, wherein the salt is sodium chloride, ammonium sulfate, sodium citrate, sodium sulfate, magnesium sulfate, potassium sulfate, sodium phosphates, lithium chloride, sodium acetate, or sodium carbonate.

43. The method of claim 39, wherein step (c) comprises centrifugation or tangential flow filtration.

44. The method of claim 39, wherein step (d) comprises adding a low salt buffer to the fusion protein and removing insoluble contaminants using centrifugation or tangential flow filtration.

45. The method of claim 44, wherein the low salt buffer is phosphate-buffered saline (PBS), Dulbecco’s Phosphate- Buffered Saline (DPBS), citrate buffer, 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (HEPES) buffer, tris(hydroxymethyl)aminomethane (Tris) buffer, acetate buffer, histidine buffer, or succinate buffer.

46. The method of claim 39, wherein the method is performed without chromatography.

47. A method of treating a disease or disorder, comprising administering to a subject a therapeutically effective amount of the fusion protein of claim 1.Docket No. 028193-0052-WO0148. The method of claim 47, wherein the fusion protein exhibits extended serum half-life relative to the biologic without the ELP.

49. The method of claim 47, wherein the fusion protein is administered subcutaneously, intramuscularly, or intratumorally; and forms a depot at the administration site.

50. The method of claim 47, wherein the disease or disorder is cancer, an autoimmune disease, or a metabolic disorder.

51. A kit, comprising:(a) a fusion protein according to claim 1; and(b) instructions for administering or purifying the fusion protein.

52. The kit of claim 51, further comprising a buffer suitable for inducing or reversing a phase transition of the ELP, or suitable for in vivo administration.

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