Amorphous biologic suspensions and methods of use thereof

Abstract

The invention relates to a biological suspension which comprises a biologic active ingredient, a buffer, and PEG, wherein the suspension is amorphous. The invention further relates to a method for producing an amorphous biological suspension comprising mixing an aqueous buffered solution comprising a biologic and a buffer, forming a mixture, and precipitating the mixture with PEG at an incubation temperature to form an amorphous suspension.

Description

AMORPHOUS BIOLOGIC SUSPENSIONS AND METHODS OF USE THEREOFCROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims the benefit of U.S. Provisional Patent Application No.63 / 741,252, filed January 2, 2025, the entire contents of which is incorporated by reference herein.REFERENCE TO SEQUENCE LISTING SUBMITTED ELECTRONICALLY

[0002] The contents of the electronic sequence listing (26059-WO-PCT SL.xmh Size: 67.663 bytes, created on February 11, 2025) are herein incorporated by reference in their entirety.FIELD

[0003] This disclosure relates generally to a biological suspension that comprises a biologic active ingredient, a buffer, and PEG, wherein the suspension is amorphous. The invention further relates to a method for producing an amorphous biological suspension comprising mixing an aqueous buffered solution comprising a biologic and a buffer, forming a mixture, and precipitating the mixture with PEG at an incubation temperature to form an amorphous suspension.BACKGROUND

[0004] Therapeutic antibodies may be used to block cytokine activity. Subcutaneous administration is one preferred method of administration of many such antibodies, at least in part because it enables self-administration. Therapeutic antibodies are traditionally prepared in lyophilized form or in solution. Lyophilized forms may exhibit enhanced long-term stability, but require reconstitution prior to use, making them less than ideal for self-administration. Solution formulations do not require reconstitution but may suffer from reduced stability’ and typically require cold storage prior to use.

[0005] Amorphous monoclonal antibody suspensions lack long-range order crystalline structure. However, amorphous monoclonal antibodies ty pically have a degree of local order or short-range structure that arises from intermolecular interactions such as hydrogen bonding, electrostatic interactions, and van der Waals forces between the individual antibody molecules. These interactions can cause the antibody molecules to aggregate and form clusters or domains within the suspension, leading to the development of a heterogenous structure that is not crystalline but still has a certain level of order.SUMMARY

[0006] The present disclosure provides an amorphous biologic suspension, wherein the suspension comprises a biologic active ingredient. In another aspect, the invention relates to an amorphous biologic suspension, wherein the suspension comprises an antibody, antibody drug conjugate, or fusion protein. In another aspect, the invention relates to an amorphous suspension co-formulation, wherein the suspension comprises one or more biologic active ingredients. In another aspect, the invention relates to suspensions of cry stalline forms of biologic active ingredients, e.g., as amorphous slurries. In yet another aspect, the invention relates to pharmaceutical formulations comprising suspensions of amorphous forms of biologic active ingredients. In various embodiments, the amorphous form of the biologic active ingredient is used to facilitate purification, storage, and therapeutic administration of the biologies.

[0007] In an aspect, a biologic suspension is provided comprising: a) about 1 mg / ml to about 400 mg / ml of a first biologic active ingredient, wherein the first biologic active ingredient is an anti-PDl antibody; b) about 5 mM to about 100 mM buffer; and c) about 5% to about 30% polyethylene glycol (PEG); wherein the suspension has a pH between 5.0 and 9.0 and is amorphous.

[0008] In an aspect, any of the amorphous biologic suspensions described herein are provided by a method comprising:a) mixing (i) an aqueous buffered solution comprising an anti-PDl antibody, and (ii) about 5 mM to about 100 mM buffer, wherein the buffer is selected from HEPES, a Tris buffer, a phosphate buffer, MES, and MOPS; to form a mixture; andb) precipitating the mixture with PEG at an incubation temperature of about 2°C to about 40°C to form an amorphous suspension;wherein the biologic suspension has a pH above 6; and wherein the precipitation is performed by vapor diffusion, dialysis, or batch mode.

[0009] In an aspect, a method of producing a biologic amorphous suspension is provided comprising:a) mixing: (i) an aqueous buffered solution comprising a first biologic active ingredient wherein the first biologic active ingredient is an anti-PDl antibody, and (ii) about 5 mM to about 100 mM buffer, wherein the buffer is selected from HEPES, a Tris buffer, a phosphate buffer, MES, and MOPS; to form a mixture; andb) precipitating the mixture with PEG at an incubation temperature of about 2°C to about 40°C to form an amorphous suspension;wherein the suspension has a pH between 5 and 9; andwherein the precipitation is performed by vapor diffusion, dialysis, or batch mode.

[0010] In an aspect, a biologic suspension is provided comprising: (a) about 5 mg / ml to about 400 mg / ml of pembrolizumab; (b) about 5 mM to about 100 mM buffer; (c) about 5% to about 30% polyethylene glycol (PEG): (d) sodium chloride; and (e) sucrose;wherein the suspension is amorphous; and wherein the suspension is made by a method comprising:(a) mixing (i) an aqueous buffered solution comprising about 200 mg of pembrolizumab in histidine; and (ii) about 5 mM to about 100 mM buffer, wherein the buffer is selected from HEPES, a Tris buffer, a phosphate buffer. MES, and MOPS; to form a mixture; and(b) precipitating the mixture with PEG at an incubation temperature of about 2°C to about 40°C to form an amorphous suspension;wherein precipitation is by vapor diffusion, dialysis, or batch mode; andwherein the suspension has a pH between 5.0 and 9.0.

[0011] In an aspect, a biologic suspension is provided comprising: (a) about 5 mg / ml to about 400 mg / ml of pembrolizumab; (b) about 5 mM to about 100 mM buffer; (c) about 5% to about 30% polyethylene glycol (PEG); (d) sodium chloride; and (e) sucrose;wherein the suspension is amorphous; and wherein the suspension is made by a method comprising: (c) mixing (i) an aqueous buffered solution comprising about 200 mg of pembrolizumab in histidine; and (ii) about 5 mM to about 100 mM buffer, wherein the buffer is selected from HEPES, a Tris buffer, a phosphate buffer, MES, and MOPS; to form a mixture; and (d) precipitating the mixture with PEG at an incubation temperature of about 2°C to about 40°C to form an amorphous suspension;wherein precipitation is by vapor diffusion, dialysis, or batch mode; andwherein the suspension has a pH between 5.0 and 9.0; andwherein the biologic suspension additionally comprises about 260 mg of vibostolimab.

[0012] In an aspect, a method of generating a biologic amorphous suspension, the method comprising:(a) mixing together a first solution with a second solution to create a third solution, wherein (i) the first solution comprises an anti-PDl antibody and histidine, wherein the concentration of the anti-PDl antibody in the first solution is between 45 mg / ml and 55 mg / ml, and wherein the concentration of histidine in the first solution is between 18 mM and 22 mM, and wherein the first solution has a pH between 5.5 and 6.5; and (ii) the second solution comprises a buffer and PEG, wherein the concentration of the buffer in the second solution isbetween 45 mM and 55 mM, and wherein the concentration of PEG in the second solution is between 25% and 35%;(b) centrifuging the third solution to create a supernatant and a pellet, wherein the supernatant is the biologic amorphous suspension; and(c) separating the biologic amorphous suspension from the pelletBRIEF DESCRIPTION OF THE DRAWINGS

[0013] FIG. 1 shows vial images of lyo cake and upon dissolution and precipitation. The lyo cake was mixed with 20 mM Histidine, pH 5.4. The solution is then precipitated with 10 mM Hepes, pH 7.5, 30% PEG 3350.

[0014] FIG. 2 shows SONICC imaging of resulting amorphous suspension from example 2.

[0015] FIG. 3 shows vial images of lyo cake and upon precipitation (right panel). The lyo cake was precipitated with 35 mM Hepes, pH 7.5, 28 mM arginine, 15% PEG3350.

[0016] FIG. 4 shows a visible photomicrograph of anti-PD-1 (197mg / ml), 33 mM Hepes, pH 8.0, 19% PEG 3350 of Example 11.

[0017] FIG. 5 shows a visible photomicrograph of an anti-TIGIT formulation comprising 180 mg / ml anti-TIGIT antibody, 33 mM Hepes, pH 8.0, 19% PEG 3350 of Example 12.

[0018] FIG. 6 shows a visible photomicrograph of an anti-TIGIT formulation comprising 194 mg / ml anti-TIGIT antibody, 33 mM Hepes, pH 8.0, 19% PEG 3350, 33 mM NaCl, 33 mM arginine of Example 13. Furthermore, this FIG. shows the benefits of an amorphous suspension comprising arginine (over FIG. 5) with respect to morphology leading to improved syringeability.

[0019] FIG. 7 shows a visible photomicrograph of an anti-PD-1 and anti-TIGIT combination (192 mg / ml, 33 mM Hepes, pH 8.0, 19% PEG 3350, 33 mM NaCl, 33 mM arginine of example 14.

[0020] FIG. 8A shows a full spectrum of13C cross polarization (CP) magic angle spinning (MAS) spectra of aTIGIT (vibostolimab) amorphous suspension.

[0021] FIG. 8B shows an enlarged spectral region of13C cross polarization (CP) magic angle spinning (MAS) spectra of aTIGIT (vibostolimab) amorphous suspension.

[0022] FIG. 9 shows a split image Micro-ED image of amorphous anti-PD-1 suspension on the left and crystalline anti-PD-1 on the right of Example 22.

[0023] FIG. 10 SONICC imaging of resulting amorphous suspension from example 23.

[0024] FIG. 11 A shows full spectrum of13C (carbon-13) cross polarization (CP) magic angle spinning (MAS) spectra of pembrolizumab crystalline suspension, amorphous suspension, liquid high concentration formulation, and lyophilized formulation (from top to bottom).

[0025] FIG. 1 IB shows an enlarged spectral region of13C (carbon-13) cross polarization (CP) magic angle spinning (MAS) spectra of pembrolizumab crystalline suspension, amorphous suspension, liquid high concentration formulation, and lyophilized formulation (from top to bottom).

[0026] FIG 12 shows SONICC characterization by visible UV and SHG as described in Example 27 A.

[0027] FIG 13 shows SONICC characterization by visible UV and SHG as described in Example 27B.

[0028] FIG. 14A shows a photomicrograph of amorphous pembrolizumab before rheology.

[0029] FIG. 14B shows a photomicrograph of amorphous pembrolizumab after rheology.

[0030] FIG. 15A provides data showing steady-state viscosity as a function of shear rate, for a pembrolizumab suspension sample.

[0031] FIG. 15B provides data showing the equilibrium total strain plotted as a function of shear rate, for a pembrolizumab suspension sample.

[0032] FIG. 16A shows ID13C spectra of an amorphous suspension of pembrolizumab at 200 mg / ml, collected before (upper panel) and after (lower panel) rheology.

[0033] FIG. 16B shows an overlay of the13C spectra of the upper and lower panels from FIG.16 A.DETAILED DESCRIPTION

[0034] The invention provides a biologic suspension comprising: a) about 150 mg / ml to about 400 mg / ml of a biologic active ingredient; b) about 5mM to about 100 mM buffer; and c) about 5% to about 30% polyethylene glycol (PEG); wherein the suspension has a pH between 6.0 and 9.0 and is amorphous.

[0035] In some embodiments, the invention provides a biologic suspension comprising: a) about 150 mg / ml to about 400 mg / ml of a biologic active ingredient comprising an anti-PDl antibody: b) about 5mM to about 100 mM buffer; and c) about 5% to about 30% polyethylene glycol (PEG); wherein the suspension has a pH between 6.0 and 9.0 and is amorphous. In some embodiments, the anti-PDl antibody is an anti-human PD1 antibody. In some embodiments, the anti-PDl antibody is pembrolizumab.

[0036] In some embodiments, a biologic suspension is provided comprising: a) about 150 mg / ml to about 400 mg / ml of a biologic active ingredient comprising an antibody drug conjugate (ADC); b) about 5mM to about 100 mM buffer; and c) about 5% to about 30% polyethylene glycol (PEG); wherein the suspension has a pH between 6.0 and 9.0 and is amorphous. In some embodiments, the ADC is produced by recombinant DNA technology. In some embodiments, the ADC is a ROR1 ADC. In some embodiments, the ADC is an aTau ADC.

[0037] In some embodiments, a biologic suspension is provided comprising: a) about 150 mg / ml to about 400 mg / ml of a biologic active ingredient comprising a T-cell engager; b) about 5mM to about 100 mM buffer; and c) about 5% to about 30% polyethylene glycol (PEG); wherein the suspension has a pH between 6.0 and 9.0 and is amorphous.

[0038] In some embodiments, a biologic suspension is provided comprising: a) about 150 mg / ml to about 400 mg / ml of a biologic active ingredient comprising an anti-TIGIT antibody; b) about 5mM to about 100 mM buffer; and c) about 5% to about 30% polyethylene glycol (PEG); wherein the suspension has a pH between 6.0 and 9.0 and is amorphous. In some embodiments, the anti-TIGIT antibody is vibostolimab.

[0039] In some embodiments, a biologic suspension is provided comprising: a) about 150 mg / ml to about 400 mg / ml of a biologic active ingredient comprising an anti-RSV antibody; b) about 5mM to about 100 mM buffer; and c) about 5% to about 30% polyethylene glycol (PEG); wherein the suspension has a pH between 6.0 and 9.0 and is amorphous. In some embodiments, the monoclonal antibody is an anti-RSV antibody. In some embodiments, the anti-RSV antibody is clesrovimab.

[0040] In some embodiments of the invention, the biologic suspension comprises a high concentration of a biologic active ingredient. In some embodiments, the biologic active ingredient is an anti-PDl antibody. In some embodiments, the anti-PDl antibody is pembrolizumab. In some embodiments, the biologic active ingredient is an ADC. In some embodiments, the ADC is a ROR1 ADC. In some embodiments, the biologic active ingredient is a T-cell engager.

[0041] In some embodiments of the invention, the PEG is PEG 3350. In specific embodiments of the invention, the concentration of PEG 3350 is 10 mg / ml. In specific embodiments of the invention, the molecular weight of the PEG is from about 2,500 to about 20,000. In specific embodiments of the invention, the biologic suspension comprises 0-30% PEG 3350. In specific embodiments of the invention, the biologic suspension comprises 0-20% PEG 3350. In specific embodiments of the invention, the biologic suspension comprises 5-15% PEG 3350. In specific embodiments of the invention, the precipitant solution comprises 0-20% PEG 3350 or any othersuitable value. In specific embodiments of the invention, the PEG is present in an amount of 15 % weight / volume (w / v). In specific embodiments, the PEG is present in an amount of 25 % weight / volume (w / v).

[0042] In some embodiments of the invention, the buffer is HEPES, a Tris buffer, a phosphate buffer, MES, or MOPS. In some embodiments of the invention, the buffer is 5-100 mM. In specific embodiments of the invention, the concentration of the buffer is 5-75 mM. In specific embodiments of the invention, the concentration of the buffer is 10-50 mM. In specific embodiments of the invention, the concentration of the buffer is 10 mM. In yet another embodiment, the concentration of the buffer is 50 mM.

[0043] In specific embodiments of the invention, the buffer is HEPES. In specific embodiments of the invention, the HEPES buffer is present at a concentration of about 5-100 mM. In specific embodiments of the invention, the HEPES buffer is present at a concentration of about 5-75 mM. In specific embodiments of the invention, the HEPES buffer is present at a concentration of about 10-50 mM. In specific embodiments of the invention, the HEPES buffer is present at a concentration of about 10 mM. In specific embodiments of the invention, the HEPES buffer is present at a concentration of about 50 mM.

[0044] In some embodiments of the invention, the biologic suspension comprises histidine. In specific embodiments of the invention, the histidine buffer is present at a concentration of about 20 mM. In specific embodiments of the invention, the histidine buffer is present at a concentration of about 10 mM.

[0045] In some embodiments of the invention, the biologic suspension comprises arginine. In specific embodiments of the invention, the arginine concentration is about 20-50 mM. In specific embodiments of the invention, the arginine concentration is about 50 mM. In specific embodiments of the invention, the arginine benefits morphology of the suspension. In specific embodiments of the invention, the arginine enables individual small particles within the suspension. In a specific embodiment of the invention, a suspension comprising arginine improves syringeability of the suspension.

[0046] In some embodiments of the invention, the biologic suspension comprises about 6% to about 8% weight / volume (w / v) sucrose, trehalose or (2- hydroxypropyl)-P-cyclodextrin. In specific embodiments of the invention, the suspension comprises about 6% to about 8% weight / volume (w / v) sucrose. In specific embodiments, the suspension comprises sodium chloride. In specific embodiments of the invention, the sodium chloride concentration is 150 mM.

[0047] In some embodiments of the invention, the suspension has a pH between 7 and 8.5. In some embodiments of the invention, the suspension has a pH between 6.2 and 8.2. In specific embodiments of the invention, the suspension has a pH of 7.

[0048] The invention provides a biologic suspension comprising: a) about 150 mg / ml to about 400 mg / ml of a biologic active ingredient comprising an antibody drug conjugate (ADC); b) about 5mM to about 100 mM buffer; and c) about 5% to about 30% polyethylene glycol (PEG); wherein the suspension has a pH between 6.0 and 9.0 and is amorphous.

[0049] In some embodiments of the invention, the biologic active ingredient is an antibody drug conjugate (ADC). In specific embodiments of the invention, the antibody within the ADC is produced by recombinant DNA technology. In specific embodiments of the invention, the ADC is ROR1-ADC. In specific embodiments, the ADC is an anti-Tau ADC.

[0050] The invention provides a biologic suspension comprising: a) about 150 mg / ml to about 400 mg / ml of a biologic active ingredient comprising an T-cell engager; b) about 5mM to about 100 mM buffer; and c) about 5% to about 30% polyethylene glycol (PEG); wherein the suspension has a pH between 6.0 and 9.0 and is amorphous.

[0051] In specific embodiments of the invention, the biologic is a T-cell engager.

[0052] In some embodiments of the invention, the biologic active ingredient is an antibody or antigen binding fragment thereof. In specific embodiments of the invention, the biologic is an antibody. In specific embodiments of the invention, the antibody is produced by recombinant DNA technology. In specific embodiments of the invention, the antibody is a monoclonal antibody. In specific embodiments of the invention, the monoclonal antibody is an IgGl or IgG4 antibody. In specific embodiments of the invention, the monoclonal antibody is anti-PD-1 antibody.

[0053] In specific embodiments of the invention, the anti-PD-1 antibody is an anti -human PD-1 antibody. In specific embodiments of the invention, the anti -human PD-1 antibody or antigen binding fragment thereof comprises three light chain CDRs comprising CDRL 1 of SEQ ID NO: 1, CDRL2 of SEQ ID NO:2 and CDRL3 of SEQ ID NO:3 and three heavy chain CDRs of CDRH1 of SEQ ID NO:6, CDRH2 of SEQ ID NO:7 and CDRH3 of SEQ ID NO:8. In specific embodiments of the invention, the anti-PD-1 antibody is pembrolizumab. In specific embodiments of the invention, the present disclosure provides a method for producing high concentration drug substance of pembrolizumab.

[0054] In specific embodiments of the invention, the monoclonal antibody is an anti-TIGIT antibody. In specific embodiments of the invention, the anti-TIGIT antibody is vibostolimab.

[0055] In specific embodiments of the invention, the monoclonal antibody is an anti-RSV antibody. In specific embodiments of the invention, the monoclonal antibody is clesrovimab.

[0056] In some embodiments of the invention, the biologic suspension further comprises an additional biologic active ingredient. In specific embodiments of the invention, the biologic suspension comprises an anti-TIGIT antibody and an anti-PD-1 antibody. In specific embodiments of the invention, the biologic suspension comprises vibostolimab and pembrolizumab. In specific embodiments of the invention, the biologic suspension comprises 260 mg of vibostolimab and 260 mg of pembrolizumab. In specific embodiments of the invention, the biologic suspension comprises a vibostolimab and pembrolizumab in a single injection.

[0057] In some embodiments of the invention, the biologic suspension is produced by a method comprising: a) mixing i. an aqueous buffered solution comprising the biologic active ingredient, and ii. about 5 mM to about 100 mM buffer, wherein the buffer is selected from HEPES. a Tris buffer, a phosphate buffer, MES, and MOPS; to form a mixture; and b) precipitating the mixture with PEG at an incubation temperature of about 2°C to about 40°C to form a suspension; wherein the biologic suspension has a pH above 6 and is amorphous; and wherein the precipitation is performed by vapor diffusion, dialysis, or batch mode.

[0058] In some embodiments of the invention, the biologic suspension is encapsulated into a delivery system. In some embodiments of the invention, the delivery system is a patch or hydrogel. In specific embodiments of the invention, the delivery system is a hydrogel. In specific embodiments of the invention, the delivery system is a patch.

[0059] Also provided herein are methods for producing a biologic suspension comprising a) mixing i. an aqueous buffered solution comprising the biologic active ingredient, and ii. about 5 mM to about 100 mM buffer, wherein the buffer is selected from HEPES, a Tris buffer, a phosphate buffer, MES, and MOPS; to form a mixture; and b) precipitating the mixture with PEG at an incubation temperature of about 2°C to about 40°C to form an suspension; wherein the biologic suspension has a pH above 6 and is amorphous; and wherein the precipitation is performed by vapor diffusion, dialysis, or batch mode.

[0060] In specific embodiments of the invention, the biologic active ingredient is pembrolizumab and the buffer is histidine. In specific embodiments of the invention, the concentration of the biologic active ingredient is 150 mg / ml to about 400 mg / ml. In specific embodiments of the invention, the concentration of the PEG is about 5% to 30 %. In specific embodiments of the invention, the pH is between 6 and 9. In specific embodiments of the invention, the PEG is PEG 3350. In specific embodiments of the invention, the concentration ofPEG 3350 is 10 mg / ml. In specific embodiments of the invention, the molecular weight of the PEG is from about 2,500 to about 20,000. In specific embodiments of the invention, the PEG is 15 % weight / volume (w / v). In specific embodiments of the invention, the PEG is 25 % weight / volume (w / v). In specific embodiments of the invention, the buffer is HEPES. In specific embodiments of the invention, the buffer is about 5-100 mM HEPES. In specific embodiments of the invention, the buffer is about 5-75 mM HEPES. In specific embodiments of the invention, the buffer is about 10-50 mM HEPES. In specific embodiments of the invention, the buffer is about 10 mM HEPES. In specific embodiments of the invention, the buffer is about 50 mM HEPES. In specific embodiments of the invention, the suspension further comprises histidine. In specific embodiments of the invention, the histidine concentration is about 20 mM. In specific embodiments of the invention, the suspension further comprises arginine. In specific embodiments of the invention, the suspension further comprises about 20-50 mM arginine. In specific embodiments of the invention, the arginine concentration is about 50 mM. In specific embodiments of the invention, the suspension further comprises about 6% to about 8% weight / volume (w / v) sucrose, trehalose or (2- hydroxypropyl)-0-cyclodextrin. In specific embodiments of the invention, the suspension further comprises about 6% to about 8% weight / volume (w / v) sucrose. In specific embodiments of the invention, the suspension further comprises sodium chloride. In specific embodiments of the invention, the suspension further comprises about 150 mM sodium chloride. In specific embodiments of the invention, the suspension has a pH between 6.2 and 8.2. In specific embodiments of the invention, the suspension has a pH between 7 and 9. In specific embodiments of the invention, the suspension has a pH between 7 and 8.5. In specific embodiments of the invention, the suspension has a pH of 7.

[0061] Also provided herein are methods for producing a biologic suspension comprising a) mixing i. an aqueous buffered solution comprising the biologic active ingredient, wherein the biologic active ingredient is an antibody or an antibody drug conjugate (ADC), and ii. about 5 mM to about 100 mM buffer, wherein the buffer is selected from HEPES, a Tris buffer, a phosphate buffer, MES, and MOPS; to form a mixture; and b) precipitating the mixture with PEG at an incubation temperature of about 2°C to about 40°C to form an suspension; wherein the biologic suspension has a pH above 6 and is amorphous; and wherein the precipitation is performed by vapor diffusion, dialysis, or batch mode.

[0062] In some embodiments of the invention, the biologic active ingredient is an antibody drug conjugate (ADC). In specific embodiments of the invention, the biologic active ingredient is an antibody drug conjugate (ADC), wherein the antibody within the ADC is produced byrecombinant DNA technology. In specific embodiments of the invention, the ADC is R0R1-ADC. In specific embodiments of the invention, the ADC is an anti-Tau ADC.

[0063] Also provided herein are methods for producing a biologic suspension comprising a) mixing i. an aqueous buffered solution comprising the biologic active ingredient comprising an antibody or a T-cell engager, and ii. about 5 mM to about 100 mM buffer, wherein the buffer is selected from HEPES, a Tris buffer, a phosphate buffer, MES, and MOPS; to form a mixture; and b) precipitating the mixture with PEG at an incubation temperature of about 2°C to about 40°C to form an suspension; wherein the biologic suspension has a pH above 6 and is amorphous; and wherein the precipitation is performed by vapor diffusion, dialysis, or batch mode

[0064] In some embodiments of the invention, the biologic active ingredient is a T-cell engager. In specific embodiments of the invention, the biologic active ingredient is an antibody or antigen binding fragment thereof. In specific embodiments of the invention, the biologic is an antibody. In specific embodiments of the invention, the antibody is produced by recombinant DNA technology.

[0065] In specific embodiments of the invention, the antibody is a monoclonal antibody. In specific embodiments of the invention, the monoclonal antibody is an IgGl or IgG4 antibody. In specific embodiments of the invention, the monoclonal antibody is anti-PD-1 antibody. In specific embodiments of the invention, the anti-PD-1 antibody is an anti-human PD-1 antibody. In specific embodiments of the invention, the anti -human PD-1 antibody or antigen binding fragment thereof comprises three light chain CDRs comprising CDRL1 of SEQ ID NO: 1, CDRL2 of SEQ ID NO:2 and CDRL3 of SEQ ID NO:3 and three heavy chain CDRs of CDRH1 of SEQ ID NO:6, CDRH2 of SEQ ID NO:7 and CDRH3 of SEQ ID NO:8. In specific embodiments of the invention, the anti-PD-1 antibody is pembrolizumab.

[0066] In specific embodiments of the invention, the monoclonal antibody is an anti-TIGIT antibody. In specific embodiments of the invention, the anti-TIGIT antibody is vibostolimab. In specific embodiments of the invention, the monoclonal antibody is clesrovimab.

[0067] In some embodiments of the invention, the biologic suspension further comprises an additional biologic active ingredient. In specific embodiments of the invention, the biologic suspension comprises an anti-TIGIT antibody and an anti-PD-1 antibody. In specific embodiments of the invention, the biologic suspension comprises vibostolimab and pembrolizumab. In specific embodiments of the invention, the biologic suspension comprises 260 mg of an anti-TIGIT antibody and 260 mg of pembrolizumab. In specific embodiments of26059the invention, the biologic suspension comprises an anti-TIGIT antibody and pembrolizumab in a single injection device.

[0068] In some embodiments of the invention, the biologic suspension is encapsulated into a delivery system. In some embodiments of the invention, the delivery system is a patch or hydrogel. In specific embodiments of the invention, the delivery system is a hydrogel. In specific embodiments of the invention, the delivery system is a patch.

[0069] Also provided herein is an biologic suspension comprising: a) about 5 mg / ml to about 400 mg / ml pembrolizumab; b) about 5 mM to about 100 mM buffer; c) about 5% to about 30% polyethylene glycol (PEG); d) sodium chloride; e) sucrose; wherein the suspension is amorphous; and wherein the suspension is made by a method comprising: a) mixing an aqueous buffered solution comprising pembrolizumab in histidine; and about 5 mM to about 100 mM buffer, wherein the buffer is selected from HEPES, a Tris buffer, a phosphate buffer, MES, and MOPS; to form a mixture; and b) precipitating the mixture with PEG at an incubation temperature of about 2°C to about 40°C to form an suspension which is amorphous; wherein precipitation is performed by vapor diffusion, dialysis, or batch mode; and wherein the suspension has a pH between 6.0 and 9.0.

[0070] Also provided herein is an biologic suspension comprising: a) about 5 mg / ml to about 400 mg / ml of a biologic active ingredient; b) about 5 mM to about 100 mM buffer; c) about 5% to about 30% polyethylene glycol (PEG); d) sodium chloride; e) sucrose; wherein the suspension is made by a method for producing an anti-PD-1 antibody suspension comprising: a) mixing i. an aqueous buffered solution comprising about 200 mg of pembrolizumab in histidine; ii. about 5 mM to about 100 mM buffer, wherein the buffer is selected from HEPES, a Tris buffer, a phosphate buffer, MES, and MOPS; to form a mixture; and b) precipitating the mixture with PEG at an incubation temperature of about 2°C to about 40°C to form an suspension, wherein precipitation is performed by vapor diffusion, dialysis, or batch mode; and wherein the suspension has a pH between 6.0 and 9.0 and is amorphous; and wherein the biologic suspension additional comprises about 260 mg of vibostolimab.

[0071] In some embodiments of the invention, the suspension was recovered by centrifugation.

[0072] In some embodiments, the anti-PDl antibody is an anti-human PD1 antibody. In some embodiments, the anti-human PD-1 is encapsulated in a hydrogel microsphere. In specific embodiments, pembrolizumab is encapsulated in a hydrogel microsphere.

[0073] In some embodiments, the method comprises an incubation temperature of 2-40°C. In some embodiments, the method comprises agitation. In some embodiments, the method comprises temperature ramping from 4°C to 10-40°C.26059

[0074] In some embodiments of the invention, the present disclosure provides characterization of the amorphous and re-dissolved amorphous suspensions using biochemical and biophysical methods (e g., SONICC, UPSEC, C13 Solid-State NMR, Micro-ED and injectability).

[0075] In some embodiments, the monoclonal antibody suspensions of the invention have an average particle size between 0.5 and 20 microns. In a further aspect, the resulting amorphous suspensions had a particle size of 0.5-50 microns. In some embodiments, the monoclonal suspensions comprise an anti-PDl antibody.

[0076] In one aspect, the disclosure provides a method to produce high concentration amorphous drug substance, wherein the drug substance is shown to have acceptable viscosity properties for administration.

[0077] Also provided herein are the attributes of the re-dissolved amorphous biologic (ex. monoclonal antibodies) are consistent with the intact biologic starting sample in biochemical and biophysical characterization studies.

[0078] In some embodiments, these methods are amenable to multiple pharmaceutical applications such as purification (manufacture), storage, formulation, and drug delivery.

[0079] In some embodiments, the conditions provided herein provide a scalable one step amorphous suspension process as a platform approach which can be formulated as a high concentration, low volume, formulation in a pre-filled syringe with room temperature stability under sterile conditions.

[0080] In some embodiments, the monoclonal antibody suspensions of the invention are obtainable by lyo cake solids and batch precipitation methods. In one embodiment, the monoclonal antibody suspensions of the invention are obtainable by lyo cake solids.

[0081] In other embodiments, the invention relates to suspensions of the amorphous antibodies of the invention, including those at higher concentrations and lower viscosities than would be possible with a corresponding non-amorphous solution at the same concentration of antibody. In some embodiments the suspension of the antibody has an antibody concentration of greater than about 200 mg / ml. In some embodiments the suspension of the antibody has an antibody concentration of greater than about 260 mg / ml. In other embodiments, the antibody suspensions of the invention have increased stability, i.e., they maintain biological activity7of the antibody longer than corresponding solution formulations. In some embodiments, the increased stability is at room temperature, enabling storage of the suspensions of the invention at room temperature rather than at 4°C (weeks to months). In some embodiments, the antibody comprises an anti-PDl antibody.

[0082] In one aspect, the conditions provide a suspension that is stable at room temperature.26059

[0083] In some embodiments, the suspension of the present invention is delivered subcutaneously. In yet other embodiments, the suspension of the invention is delivered, e.g., by inhalation or insufflation.

[0084] In another aspect, the invention relates to methods of preparing a biologic suspension of the invention. In one embodiment, the biologic suspension is made by a batch precipitation method comprising the steps of mixing a solution of the antibody, antibody drug conjugate, or fusion proteins with a precipitant solution to form an amorphous suspension, incubating that precipitating solution for a time sufficient for amorphous formation, and harvesting the amorphous suspension from the solution. In some embodiments, an equal volume of precipitant solution is added to form the amorphous suspension.

[0085] In another aspect, the invention relates to methods of preparing an amorphous biologic suspension of the invention from a lyo cake. Suspensions made upon precipitation from lyo cake solids allows for high concentrations > 200 mg / ml (in some embodiments. > 260 mg / ml) underreconstitution and is the most viable method for large scale batch preparation of amorphous suspensions for manufacturing applications into prefilled syringes suitable for subcutaneous drug delivery.

[0086] In an aspect, a method of generating a biologic amorphous suspension is provided, the method comprising:(a) mixing together a first solution with a second solution to create a third solution, wherein (i) the first solution comprises an anti-PDl antibody and histidine, wherein the concentration of the anti-PDl antibody in the first solution is between 45 mg / ml and 55 mg / ml, and wherein the concentration of histidine in the first solution is between 18 mM and 22 mM, and wherein the first solution has a pH between 5.5 and 6.5; and (ii.) the second solution comprises a buffer and PEG, wherein the concentration of the buffer in the second solution is between 45 mM and 55 mM, and wherein the concentration of PEG in the second solution is between 25% and 35%;(b) centrifuging the third solution to create a supernatant and a pellet, wherein the supernatant is the biologic amorphous suspension; and(c) separating the biologic amorphous suspension from the pellet.

[0087] In some embodiments, the concentration of the anti-PDl antibody in the first solution is about 50 mg / ml.

[0088] In some embodiments, the anti -human PD-1 antibody comprises three light chain CDRs comprising a CDRL1 comprising the amino acid sequence of SEQ ID NO: 1, a CDRL2 comprising the amino acid sequence of SEQ ID NO: 2 and a CDRL3 comprising the amino acid26059sequence of SEQ ID NO:3; and three heavy chain CDRs comprising a CDRH1 comprising the amino acid sequence of SEQ ID NO:6, a CDRH2 comprising the amino acid sequence of SEQ ID NO:7 and a CDRH3 comprising the amino acid sequence of SEQ ID NO: 8.

[0089] In some embodiments, the anti-PD-1 antibody comprises pembrolizumab.

[0090] In some embodiments, the concentration of histidine in the first solution is about 20 mM.

[0091] In some embodiments, the buffer in the second solution is Hepes buffer.

[0092] In some embodiments, the concentration of PEG in the second solution is about 30%.

[0093] In some embodiments, PEG is PEG 3350.

[0094] The summary of the technology described above is non-limiting and other features and advantages of the technology will be apparent from the following detailed description, and from the claims.PD-1 antagonists, anti-human PD-1 monoclonal antibodies (anti-PD-1, aPD-1 or aPDl) useful in the invention

[0095] In some embodiments, the monoclonal antibody is an anti-PD-1 antibody. In a further embodiment, the anti-PD-1 antibody is pembrolizumab.

[0096] Examples of mAbs that bind to human PD-1, useful in the suspensions, treatment methods, compositions, and uses of the invention, are described in US 7,521 ,051 , US 8,008,449, and US 8,354,509. Specific anti-human PD-1 mAbs useful as the PD-1 antagonist in the suspensions, treatment methods, compositions, and uses of the invention include: pembrolizumab (formerly known as MK-3475, SCH 900475 and lambrolizumab), a humanized IgG4 mAb with the structure described in WHO Drug Information, Vol. 27, No. 2, pages 161-162 (2013) and which comprises the heavy and light chain amino acid sequences shown in Figure 7, and the humanized antibodies h409All, h409A16 and h409A17, which are described in WO 2008 / 156712.

[0097] Provided herein are PD-1 antagonists or anti-human PD-1 monoclonal antibodies that can be used in any of the suspensions, methods, compositions, kits, and uses disclosed herein, including any chemical compound or biological molecule that blocks binding of PD-L1 to PD-1 and preferably also blocks binding of PD-L2 to PD-1.

[0098] Any monoclonal antibodies that bind to a PD-1 polypeptide, a PD-1 polypeptide fragment, a PD-1 peptide, or a PD-1 epitope and block the interaction between PD-1 and its ligand PD-L1 or PD-L2 can be used. In some embodiments, the anti-human PD-1 monoclonal antibody binds to a PD-1 polypeptide, a PD-1 polypeptide fragment, a PD-1 peptide, or a PD-126059epitope and blocks the interaction between PD-1 and PD-L1. In other embodiments, the antihuman PD-1 monoclonal antibody binds to a PD-1 polypeptide, a PD-1 polypeptide fragment, a PD-1 peptide, or a PD-1 epitope and blocks the interaction between PD-1 and PD-L2. In yet other embodiments, the anti -human PD-1 monoclonal antibody binds to a PD-1 polypeptide, a PD-1 polypeptide fragment, a PD-1 peptide, or a PD-1 epitope and blocks the interaction between PD-1 and PD-L1 and the interaction between PD-1 and PD-L2.

[0099] Any monoclonal antibodies that bind to a PD-L1 polypeptide, a PD-L1 polypeptide fragment, a PD-L1 peptide, or a PD-L1 epitope and block the interaction between PD-L1 and PD-1 can also be used.

[0100] In certain embodiments, the anti -human PD-1 monoclonal antibody is selected from the group consisting of pembrolizumab, nivolumab, cemiplimab, dostarlimab, pidilizumab (U.S. Pat. No. 7,332,582), AMP-514 (Medlmmune LLC, Gaithersburg, MD), PDR001 (U.S. Pat. No. 9,683,048), BGB-A317 (U.S. Pat. No. 8,735,553), and MGA012 (MacroGenics, Rockville, MD). In one embodiment, the anti-human PD-1 monoclonal antibody is pembrolizumab. In another embodiment, the anti -human PD-1 monoclonal antibody is nivolumab. In another embodiment, the anti-human PD-1 monoclonal antibody is cemiplimab. In one embodiment, the anti-human PD-1 monoclonal antibody is dostarlimab. In yet another embodiment, the anti-human PD-1 monoclonal antibody is pidilizumab. In one embodiment, the anti -human PD-1 monoclonal antibody is AMP-514. In another embodiment, the anti-human PD-1 monoclonal antibody is PDR001. In yet another embodiment, the anti -human PD-1 monoclonal antibody is BGB-A317. In still another embodiment, the anti-human PD-1 monoclonal antibody is MGA012.

[0101] In some embodiments, an anti-human PD-1 antibody or antigen binding fragment thereof for use in the methods and uses of the invention comprises three light chain CDRs of CDRL1, CDRL2 and CDRL3 and / or three heavy chain CDRs of CDRH1, CDRH2 and CDRH3.

[0102] In one embodiment of the invention, CDRL1 has the amino acid sequence as set forth in SEQ ID NO: 1 or a variant of the amino acid sequence as set forth in SEQ ID NO: 1, CDRL2 has the amino acid sequence as set forth in SEQ ID NO: 2 or a variant of the amino acid sequence as set forth in SEQ ID NO:2, and CDRL3 has the amino acid sequence as set forth in SEQ ID NO:3 or a variant of the amino acid sequence as set forth in SEQ ID NO:3.

[0103] In one embodiment, CDRH1 has the amino acid sequence as set forth in SEQ ID NO:6 or a variant of the amino acid sequence as set forth in SEQ ID NO:6, CDRH2 has the amino acid sequence as set forth in SEQ ID NO:7 or a variant of the amino acid sequence as set forth in SEQ ID NO:7, and CDRH3 has the amino acid sequence as set forth in SEQ ID NO: 8 or a variant of the amino acid sequence as set forth in SEQ ID NO: 8.26059

[0104] In one embodiment, the anti-human PD-1 antibody comprises three light chain CDRs comprising a CDRL1 comprising the amino acid sequence of SEQ ID NO: 1, a CDRL2 comprising the amino acid sequence of SEQ ID NO:2 and a CDRL3 comprising the amino acid sequence of SEQ ID NO: 3 and three heavy chain CDRs comprising a CDRH1 comprising the amino acid sequence of SEQ ID NO: 6, a CDRH2 comprising the amino acid sequence of SEQ ID NO:7 and a CDRH3 comprising the amino acid sequence of SEQ ID NO: 8.

[0105] In an alternative embodiment of the invention, the anti -human PD-1 antibody comprises three light chain CDRs comprising a CDRL1 comprising the amino acid sequence of SEQ ID NO: 11. a CDRL2 comprising the amino acid sequence of SEQ ID NO: 12 and a CDRL3 comprising the amino acid sequence of SEQ ID NO: 13 and three heavy chain CDRs comprising a CDRH1 comprising the amino acid sequence of SEQ ID NO: 16, a CDRH2 comprising the amino acid sequence of SEQ ID NO: 17 and a CDRH3 comprising the amino acid sequence of SEQ ID NO: 18.

[0106] In one embodiment, the three-light chain CDRs have the amino acid sequences as set forth in SEQ ID NO:1, SEQ ID NO:2, and SEQ ID NO:3 and the three heavy chain CDRs have the amino acid sequences as set forth in SEQ ID NO:6, SEQ ID NO:7 and SEQ ID NO:8.

[0107] In an alternative embodiment, the three-light chain CDRs have the amino acid sequences as set forth in SEQ ID NO: 11, SEQ ID NO: 12. and SEQ ID NO: 13 and the three heavy chain CDRs have the amino acid sequences as set forth in SEQ ID NO: 16, SEQ ID NO: 17 and SEQ ID NO: 18.

[0108] Some anti-human PD-1 antibody and antigen binding fragments of the invention comprise a light chain variable region and a heavy’ chain variable region. In some embodiments, the light chain variable region comprises the amino acid sequence as set forth in SEQ ID NO:4 or a variant of the amino acid sequence as set forth in SEQ ID NO:4, and the heavy chain variable region comprises the amino acid sequence as set forth in SEQ ID NO: 9 or a variant of the amino acid sequence as set forth in SEQ ID NO:9. In further embodiments, the light chain variable region comprises the amino acid sequence as set forth in SEQ ID NO: 14 or a variant of the amino acid sequence as set forth in SEQ ID NO: 14, and the heavy chain variable region comprises the amino acid sequence as set forth in SEQ ID NO: 19 or a variant of the amino acid sequence as set forth in SEQ ID NO:19In such embodiments, a light chain variable region or heavy' chain variable region sequence is identical to the reference sequence except having one, two. three, four or five amino acid substitutions. In some embodiments, the substitutions are in the framework region (i.e., outside of the CDRs). In some embodiments, one, two, three, four or five of the amino acid substitutions are conservative substitutions.26059

[0109] In one embodiment of the methods, kits or uses of the invention, the anti-human PD-1 antibody or antigen binding fragment comprises a light chain variable region comprising or consisting of the amino acid sequence as set forth in SEQ ID NO:4 and a heavy chain variable region comprising or consisting of the amino acid sequence as set forth in SEQ ID NO: 9. In a further embodiment, the anti-human PD-1 antibody or antigen binding fragment comprises a light chain variable region comprising or consisting of the amino acid sequence as set forth in SEQ ID NO: 14 and a heavy chain variable region comprising or consisting of the amino acid sequence as set forth in SEQ ID NO: 19.

[0110] In another embodiment, the methods, kits or uses of the invention comprise an antihuman PD-1 antibody or antigen binding protein that has a VL domain and / or a VH domain with at least 99%, 98%, 97%, 96%, 95%, 90%, 85%, 80%, 75% or 50% sequence homology to one of the VL domains or VH domains described above and exhibits specific binding to PD-1. In another embodiment, the anti -human PD-1 antibody or antigen binding protein of the methods the invention comprises VL and VH domains having up to 1, 2, 3, 4, or 5 or more amino acid substitutions, and exhibits specific binding to PD-1.[OHl] In any of the embodiments above, the PD-1 antagonist may be a full-length anti-PD-1 antibody or an antigen binding fragment thereof that specifically binds human PD-1. In certain embodiments, the PD-1 antagonist is a full-length anti-PD-1 antibody selected from any class of immunoglobulins, including IgM, IgG, IgD, IgA, and IgE. Preferably, the antibody is an IgG antibody. Any isot pe of IgG can be used, including IgGi, IgGz. IgG?. and IgG4. Different constant domains may be appended to the VL and VH regions provided herein. For example, if a particular intended use of an antibody (or fragment) of the invention were to call for altered effector functions, a heavy chain constant domain other than IgGl may be used. Although IgGl antibodies provide for long half-life and for effector functions, such as complement activation and antibody -dependent cellular cytotoxicity, such activities may not be desirable for all uses of the antibody. In such instances an IgG4 constant domain, for example, may be used.

[0112] In embodiments of the invention, the PD-1 antagonist is an anti-PD-1 antibody comprising a light chain comprising or consisting of a sequence of amino acid residues as set forth in SEQ ID NO:5 and a heavy chain comprising or consisting of a sequence of amino acid residues as set forth in SEQ ID NO: 10. In alternative embodiments, the PD-1 antagonist is an anti-PD-1 antibody comprising a light chain comprising or consisting of a sequence of amino acid residues as set forth in SEQ ID NO: 15 and a heavy chain comprising or consisting of a sequence of amino acid residues as set forth in SEQ ID NO:20. In some methods of the26059invention, the PD-1 antagonist is pembrolizumab or a pembrolizumab biosimilar. In some methods of the invention, the PD-1 antagonist is nivolumab or a nivolumab biosimilar.

[0113] Ordinarily, amino acid sequence variants of the anti-PD-1 antibodies and antigen binding fragments of the invention will have an amino acid sequence having at least 75% amino acid sequence identity with the amino acid sequence of a reference antibody or antigen binding fragment e.g.(e.g., heavy chain, light chain, VH, VL, or humanized sequence), or at least 80%, at least 85%, at least 90%, or at least 95%, 98%, or 99%. Identity or homology7with respect to a sequence is defined herein as the percentage of amino acid residues in the candidate sequence that are identical with the anti-PD-1 residues, after aligning the sequences and introducing gaps, if necessary, to achieve the maximum percent sequence identity7, and not considering any conservative substitutions as part of the sequence identity. None of N-terminal, C-terminal, or internal extensions, deletions, or insertions into the antibody sequence shall be construed as affecting sequence identity or homology.

[0114] Sequence identity refers to the degree to which the amino acids of two polypeptides are the same at equivalent positions when the two sequences are optimally aligned. Sequence identity can be determined using a BLAST algorithm wherein the parameters of the algorithm are selected to give the largest match between the respective sequences over the entire length of the respective reference sequences. The following references relate to BLAST algorithms often used for sequence analysis: BLAST ALGORITHMS: Altschul, S.F., et al., (1990) J. Mol. Biol.215:403-410; Gish, W., et al., (1993) Nature Genet. 3:266-272; Madden, T.L., et al., (1996) Meth. Enzymol. 266:131-141; Altschul, S.F., et al., (1997) Nucleic Acids Res. 25:3389-3402; Zhang, J., et al., (1997) Genome Res. 7:649-656; Wootton, J.C., et al., (1993) Comput. Chem.17:149-163; Hancock, J.M. et al., (1994) Comput. Appl. Biosci. 10:67-70; ALIGNMENT SCORING SYSTEMS: Dayhoff, M.O., et al., "A model of evolutionary change in proteins." in Atlas of Protein Sequence and Structure, (1978) vol. 5, suppl. 3. M.O. Dayhoff (ed.), pp. 345-352, Natl. Biomed. Res. Found., Washington, DC; Schwartz, R.M.. et al., "Matrices for detecting distant relationships." in Atlas of Protein Sequence and Structure. (1978) vol. 5, suppl. 3." M.O. Dayhoff (ed ), pp. 353-358, Natl. Biomed. Res. Found., Washington, DC: Altschul, S.F., (1991) J. Mol. Biol. 219:555-565; States, D.J., et al., (1991) Methods 3:66-70; Henikoff, S., et al., (1992) Proc. Natl. Acad. Sci. USA 89:10915-10919; Altschul, S.F.. et al., (1993) J. Mol. Evol.36:290-300; ALIGNMENT STATISTICS: Karlin, S.. et al., (1990) Proc. Natl. Acad. Sci. USA 87:2264-2268; Karlin, S„ et al., (1993) Proc. Natl. Acad. Sci. USA 90:5873-5877; Dembo, A., et al., (1994) Ann. Prob. 22:2022-2039; and Altschul, S.F. "Evaluating the statistical significance of26059multiple distinct local alignments." in Theoretical and Computational Methods in Genome Research (S. Suhai, ed.), (1997) pp. 1-14, Plenum, New York.

[0115] Likewise, either class of light chain can be used in the suspension, compositions and methods herein. Specifically, kappa, lambda, or variants thereof are useful in the suspensions, compositions and methods of the invention.Table 1. Exemplary7PD-1 Antibody Sequences26059Anti-PD-1 antibody dosing

[0116] In some embodiments, the suspension and / or biologic active ingredient is administered subcutaneously, on a weekly, biweekly, triweekly, every 4 weeks, every 5 weeks, every 6 weeks, monthly, bimonthly, or quarterly basis at about 10, about 20, about 50, about 80, about 100, about 200, about 300, about 400, about 500, about 1000 or about 2500 mg / subject.

[0117] In some specific embodiments, the dose of the suspension and / or biologic active ingredient from about 0.01 mg / kg to about 50 mg / kg, from about 0.05 mg / kg to about 25 mg / kg, from about 0.1 mg / kg to about 10 mg / kg, from about 0.2 mg / kg to about 9 mg / kg, from about 0.3 mg / kg to about 8 mg / kg, from about 0.4 mg / kg to about 7 mg / kg, from about 0.5 mg / kg to about 6 mg / kg, from about 0.6 mg / kg to about 5 mg / kg, from about 0.7 mg / kg to about 4 mg / kg, from about 0.8 mg / kg to about 3 mg / kg. from about 0.9 mg / kg to about 2 mg / kg. from about 1.0 mg / kg to about 1.5 mg / kg, from about 1.0 mg / kg to about 2.0 mg / kg, from about 1.0 mg / kg to about 3.0 mg / kg, or from about 2.0 mg / kg to about 4.0 mg / kg.

[0118] In some specific embodiments, the dose of the suspension and / or biologic active ingredient is from about 10 mg to about 500 mg, from about 25 mg to about 500 mg. from about 50 mg to about 500 mg, from about 100 mg to about 500 mg, from about 200 mg to about 500 mg, from about 150 mg to about 250 mg, from about 175 mg to about 250 mg, from about 200 mg to about 250 mg, from about 150 mg to about 240 mg, from about 175 mg to about 240 mg, or from about 200 mg to about 240 mg. In some embodiments, the dose of the suspension and / or biologic active ingredient is about 50 mg, about 75 mg, about 100 mg, about 125 mg, about 15026059mg, about 175 mg, about 200 mg, about 225 mg, about 240 mg, about 250 mg, about 300 mg, about 400 mg, or about 500 mg.

[0119] In another embodiment of the invention, the PD-1 antagonist in the suspension is pembrolizumab, or a pembrolizumab variant, which is administered in a liquid medicament at a dose selected from the group consisting of 1 mg / kg Q2W, 2 mg / kg Q2W, 3 mg / kg Q2W, 5 mg / kg Q2W, 10 mg / kg Q2W, 1 mg / kg Q3W, 2 mg / kg Q3W, 3 mg / kg Q3W, 5 mg / kg Q3W, or 10 mg / kg Q3W. In other embodiments, the PD-1 antagonist in the suspension is pembrolizumab, or a pembrolizumab variant, which is administered in a liquid medicament at a flat dose such as 200 mg Q3W or 400 mg Q6W.

[0120] In some embodiments of the suspensions, methods, compositions, kits and uses described herein, the anti -human PD-1 antibody (e.g., anti-PD-1 monoclonal antibody) or antigen binding fragment thereof is pembrolizumab, and a human patient is administered about 200 mg, about 240 mg, about 400 mg, about 480 mg, or about 2 mg / kg pembrolizumab once every three or six weeks.

[0121] In one embodiment, a human patient is administered about 200 mg pembrolizumab once every' three weeks. In one embodiment, a human patient is administered about 240 mg pembrolizumab once every three weeks. In one embodiment, a human patient is administered 2 mg / kg pembrolizumab once every three weeks. In one embodiment, the human patient is administered 400 mg pembrolizumab once every three weeks.

[0122] In certain embodiments of the suspensions, methods, compositions, kits and uses described herein, the anti -human PD-1 monoclonal antibody or antigen binding fragment thereof is pembrolizumab and the human patient is administered 400 mg pembrolizumab once every six weeks.

[0123] In some embodiments of the suspensions, methods, compositions, kits and uses described herein, the anti -human PD-1 monoclonal antibody or antigen binding fragment thereof is pembrolizumab, and the human patient is administered about 200 mg, about 240 mg, about 400 mg, about 480 mg, or about 2 mg / kg pembrolizumab once every six weeks. In one embodiment, the human patient is administered about 200 mg pembrolizumab once every six weeks. In one embodiment, the human patient is administered about 240 mg pembrolizumab once every' six weeks. In one embodiment, the human patient is administered about 400 mg pembrolizumab once every six weeks. In one embodiment, the human patient is administered 480 mg pembrolizumab once every' six weeks. In one embodiment, the human patient is administered 2 mg / kg pembrolizumab once every' six weeks.26059

[0124] In certain embodiments of the suspension, methods, compositions, kits and uses described herein, the anti -human PD-1 monoclonal antibody or antigen binding fragment thereof is pembrolizumab, and the human patient is administered about 200 mg pembrolizumab once every three weeks. In certain embodiments of the suspensions, methods, compositions, kits and uses described herein, the anti-human PD-1 monoclonal antibody or antigen binding fragment thereof is pembrolizumab, and the human patient is administered about 400 mg pembrolizumab once every7six weeks.

[0125] In other embodiments, the amorphous biologic suspension is administered by subcutaneous injection.

[0126] In other embodiments, the selected dose of pembrolizumab is administered by subcutaneous injection.

[0127] In some embodiments, the selected dose of pembrolizumab is administered subcutaneously. In one embodiment, pembrolizumab is administered by subcutaneous injection at a dose of about 260 mg to about 450 mg. In embodiments of the invention, the amount of pembrolizumab administered subcutaneously to the patient is from 260 mg to 420 mg, 320 mg to 420 mg, from 340 mg to 420 mg, from 345 mg to 415 mg, from 350 mg to 410 mg, from 355 mg to 405 mg, from 360 mg to 400 mg, from 365 mg to 395 mg, from 370 mg to 390 mg, from 375 mg to 385 mg, or from 379 mg to 381 mg. In a further embodiment of the invention, pembrolizumab is administered by subcutaneous injection at a dose of about 260 mg to about 450 mg. In yet a further embodiment of the invention, pembrolizumab is administered subcutaneously at a dose of about 320 mg to about 450 mg. In yet a further embodiment of the invention, pembrolizumab is administered subcutaneously at a dose of about 260 mg.

[0128] In embodiments of the invention, pembrolizumab is administered subcutaneously to the patient, wherein the pembrolizumab is part of a suspension and is present in the amorphous biologic suspension at a concentration of 130 mg / mL. In embodiments of the invention, pembrolizumab administered subcutaneously to the patient, wherein the pembrolizumab is part of the amorphous biologic suspension and is present in the amorphous biologic suspension at a concentration of 165 mg / mL. In embodiments of the invention, pembrolizumab is administered subcutaneously to the patient in two injections. In embodiments of the invention, the amount of pembrolizumab administered subcutaneously to the patient is 260 mg in one pre-filled syringe. In embodiments of the invention, the amount of pembrolizumab administered subcutaneously to the patient is 380 mg in one pre-filled syringe. In embodiments of the invention, the amount of pembrolizumab administered subcutaneously to the patient is 260 mg in two pre-filled syringes.26059In embodiments of the invention, the amount of pembrolizumab administered subcutaneously to the patient is 380 mg in two pre-filled syringes.TIGIT antagonists, anti- TIGIT monoclonal antibodies useful in the invention

[0129] In some embodiments, the monoclonal antibody is an anti-TIGIT antibody. In some embodiments, the monoclonal antibody is vibostolimab. In specific embodiments of the invention, the anti-TIGIT antibody comprises a light and heavy chain sequence of SEQ ID Nos 21 and 22. In specific embodiments of the invention, the anti-TIGIT antibody comprises three light chain CDRs comprising CDRL1 of SEQ ID NO: 23. CDRL2 of SEQ ID NO: 24, CDRL3 of SEQ ID NO: 25, CDRH1 of SEQ ID NO: 26, CDRH2 of SEQ ID NO: 27, CDRH3 of SEQ ID NO: 28. In specific embodiments of the invention, the anti-TIGIT antibody comprises a heavy chain variable region of SEQ ID NO: 29 and a light chain variable region of SEQ ID NO: 30.Table 2. Exemplary TIGIT Antibody Sequences26059aRSV - anti-RSV antibodies useful in the invention

[0130] In some embodiments, the monoclonal antibody is an anti-RSV antibody. In some embodiments, the anti-RSV antibody is clesrovimab. In some embodiments, the anti-RSV antibody is disclosed in WO2017075124A1Table 3. Exemplary anti-RSV Antibody Sequences26059

[0131] In specific embodiments of the invention, the anti-RSV antibody comprises a heavy¬ chain variable region of SEQ ID NO: 37 and a light chain variable region of SEQ ID NO: 38.

[0132] In specific embodiments of the invention, the anti-RSV antibody comprises three light chain CDRs comprising CDRL1 of SEQ ID NO: 31, CDRL2 of SEQ ID NO: 32, CDRL3 of SEQ ID NO: 33, CDRH1 of SEQ ID NO: 34, CDRH2 of SEQ ID NO: 35, CDRH3 of SEQ ID NO: 36. In specific embodiments of the invention, the anti-RSV antibody is a full-length antibody¬ having two light chains and two heavy chains, wherein each light chain comprises: a variable region comprising SEQ ID NO: 38 and a human kappa light chain (SEQ ID NO: 44); and each heavy chain comprises: a variable region comprising SEQ ID NO: 37 and a human IgGl constant region (SEQ ID NO: 43).

[0133] In specific embodiments of the invention, the anti-RSV antibody comprises a heavychain comprising the amino acid sequence of SEQ ID NO: 53 and a light chain comprising the amino acid sequence of SEQ ID NO: 55.

[0134] In specific embodiments of the invention, the anti-RSV antibody- comprises the heavy chain consists of the amino acid sequence of SEQ ID NO: 53 and the amino acid sequence of the light chain consists of the ammo acid sequence of SEQ ID NO: 55.Anti-Tau antibody useful in the inventionThe invention comprises the anti-Tau antibody- as disclosed in WO2021262791.

[0135] In specific embodiments of the invention, the anti-Tau antibody comprises a light and heavy chain sequence of SEQ ID Nos 57 and 58.

[0136] In specific embodiments of the invention, the anti-Tau antibody comprises three light chain CDRs comprising CDRL1 of SEQ ID NO: 59, CDRL2 of SEQ ID NO: 60, CDRL3 of SEQ26059ID NO: 61, CDRH1 of SEQ ID NO: 62, CDRH2 of SEQ ID NO: 63, CDRH3 ofSEQ ID NO: 64.

[0137] In specific embodiments of the invention, the anti-Tau antibody comprises a heavy chain variable region of SEQ ID NO: 66 and a light chain variable region of SEQ ID NO: 65.ROR1 ADC useful in the invention

[0138] The invention comprises an amorphous suspension comprising the ROR1-ADC as disclosed in PCT / US18 / 039112, a buffer, and PEG, as described herein.Definitions

[0139] Listed below are definitions of various terms used herein. These definitions apply to the terms as they are used throughout this specification and claims, unless otherwise limited in specific instances, either individually or as part of a larger group.26059

[0140] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art. Generally, the nomenclature used herein and the laboratory procedures in cell culture, molecular genetics, organic chemistry, and peptide chemistry are those well-known and commonly employed in the art.

[0141] As used herein, the articles “a” and “an” refer to one or to more than one (i. e. , to at least one) of the grammatical object of the article. By way of example, “an element” means one element or more than one element. Furthermore, use of the term “including” as well as other forms, such as “include,” “includes,” and “included,” is not limiting.

[0142] As used herein, the term “about” in quantitative terms refers to plus or minus 10% of the value it modifies (rounded up to the nearest whole number if the value is not sub-dividable, such as a number of molecules or nucleotides).

[0143] All ranges disclosed herein are inclusive of the recited endpoint and independently combinable (for example, the range of “from 50 mg to 500 mg” is inclusive of the endpoints, 50 mg and 500 mg, and all the intermediate values). The endpoints of the ranges and any values disclosed herein are not limited to the precise range or value; they are sufficiently imprecise to include values approximating these ranges and / or values.

[0144] As used herein, the term “comprising” may include the embodiments “consisting of’ and “consisting essentially of.” The terms “comprise(s),” “include(s).” “having,” “has.” “may.” “contain(s),” and variants thereof, as used herein, are intended to be open-ended transitional phrases, terms, or words that require the presence of the named ingredients / steps and permit the presence of other ingredients / steps. However, such description should be construed as also describing compositions or processes as “consisting of’ and “consisting essentially of’ the enumerated components, which allows the presence of only the named components or compounds, along with any acceptable carriers or fluids, and excludes other components or compounds.

[0145] As used herein, including the appended claims, the singular forms of words such as “a,” “an,” and “the,” include their corresponding plural references unless the context clearly dictates otherwise. All references cited herein are incorporated by reference to the same extent as if each individual publication, patent application, or patent, w as specifically and individually indicated to be incorporated by reference. Citation of the references herein is not intended as an admission that any of the foregoing is pertinent prior art. nor does it constitute any admission as to the contents or date of these publications or documents.

[0146] “Administration” and “treatment,” as it applies to an animal, human, experimental subject, cell, tissue, organ, or biological fluid, refers to contact of an exogenous pharmaceutical,26059therapeutic, diagnostic agent, or composition to the animal, human, subject, cell, tissue, organ, or biological fluid. “Administration” and “treatment” can refer, e.g., to therapeutic, pharmacokinetic, diagnostic, research, and experimental methods. Treatment of a cell encompasses contact of a reagent to the cell, as well as contact of a reagent to a fluid, where the fluid is in contact with the cell. “Administration” and “treatment” also include in vitro and ex vivo treatments, e.g., of a cell, by a reagent, diagnostic, binding composition, or by another cell.

[0147] As used herein, the term "hypervariable region" refers to the amino acid residues of an antibody that are responsible for antigen-binding. The hypervariable region comprises amino acid residues from a "complementarity determining region" or "CDR" e.g.(e.g., residues 24-34 (CDRL1), 50-56 (CDRL2) and 89-97 (CDRL3) in the light chain variable domain and residues 31-35 (CDRH1), 50-65 (CDRH2) and 95-102 (CDRH3) in the heavy chain variable domain (Kabat et al. (1991) Sequences of Proteins of Immunological Interest, 5th Ed. Public Health Service, National Institutes of Health, Bethesda, Md.) and / or those residues from a "hypervariable loop" (i.e. , residues 26-32 (LI), 50-52 (L2) and 91-96 (L3) in the light chain variable domain and 26-32 (Hl), 53-55 (H2) and 96-101 (H3) in the heavy chain variable domain (Chothia and Lesk (1987) J. Mol. Biol. 196: 901-917). As used herein, the term "framework" or "FR" residues refers to those variable domain residues other than the hypervariable region residues defined herein as CDR residues. The residue numbering above relates to the Kabat numbering system and does not necessarily correspond in detail to the sequence numbering in the accompanying Sequence Listing.

[0148] “Conservatively modified variants” or “conservative substitution” refers to substitutions of amino acids are known to those of skill in this art and may be made without altering the biological activity of the resulting molecule, even in essential regions of the polypeptide. Such exemplary substitutions may be made in accordance with those set forth in Table 4 as follows:Table 4: Exemplary Conservative Amino Acid Substitutions26059

[0149] In addition, those of skill in this art recognize that, in general, single amino acid substitutions in non-essential regions of a polypeptide do not alter biological activity. See, e.g., Watson et al. (1987) Molecular Biology of the Gene, The Benjamin / Cummings Pub. Co., p. 224 (4th Edition).

[0150] The phrase "consists essentially of," or variations such as "consist essentially of' or "consisting essentially of," as used throughout the specification and claims, indicate the inclusion of any recited elements or group of elements, and the optional inclusion of other elements, of similar or different nature than the recited elements, that do not materially change the basic or novel properties of the specified dosage regimen, method, or composition. As a non-limiting example, a binding compound that consists of a recited amino acid sequence may also include one or more amino acids, including substitutions of one or more amino acid residues that do not materially affect the properties of the binding compound.

[0151] ‘‘Biologic active ingredient” includes monoclonal antibodies, proteins (e.g., fusion proteins), nucleic acid molecules, and antibody drug conjugates.

[0152] “Immune condition” or “immune disorder” encompasses, e.g.. pathological inflammation, an inflammatory disorder, and an autoimmune disorder or disease. “Immune condition” also refers to infections, persistent infections, and proliferative conditions, such as cancer, tumors, and angiogenesis, including infections, tumors, and cancers that resist eradication by the immune system. “Cancerous condition” includes, e.g., cancer, cancer cells, tumors, angiogenesis, and precancerous conditions such as dysplasia.

[0153] “Inflammatory disorder” means a disorder or pathological condition where the pathology' results, in whole or in part, from, e.g., a change in number, change in rate of migration, or change in activation, of cells of the immune system. Cells of the immune system include, e.g., T cells, B cells, monocytes or macrophages, antigen presenting cells (APCs), dendritic cells, microglia, NK cells, NKT cells, neutrophils, eosinophils, mast cells, or any other cell specifically associated with the immunology7, for example, cytokine-producing endothelial or epithelial cells.26059

[0154] As used herein, “inhibit” or “treat” or “treatment” includes a postponement of development of the symptoms associated with autoimmune disease or pathogen-induced immunopathology and / or a reduction in the severity of such symptoms that will or are expected to develop. The terms further include ameliorating existing uncontrolled or unwanted autoimmune-related or pathogen-induced immunopathology symptoms, preventing additional symptoms, and ameliorating or preventing the underlying causes of such symptoms. Thus, the terms encompass, but are not limited to, circumstances in which a beneficial result has been conferred on a vertebrate subject with an autoimmune or pathogen-induced immunopathology disease or symptom, or with the potential to develop such a disease or symptom.

[0155] To examine the extent of inhibition of PD-1 activity, for example, samples or assays comprising a given, e.g., protein, gene, cell, or organism, are treated with a potential inhibiting agent, such as an anti PD-1 mAb,

[0156] “Specifically.” or “selectively” binds, when referring to a ligand / receptor. antibody / antigen, or other binding pair, indicates a binding reaction which is determinative of the presence of the protein in a heterogeneous population of proteins and other biologies. Thus, under designated conditions, a specified ligand binds to a particular receptor and does not bind in a significant amount to other proteins present in the sample. As used herein, an antibody is said to bind specifically to a polypeptide comprising a given sequence (in this case PD-1) if it binds to polypeptides comprising the sequence of aPD-1 but does not bind to proteins lacking the sequence of aPD-1. For example, an antibody that specifically binds to a polypeptide comprising aPD-1 may bind to a FLAG® -tagged form of PD-1 but will not bind to other FLAG®-tagged proteins.

[0157] “MES” is 2-(N-morpholino)ethanesulfonic acid.

[0158] “MOPS” is 3-morpholinopropane-l -sulfonic acid.

[0159] “HEPES” is (4-(2-Hydroxyethyl)piperazine-l -ethane-sulfonic acid).

[0160] “Tris” is Tris(hydroxymethyl)aminomethane.

[0161] When used with reference to an amorphous antibody suspension of the present invention, "concentration" refers to the amount of antibody present in each macroscopic unit volume of solution. The term concentration is used in its customary' sense despite the inherent heterogeneity' of the suspension, as compared to a traditional solution. The concentration of antibody in an amorphous suspension is equal to the concentration of an equivalent sample in which the antibody was not in amorphous form.

[0162] The present invention provides amorphous forms of antibodies, fusion proteins, and antibody drug conjugates. Furthermore, the present invention provides amorphous forms of26059monoclonal antibodies, suspensions of these and pharmaceutical formulations of these suspensions. Highly purified aPD-1 monoclonal antibody, anti-RSV antibody, anti-HMPV antibody, anti-TIGIT antibody , anti-RSV antibody, R0R1-ADC, and aTau antibody were used in batch screening experiments. Amorphous suspensions were obtained by lyo cake and high concentrated liquid preparations. Conditions were established to prepare amorphous suspensions by bulk crystallization methods (batch and lyo cake) in high yield. In some embodiments, the resulting amorphous suspensions have a particle size of 0.5-30 microns. In other embodiments, the resulting amorphous suspensions have a particle size of 0.5-10 microns. Amorphous biologic suspensions of the present invention have several advantageous properties for use in therapy. The amorphous suspensions of the biologies of certain embodiments of the invention can also be prepared at higher concentrations those solution formulations. This high concentration can enable more efficient administration to subject, e.g., by subcutaneous injection. For example, solution formulations at 100 mg / ml cannot be used to deliver more than 100 mg to a subject with a single subcutaneous injection due to limitations of how much volume can be practically delivered at a single injection site. This limits dosing to approximately 1.5 mg / kg unless the subject is willing to accept (and in some cases administer) multiple injections at multiple sites. The amorphous suspensions of the present invention, in contrast, can be used to prepare pharmaceutical formulations up to 250 mg / ml or more, enabling higher dosing with lower injection volume, and thus less discomfort. The reduced volume decreased viscosity and use of a smaller needle are all likely to decrease patient discomfort accompanying subcutaneous administration of therapeutic monoclonal antibodies, which is of particular concern when a drug of intended for selfadministration (e.g., by prefilled syringe or auto injector). Additionally, the higher concentration of the amorphous biologic suspension allows for administration with a single pre-filled syringe.

[0163] Amorphous suspensions can be characterized through methods known in the art. In some embodiments, solid-state NMR (SSNMR) and small-angle X-ray scattering have been used to characterize protein suspensions. In some embodiments, SSNMR has been demonstrated to assess the crystallinity of protein suspensions (Mingyue Li, Paul Reichert, Chakravarthy Narasimhan, Bradley Sorman, Wei Xu, Aaron Cote, Yongchao Su, Mol. Pharmaceutics 2022, 19, 3, 936-952) and to differentiate polymorphs in cry stalline pembrolizumab suspensions (Patrick Larpent, Lorenzo Codan, Jameson R. Bothe, Luca Iuzzolino, Suzette Pabit, Sudipta Gupta, Thierry Fischmann, Yongchao Su. Paul Reichert, Dirk Stueber. Aaron Cote, Mol. Pharmaceutics 2024, 21, 8, 4024-4037).

[0164] Although the liquid and amorphous show comparable pK and BA, the amorphous suspensions have advantage of lower viscosity at higher concentrations in the delivery options26059and possibly room temperature stability. The amorphous suspensions aPD-1 antibodies of the present invention also exhibit comparable properties about the pharmacokinetics of drug deliver}'. Compared with the corresponding solution formulations, the amorphous suspensions of the present invention exhibit comparable bioavailability.

[0165] Amorphous biologic suspensions of the present invention also have other advantageous properties. Suspensions of the amorphous biologies will have improved stability compared with corresponding solution formulations, i.e., the amorphous suspensions will retain biological activity for a longer time. Suspensions of the amorphous antibodies of the present invention can even be stored at room temperature, whereas typical solution formulations would have to be stored at 4°C. The longer shelf-life, and the ability to store the suspensions of the present invention a room temperature, offer significant advantages in handling of drug product and supply chain management.

[0166] The precipitation methods of the present invention also provide a method of storing purified biologies even if such amorphous suspensions are re-dissolved prior to use. In one embodiment, a biologic is produced and at least partially purified by methods described elsewhere herein and known in the art. The biologic active ingredient is then precipitated, e.g., by batch precipitation as described in Example 2. or by bulk dialysis. The resulting concentrated suspension could act as a stable concentrated preparation suitable for shipping and reformulating at global formulation sites. Other important advantage is to extend the platform approach for other IgGl, IgG4, bi-specific, tri-specific antibodies as well as ADCs and fusion proteins.EXAMPLES

[0167] The following examples are meant to be illustrative and should not be construed as further limiting. The contents of the figures and all references, patents, and published patent applications cited throughout this application are expressly incorporated herein by reference.Example 1: Preparation of lyo cake for amorphous pembrolizumab experiments

[0168] A mixture of 50 mg / ml pembrolizumab and 10 mM histidine, pH 5.4, (8 ml) was dialyzed in a 100 KDa cutoff dialysis bag versus 100 ml of distilled w ater and exchanged 2X over 18 hours. The resulting volume increased to about 9 ml total during dialysis. 0.256 ml of 50% PEG 3350 (W / V) solution was added to a final 1.55% PEG 3350 concentration. The resulting solution at 30 mg / ml pembrolizumab by UV A260Z 280 nm was added to 32 x 2 ml sterile vials, each fitted with a septum cap. A lyophilization recipe run in a Lyo Star 2 FTS Systems from SP Scientific Products, Warminster, PA is show n below in Table 5.26059Table 5. Parameters for preparation of lyophilized cake for amorphous pembrolizumab monoclonal antibody.Example 2: Preparation of amorphous suspension of pembrolizumab from lyo cake example 1. (Low Concentration)

[0169] 31 pl of 20 mM histidine, pH 5.4 was added to the resulting cake (8.4 mg / vial) from Example 1 via a pipette. After a reconstitution time of 2 hours at room temperature, the cake dissolved completely and had a consistency similar to that of mineral oil as shown in the middle panel of FIG. 1. For precipitation, 31 pl of 20 mM Hepes, pH 7.5, 30% PEG 3350 was subsequently added, and the vial was mixed on a MediMixlOO® automated mixer (Health mark, CA) set at the lowest speed at room temperature. The clear solution was observed to change to a white paste slowly within 5 minutes at room temperature. The final amorphous suspension was measured at 133 mg / ml by 1 / 10 dilution in PBS by UV A260Z 280 nm reading. SONICC analyses shown in FIG. 2 was run on a 1 / 0 dilution (10 mM Hepes, pH 7.5, 8 % PEG 3350) of the resulting paste confirming conversion to an amorphous suspension as shown in the right panel of FIG. 1.Example 3. Preparation of a high concentration amorphous suspension of pembrolizumab from lyo cake example 1.

[0170] 476 pl of 35 mM Hepes, pH 7.5, 28 mM arginine, 15% PEG 3350 was added to the 12 vials of lyo cake (8.4 mg / vial) from Example 1 via a pipette. After a reconstitution time of 2 hours at room temperature, the cake dissolved completely and had a consistency similar to that of a cream as shown in the right hand of FIG.3. The vial was mixed on a MediMixlOO® automated mixer (Health mark, CA) set at the lowest speed at room temperature. The lyo cake changed to a26059white paste slowly within 2 hours at room temperature. The final amorphous suspension was measured at 211 mg / ml by 1 / 10 dilution in PBS by UV A260 / 280 nm reading. SONICC analyses shown in FIG. 4 was run on a 1 / 0 dilution (10 mM Hepes, pH 7.0, 8 % PEG 3350) of the resulting paste confirming conversion to an amorphous suspension as shown in the right panel of FIG. 4Example 4. Preparation of amorphous suspension of anti-PD-1 antibody (pembrolizumab)

[0171] 666 pl of 50 mM HEPES, pH 7.0 containing 25 % w / v PEG 3350 was mixed with 333 pl of 40 mg / ml pembrolizumab in 20 mM histidine. pH 5.4 solution at RT. Precipitation was carried out in batch mode at a total volume of 1 mL, with each batch yielding -13 mg of the antibody. The precipitation mixture was kept at room temperature for 4 hours while rotating at 12 rpm on a tube mixer. An amorphous pembrolizumab suspension was recovered by centrifugation at 1000 RCF for 5 minutes. A280 UV measurements were made of the resulting pellet suspension (68 mg / ml) and supernatant (0.1 mg / ml). SONICC inspection by Visible, UV+ and SHG-imaging confirmed a particle suspension consistent with an amorphous suspension.Example 5. Preparation of amorphous suspension of anti-RSV antibody (clesrovimab)

[0172] 666 pl of 50 mM HEPES, pH 7.0 containing 25 % w / v PEG was mixed with 333 pl of 40 mg / ml of anti-RSV antibody (clesrovimab) in 20 mM histidine, pH 5.4 solution at room temperature. Precipitation was carried out in batch mode at a total volume of 1 mL, yielding -13 mg of the antibody. Solutions were prepared with distilled water and were filtered with a 0.22-micron filter. The precipitation mixture was kept at room temperature for 4 hours while rotating at 12 rpm on a tube mixer. An amorphous pembrolizumab suspension was recovered by centrifugation at 1000 RCF for 5 minutes. A280 UV measurements were made of the resulting pellet suspension (62 mg / ml) and supernatant (0.1 mg / ml). SONICC inspection by Visible, UV+ and SHG- imaging confirmed a particle suspension consistent with an amorphous suspension.Example 6. Attempted Preparation of amorphous suspension of anti-PD-1 antibody

[0173] 666 pl of 20 mM histidine, pH 5.4 containing 25 % w / v PEG was mixed with 333 pl of 40 mg / ml pembrolizumab in 20 mM histidine, pH 5.4 solution at room temperature. Precipitation was carried out in batch mode at total volume of 1 mL, with each batch yielding -13 mg of the antibody. Solutions were prepared with distilled water and were filtered with a 0.22-micron filter. The precipitation mixture w as kept at room temperature for 4 hours while rotating at 12 rpm on a tube mixer.26059Example 7. Preparation of amorphous solid forms of anti-RSV antibody (clesrovimab)

[0174] 666 pl of 50 mM HEPES, pH 7.0 containing 25 % w / v PEG was mixed with 333 pl of 40 mg / ml of anti-RSV antibody (clesrovimab) in 20 mM histidine, pH 5.4 solution at room temperature. Precipitation was carried out in batch mode at total volume of 1 mL, yielding ~13 mg of the antibody. Solutions were prepared with distilled water and were filtered with a 0.22-micron filter. The precipitation mixture was kept at room temperature for 4 hours while rotating at 12 rpm on a tube mixer. Amorphous aTIGIT suspension was recovered by centrifugation at 1000 RCF for 5 minutes. A280 UV measurements were made of the resulting pellet suspension (100 mg / ml) and supernatant (0.1 mg / ml). SONICC inspection by Visible, UV+ and SHG-imaging confirmed a particle suspension consistent with an amorphous suspension.Example 8. Attempted Preparation of amorphous suspension of anti-HMPV antibody

[0175] 666 pl of 20 m histidine, pH 5.4 containing 25 % w / v PEG was mixed with 333 pl of 70 mg / ml aRSV (RBI 1) in 20 mM sodium glutamate, pH 5.4 solution at RT. Solutions were prepared with distilled water and were filtered with a 0.22 -micron filter. Precipitation mixture was kept at room temperature for 4 hours while rotating at 12 rpm on a tube mixer.Example 9. Preparation of amorphous solid suspension of anti-TIGIT antibody (vibostolimab)

[0176] 666 pl of 50 mM HEPES, pH 8.0 containing 25 % w / v PEG was mixed with 333 pl of 70 mg / ml of anti-TIGIT antibody (vibostolimab) in 20 mM histidine, pH 5.4 solution at room temperature. Precipitation was carried out in batch mode at a total volume of 1 mL, yielding ~22 mg of the antibody. Solutions were prepared with distilled water and were filtered with a 0.22-micron filter. The precipitation mixture was kept at room temperature for 4 hours while rotating at 12 rpm on a tube mixer. An amorphous aTIGIT suspension was recovered by centrifugation at 1000 RCF in a Beckman Coulter Alegra X-15R centrifuge using a SX-4750 rotor for 5 minutes. A280 UV measurements were made of the resulting pellet suspension (139 mg / ml) and supernatant (0.1 mg / ml) after 1:10 dilution in PBS. SONICC inspection by Visible, UV+ and SHG-imaging confirmed a particle suspension consistent with an amorphous suspension.Example 10. Preparation of co-formulation amorphous suspension of anti-PD-1 antibody (pembrolizumab) and aTIGIT (vibostolimab), pH 8.0

[0177] 666 pl of 50 mM HEPES, pH 8.0 containing 25 % w / v PEG was mixed with 166 pl of 40 mg / ml pembrolizumab in 20 mM histidine, pH 5.4 solution and 166 pl of 70 mg / ml of anti-26059TIGIT antibody (vibostolimab) in 20 mM histidine, pH 5.4 at RT (turbid). Precipitation was carried out in batch mode at total volume of 1 mL, with each batch yielding ~13 mg of the antibody. Precipitation mixture was kept at room temperature for 4 hours while rotating at 12 rpm on a tube mixer. The amorphous coformulation suspension was recovered by centrifugation at 1000 RCF in a Beckman Coulter Alegra X-15R centrifuge using a SX-4750 rotor for 5 minutes. A280 UV measurements were made of the resulting pellet suspension (57 mg / ml) and supernatant (0.1 mg / ml). SONICC inspection by Visible, UV+ and SHG- imaging confirmed a particle suspension consistent with an amorphous suspension.Example 11. Preparation of amorphous suspension of anti-PD-1 antibody (pembrolizumab) pH 8.0 I 19.8 % PEG 3350

[0178] Mix 6.66 ml of 50 mM HEPES, pH 8.0 containing 30 % w / v PEG 3350 with 3.33 ml of 50 mg / ml pembrolizumab in 20 mM histidine, pH 5.4 solution at room temperature. The solution was added slowly and mixed after every few drops into the HEPES / PEG solution. The resulting suspension was incubated at room temperature for 2 hours while rotating at 12 rpm on a tube mixer. The amorphous pembrolizumab suspension was concentrated by centrifugation at 3500 RPM in a Beckman Coulter Alegra X-15R centrifuge using a SX-4750 rotor for 1.5 hours (1.5 ml). The resulting pellet was centrifuged in a microfuge at 8000 RPM for 30 minutes (0.70 ml final; 197 mg / ml total). The final buffer composition was 33 mM HEPES, pH 8.0, 19% PEG 3350. The pellet was back loaded into a 2.25 ml Schott glass thin walled staked 27-gauge syringe. The photomicrograph of the anti-PD-1 (197 mg / ml), 33 mMHepes, pH 8.0, 19% PEG 3350 suspension is shown in FIG. 4.Example 12. Preparation of amorphous suspension of aTIGIT (vibostolimab) pH 8.0 / 19.8 % PEG 3350

[0179] Mix 6.66 ml of 50 mM HEPES, pH 8.0 containing 30 % w / v PEG 3350 with 3.33 ml of 70 mg / ml aTIGIT in 20 mM histidine, pH 5.4 solution at room temperature. The solution was added slowly and mixed after every few drops into the HEPES / PEG solution. The resulting suspension was incubated at room temperature for 2 hours while rotating at 12 rpm on a tube mixer. The amorphous aTIGIT suspension was concentrated by centrifugation at 3500 RPM in a Beckman Coulter Alegra X-15R centrifuge using a SX-4750 rotor for 1.5 hours (1.5 ml). The resulting pellet was centrifuged in a microfuge at 8000 RPM for 30 minutes (1.08 ml final; 180 mg / ml total). The final buffer composition was 33 mM HEPES, pH 8.0, 19% PEG 3350. The26059pellet was back loaded into a 2.25 ml Schott glass thin walled staked 27-gauge syringe. The photomicrograph of this example is shown in FIG. 5.Example 13. Preparation of amorphous suspension of aTIGIT (vibostolimab) pH 8.0 / 19.8 % PEG 335019.8 % / NaCl I L-arginine

[0180] Mix 6.66 ml of 50 mM HEPES, pH 8.0 containing 30 % w / v PEG 3350, 50 mM NaCl, 50 mM L-arginine with 3.33 ml of 70 mg / ml aTIGIT (vibostolimab) in 20 mM histidine, pH 5.4 solution at room temperature. The solution was added slowly and mixed after every' few drops into the HEPES / PEG solution. The resulting suspension was incubated at room temperature for 2 hours while rotating at 12 rpm on a tube mixer. The amorphous aTIGIT suspension was concentrated by centrifugation at 3500 RPM in a Beckman Coulter Alegra X-15R centrifuge using a SX-4750 rotor for 1.5 hour (1.3 ml). The resulting pellet was centrifuged in a microfuge at 3000 RPM for 15 minutes. (1.0 ml final; 194 mg / ml total). The final buffer composition was 33 mM HEPES, pH 8.0, 19% PEG 3350, 33mM NaCl, 33 mM arginine. The pellet was back loaded into a 2.25 ml Schott glass thin walled staked 27-gauge syringe. The photomicrograph of this example is shown in FIG. 6.Example 14. Preparation of amorphous suspension of aPD-1 +aTIGIT (vibostolimab) combination formulation pH 8.0 / 19.8 % Z33 mM NaCl Z33 mM L-arginine

[0181] Mix 6.66 ml of 50 mM HEPES, pH 8.0 containing 30 % w / v PEG 3350, 50 mM NaCl, 50 mM arginine with 2.32 ml of 50 mg / ml pembrolizumab in 20 mM histidine, pH 5.4 solution and 1.64 ml of 70 mg / ml aTIGIT (vibostolimab) in 20 mM histidine, pH 5.4 solution at room temperature. The solution was added slowly and mixed after every few drops into the HEPES / PEG solution. The resulting suspension was incubated at room temperature for 2 hours while rotating at 12 rpm on a tube mixer. The amorphous aPD-1 -aTIGIT suspension was centrifuged at 3500 RPM in a Beckman Coulter Alegra X-15R centrifuge using a SX-4750 rotor for 1.5 hour (1.3 ml). The resulting pellet was centrifuged in a microfuge at 3000 RPM for 15 minutes. (1.0 ml final; 192 mg / ml total). The final buffer composition was 33 mM HEPES, pH 8.0, 19% PEG 3350, 33 mM NaCl, 33 mM L-arginine. The pellet was back loaded into a 2.25 ml Schott glass thin walled staked 27-gauge syringe. The photomicrograph of this example is shown in FIG. 7.Injectability method:26059

[0182] A customized experimental framework was developed to gain fundamental insight into the physics of suspension clogging and identify design pathways to prevent clogging risk. The injection setup accommodates vary ing a defined range of parameters across syringeability7tests and was used to measure break-loose and extrusion force. The custom setup provides a range of advantages that includes: (i) driving injection with a controlled rate of plunger motion, (ii) monitoring plunger injection force as a function of time, and (iii) visualizing particle flow throughout the injection time. There were several key features built into the system required to extract useful insight about injection response and potential clogging performance. A standard syringe pump (PHD ULTRATM, Harvard apparatus, Holliston. MA), in the vertical orientation, enabled vertical injections to account for the effect of gravity for consistency with the use-case scenario. 3D-printed fixtures coupled a force sensor (Loadstar Sensors, part# MFD- 100-050-S*C01) to monitor plunger force throughout injection as a first measure of clogging behavior. Additionally, the ability to record fluorescent videos can add a visual inspection mode to bring insight into changes in local particle concentration inside the transparent syringe.

[0183] Vertical injection experiments into air were conducted while monitoring the plunger force using a force sensor. Before each injection experiment, prefilled syringes loaded with the formulation were stoppered leaving an air gap of approximately 2 mm. The plunger rod was then screwed into the prefilled syringe stopper and the needle shield was removed. The prefilled syringe with plunger rod was secured into the injected system and the test was run. Expelled inj ectate was collected in a glass container.Injectability results:Table 6. Measured break-loose force and average extrusion force of various formulations injected using 2.25 mL glass syringe with 27G thin-walled staked-in needle.26059Example 15: Preparation of amorphous suspension of aTIGIT (vibostolimab) pH 8.0119.8 % / 33 mM NaCl I 33 mM L-arginine C13 SSNMR example

[0184] Mix 6.66 ml of 50 mM HEPES, pH 8.0 containing 30 % w / v PEG 335050 mM NaCl, 50 mM L-arginine with 3.33 ml of 70 mg / ml aTIGIT (vibostolimab) in 20 mM histidine, pH 5.4 solution at room temperature. The solution was added slowly and mixed after every few drops into the Hepes / PEG solution. The resulting suspension was incubated at room temperature for 2 hours while rotating at 12 rpm on a tube mixer. The amorphous aTIGIT suspension was centrifuged at 3500 RPM for 1.5 hour (1.3 ml). The resulting pellet was centrifuged in a microfuge at 3000 RPM for 1 minutes. (1.0 ml final; 230 mg / ml total). The final buffer composition was 33 mM Hepes, pH 8.0, 19% PEG 3350, 33 mM NaCl, 33 mM arginine. A sample was shipped for C 13 SSNMR experiments.Solid-state NMR spectroscopy analyses the amorphous suspension of aTIGIT (vibostolimab) pH 8.0119.8 % Z33 mM NaCl I 33 mM L-arginine

[0185] Solid-state NMR spectra are acquired on a Bruker Avance III 400 MHz spectrometer equipped with a 4.0 mm H / F / X magic angle spinning (MAS) probe. The probe was tuned to double resonance C / H for 13C (carbon-13) experiments. The MAS frequency for all experiments is 12 kHz. The sample temperature was controlled at 10°C. 13C cross polarization (CP) magic angle spinning (MAS) spectra were collected under 71.4 kHz 1H dipolar decoupling during acquisition, with a CP contact time of 1 millisecond and a recycle delay of 2 seconds. 13C chemical shifts were referenced to the 13C signal of the carbonyl carbon of glycine at 176.45 ppm.

[0186] Using the solid state 13C 400MHz NMR equipment and procedures described above, the13C (carbon- 13) CP MAS NMR spectra of aTIGIT (vibostolimab) suspension sample have been acquired. The full and enlarged spectral regions exhibiting the resolution 13C peaks are respectively shown in Figure 8A and 8B. This aTIGIT amorphous suspension exhibits generally broad peaks, representing the nature of amorphous phase. However, the spectrum also shows many relatively narrow- peaks, a similar observation as the amorphous suspension of pembrolizumab, suggesting some extent of local order of molecular packing.26059Example 16. Attempted preparation of amorphous suspension of anti-TIGIT (vibostolimab) antibody

[0187] 666 pl of 20 mM histidine, pH 5.4 containing 25 % w / v PEG was mixed with 333 pl of 50 mg / ml aTIGIT (vibostolimab) in 20 mM histidine, pH 5.4 solution at room temperature. Solutions were prepared with distilled water and were filtered with a 0.22-micron filter. The precipitation mixture was kept at room temperature for 4 hours while rotating at 12 rpm on a tube mixer.Example 17. Preparation of amorphous suspension of anti-PD-1 antibody (pembrolizumab)

[0188] 666 pl of 50 mM HEPES, pH 7.0 containing 25 % w / v PEG was mixed with 333 pl of 40 mg / ml pembrolizumab in 20 mM histidine, pH 5.4 solution at RT. Precipitation was carried out in batch mode at total volume of 1 rnL, with each batch yielding -13 mg of the antibody. Solutions were prepared with distilled water and were filtered with a 0.22-micron filter. The precipitation mixture was kept at room temperature for 2 hours while rotating at 12 rpm on a tube mixer. An amorphous pembrolizumab suspension was recovered by centrifugation at 1000 RCF for 5 minutes. A280 UV measurements were made of the resulting pellet suspension (72 mg / ml) and supernatant (0.1 mg / ml) after a 1: 10 dilution in 20 mM Hepes, pH 7.0. SONICC inspection by Visible. UV+ and SHG- imaging confirmed a particle suspension consistent with an amorphous suspension.Example 18. Preparation of amorphous solid forms of anti-RSV antibody (clesrovimab)

[0189] 666 pl of 50 mM HEPES, pH 7.0 containing 25 % w / v PEG was mixed with 333 pl of 40 mg / ml of anti-RSV antibody (clesrovimab) in 20 mM histidine, pH 5.4 solution at room temperature. Precipitation was carried out in batch mode at total volume of 1 mL, yielding -13 mg of the antibody. Solutions were prepared with distilled water and were filtered with a 0.22-micron filter. The precipitation mixture was kept at room temperature for 4 hours while rotating at 12 rpm on a tube mixer. An amorphous anti-RSV antibody (clesrovimab) suspension was recovered by centrifugation at 1000 RCF for 5 minutes. A280 UV measurements were made of the resulting pellet suspension (157 mg / ml) and supernatant (0.1 mg / ml) after a 1:10 dilution in 20 mM Hepes, pH 7.0. SONICC inspection by Visible, UV+ and SHG- imaging confirmed a particle suspension consistent with an amorphous suspension.Example 19. Preparation of amorphous solid forms of anti-RSV antibody (clesrovimab)26059

[0190] 666 (il of 50 mM HEPES, pH 7.0 containing 25 % w / v PEG was mixed with 333 pl of 40 mg / ml of anti-RSV antibody (clesrovimab) in 20 mM histidine, pH 5.4 solution at room temperature. Precipitation was carried out in batch mode at total volume of 1 mL, yielding -13 mg of the antibody. Solutions were prepared with distilled water and were filtered with a 0.22-micron filter. The precipitation mixture was kept at room temperature for 4 hours while rotating at 12 rpm on a tube mixer. An amorphous anti-RSV antibody (clesrovimab) suspension was recovered by centrifugation at 1000 RCF for 5 minutes. A280 UV measurements were made of the resulting pellet suspension (180 mg / ml) and supernatant (0.1 mg / ml) after 1:10 dilution in 20 m Hepes, pH 7.0. SONICC inspection by Visible. UV+ and SHG- imaging confirmed a particle suspension consistent with an amorphous suspension.Example 20. Preparation of co-formulation amorphous suspension of anti-PD-1 antibody (pembrolizumab) and aTIGIT (vibostolimab) in a single suspension

[0191] 666 pl of 50 mM HEPES, pH 8.0 containing 25 % w / v PEG was mixed with 166 pl of 40 mg / ml pembrolizumab in 20 mM histidine, pH 5.4 solution and 166 pl of 70 mg / ml of anti-TIGIT antibody (vibostolimab) in 20 mM histidine, pH 5.4 at room temperature in a single formulation. Precipitation of the formulation comprising both antibodies was carried out in batch mode at total volume of 1 mL. with each batch yielding -13 mg of the antibody (combined). The precipitation mixture was kept at room temperature for 4 hours while rotating at 12 rpm on a tube mixer. The amorphous coformulation suspension was recovered by centrifugation at 1000 RCF for 5 minutes. A280 UV measurements were made of the resulting pellet suspension (57 mg / ml) and supernatant (0.1 mg / ml). SONICC inspection by Visible, UV+ and SHG- imaging confirmed a particle suspension consistent with an amorphous suspension.Example 21. Preparation of co-formulation amorphous suspension of anti-RSV antibody (clesrovimab) and aHMPV antibody in a single suspension

[0192] 666 pl of 50 mM HEPES, pH 7.0 containing 25 % w / v PEG was mixed with 166 pl of 40 mg / ml of anti-RSV antibody (clesrovimab) in 20 mM histidine, pH 5.4 and 166 pl of 70 mg / ml aHMPV in 20 mM sodium glutamate, pH 5.4 solution at room temperature in a single formulation. Precipitation was carried out in batch mode at total volume of 1 mL, with each batch yielding -13 mg of the antibody (combined). The precipitation mixture was kept at room temperature for 4 hours while rotating at 12 rpm on a tube mixer. The resulting amorphous suspension was recovered by centrifugation at 1000 RCF for 5 minutes. A280 UV measurements w ere made of the resulting pellet suspension (83 mg / ml) and supernatant (0.1 mg / ml). SONICC26059inspection by Visible, UV+ and SHG- imaging confirmed a particle suspension consistent with an amorphous suspension.Example 22. Preparation of amorphous suspension of anti-PD-1 antibody (pembrolizumab) micro-ED experiment

[0193] 666 pl of 50 mM HEPES, pH 7.0 containing 25 % w / v PEG was mixed with 333 pl of 40 mg / ml pembrolizumab in 20 mM histidine, pH 5.4 solution at room temperature. Precipitation was carried out in batch mode at total volume of 1 mL, with each batch yielding ~13 mg of the antibody. Solutions were prepared with distilled water and were filtered with a 0.22-micron filter. The precipitation mixture was kept at room temperature for 2 hours while rotating at 12 rpm on a tube mixer. An amorphous pembrolizumab suspension was recovered by centrifugation at 1000 RCF for 5 minutes. A280 UV measurements were made of the resulting pellet suspension (68 mg / ml) and supernatant (0.1 mg / ml) after 1:10 dilution in PBS. SONICC inspection by Visible, UV+ and SHG- imaging confirmed a particle suspension consistent with an amorphous suspension.

[0194] The pellet was resuspended in 50 mM Hepes, 8% PEG 3350. Crystalline aPD-1 was prepared as previously disclosed and amorphous suspensions were diluted 1:10 in 10 mM HEPES (pH 7.0) and 8% PEG 3350 immediate before 3pL was applied to a glow discharged Quantifoil R2 / 2 grid. Grids were plunge frozen using a Leica GP2 and stored in liquid nitrogen until they were loaded into a cryo focused ion beam scanning electron microscope (cryoFIB; Aquilos; Thermo Fisher Scientific). Lamellae were milled in a stepwise fashion to a final thickness of -200 nm. Grids were stored in liquid nitrogen before being imaged using a 200 kV transmission electron microscopy (Talos Arctica; Thermo Fisher). Diffraction and real space images were collected from lamellae containing either the amorphous solution or the crystal form. The cry stal form did not diffract well, nevertheless, a crystalline lattice pattern can be seen in the diffraction images from the crystalline form but not the amorphous form. Further supporting this observation, real space images of the crystal form showed clear unit cell ordering, while the amorphous form did not. Taken together, this suggests that the amorphous form is truly amorphous in nature. See FIG. 9.Example 23: Preparation of amorphous suspension of anti-PD-1 antibody (pembrolizumab) for SSNMR experiment

[0195] 0.5 ml of 100 mM HEPES, pH 8.2 containing 30 % w / v PEG was mixed with 0.5 ml of 225 mg / ml pembrolizumab in 20 mM histidine, pH 5.4 solution at room temperature.26059Precipitation was carried out in batch mode at total volume of 1 mL, with each batch yielding -13 mg of the antibody. The precipitation mixture was kept at room temperature for 4 hours while rotating at 12 rpm on a tube mixer. An amorphous pembrolizumab suspension was recovered by centrifugation at 3000 RCF for 15 minutes. The supernatant was removed. The resulting suspension was sent for SSNMR analyses. SONICC imaging results shown in Figure 10 by Visible, UV+ and SHG- imaging confirmed a particle suspension consistent with an amorphous suspension.Example 24. Solid-state NMR spectroscopy analyses crystalline, amorphous, liquid and lyophilized aPD-1 (pembrolizumab)

[0196] Solid-state NMR spectra are acquired on a Bruker Avance III 500 MHz spectrometer equipped with a 4.0 mm H / X / Y magic angle spinning (MAS) probe. The probe was tuned to double resonance C / H for13C (carbon- 13) experiments. The MAS frequency for all experiments is 12 kHz. The sample temperature was controlled at 10 °C.13C cross polarization (CP) MAS spectra are collected under 71.4 kHz 'H dipolar decoupling during acquisition, with a CP contact time of 1 millisecond and a recycle delay of 2 seconds.13C chemical shifts are referenced to the13C signal of the carbonyl carbon of glycine (a-form) at 176.45 ppm.

[0197] Using the solid state13C 500MHz NMR equipment and procedures described above, four pembrolizumab samples have been measured including pembrolizumab crystalline suspension, amorphous suspension, liquid high concentration pembrolizumab formulation, and lyophilized pembrolizumab formulation. Thel3C (carbon-13) CP MAS NMR spectra of the four samples were obtained. The full and enlarged spectral regions exhibiting the resolution13C peaks are respectively shown in Figure 11A and B. The lyophilized pembrolizumab formulation exhibits broad peaks, representing the conformational inhomogeneity7. The liquid high concentration pembrolizumab formulation demonstrates a broad baseline, resulting from slow- molecular tumbling in the highly viscous solution. The amorphous suspension exhibited generally broader peaks than the pembrolizumab caffeine crystalline suspension, representing the amorphous nature. However, there were many relatively narrow peaks in the spectrum of the amorphous suspension, suggesting some extent of local order of molecular packing.Example 25: Preparation of amorphous suspension of anti-PD-1 antibody (pembrolizumab)

[0198] 12 ml of 35 mM HEPES, pH 7.0 containing 25 % w / v PEG, 28 mM arginine was mixed with 6 ml of 40 mg / ml pembrolizumab in 20 mM histidine, pH 5.4 solution at room temperature. Precipitation w as carried out in batch mode at a total volume of 18 mL. The precipitation mixture26059was kept at room temperature for 4 hours. The vial was mixed on a MediMixlOO® automated mixer (Health mark, CA) set at the lowest speed at room temperature. The suspension was observed to change to a white paste slowly within 2 hours at room temperature. The amorphous pembrolizumab suspension was recovered by centrifugation in a Beckman Coulter centrifuge Allegra X-15R at 3500 RPM for 2 hours at 4°C. A280 UV measurements were made of the resulting pellet suspension (195 mg / ml) and supernatant (0.1 mg / ml). SONICC inspection by Visible, UV+ and SHG- imaging confirmed a particle suspension consistent with an amorphous suspension.Example 26. HP-SEC characterization data

[0199] The aggregation propensity of amorphous suspensions of different antibodies was analyzed by Ultra-high-performance size exclusion chromatography (UPSEC) following reconstitution into the histidine buffer centrifugation at 10000 rpm. UPSEC was performed on a Waters Acquity UPLC system using a Waters BEH200 SEC column (4.6 x 300mm, 1.7pm) according to the method described previously (Mou et al., 2014) with modified mobile phase consisting of 50mM phosphate and 450mM Arginine HC1, pH7.0. The sampler temperature was 5°C, the column temperature was 30.0°C. Typically, 30 mg of sample was injected and run for 25 min at a flow rate of 0.2 mL / min. The elution was monitored by UV absorbance at 280 nm.

[0200] UPSEC data clearly showed that no appreciable increase in HMW was observed for different antibodies tested after formation of amorphous suspensions using PEG-3350, and the majority of antibodies remained as monomer. These data suggest that the confirmation of antibodies is not changed to induce the protein-protein interactions, including formation of aggregates.Table 7: USPEC data26059Example 27A: Preparation of amorphous solid forms of ROR1-ADC

[0201] 66 pl of 50 mM HEPES, pH 7.0 containing 25 % w / v PEG was mixed with 33 pl of 50 mg / ml of ROR1-ADC in 20 mM histidine, pH 5.4 solution at room temperature. Precipitation was carried out in batch mode at total volume of 0.1 mL, yielding ~1.7 mg of the ADC. Solutions were prepared with distilled water and were filtered with a 0.22-micron filter. The precipitation mixture was kept at room temperature for 4 hours while rotating at 12 rpm on a tube mixer. An amorphous suspension was recovered by centrifugation at 1000 RCF for 5 minutes. A280 UV measurements were made of the resulting pellet suspension (60 mg / ml) and supernatant (0.1 mg / ml) after 1:10 dilution in 20 mM Hepes. pH 7.0. SONICC inspection by Visible, UV+ and SHG- imaging confirmed a particle suspension consistent with an amorphous suspension. See FIG. 12.Example 27B: Preparation of amorphous solid forms of aTau antibody

[0202] 66 pl of 50 mM HEPES, pH 7.0 containing 30 % w / v PEG was mixed with 33 pl of 150 mg / ml of the aTau antibody in 20 mM histidine, pH 5.4 solution at room temperature.Precipitation was carried out in batch mode at total volume of 0.1 mL, yielding -4 mg of the ADC. Solutions were prepared with distilled water and were filtered with a 0.22-micron filter. The precipitation mixture was kept at room temperature for 4 hours while rotating at 12 rpm on a tube mixer. An amorphous suspension was recovered by centrifugation at 1000 RCF for 5 minutes. A280 UV measurements were made of the resulting pellet suspension (73 mg / ml) and supernatant (0.1 mg / ml) after 1:10 dilution in 20 mM Hepes, pH 7.0. SONICC inspection by Visible, UV+ and SHG- imaging confirmed a particle suspension consistent with an amorphous suspension. See FIG. 13.Example: 28A: Preparation of pembrolizumab suspension samples for rheology and Solid-State NMR experiments

[0203] 3.3 ml of pembrolizumab (50 mg / ml) in 20 mM histidine, pH 6 was added to 6.6 ml of 50 mM Hepes, pH 7.4, 30 % PEG 3350 at RT and allowed to rotate on a rotator for 2 hours. The resulting suspension was centrifuged at 1 OK RPM for 20 minutes. The resulting supernatant was removed from the white solid pellet. A260 / 280 nm UV was 0.014 mg fin. The resulting amorphous suspension was 200 mg / ml (0.75 ml) in 33 mM Hepes. pH 7.4, 20 % PEG 3350. FIGs. 14A and 14B show photo micrographs at 200X magnification of the pre and post rheology and SSNMR images.26059Example 28B: Amorphous pembrolizumab suspensions rheology experiment

[0204] Rheometry data was acquired on a Discovery HR-20 rheometer equipped with a 40 mm sandblasted parallel plate geometry. The temperature was set to 20°C. Samples were pre-sheared at a shear rate of 10 s'1for 30 seconds to erase the sample history prior to measurements. The viscosity (77) was then measured as a function of shear rate (y) ranging from 0.1 s’1to 2000 s’1, wi th steady state defined as the value for which the shear stress measured across three consecutive 10-second intervals were within 5% of each other.

[0205] Using the equipment and procedures described above, an amorphous pembrolizumab suspension sample was characterized. The steady-state viscosity as a function of the shear rate is shown in Figure 15 A. The data collected from the viscosity measurements can also be assessed in terms of the equilibrium total strain, which is the product of the shear rate and the length of time for which the shear rate was applied. The equilibrium total strain plotted as a function of the shear rate is shown in Figure 15B. The viscosity functions were further examined by Sisko viscosity model (Equation 1). To achieve 25 cP, pembrolizumab needs 3330 s’1. Using Figure 15B and Equation 2, the time (t) required to have equilibrium total strain for the target 25 cP can be estimated, which is 18 and 23 minutes, respectively. The model parameters are listed in Table 8. Using injection force model from A. Allmendinger et al. (Allmendinger A, et al. "‘Rheological characterization and injection forces of concentrated protein formulations: An alternative predictive model for non-Newtonian solutions.” European Journal of Pharmaceutics and Biopharmaceutics 2014;87(2):318-328) and shear-thinning viscosity7curve, the measurement suggests a glide force of 16N.Equation 1:77 = 7700 + Aq / ”-1Where 77 is the infinite viscosity7, K and n are model parameters.Equation 2:Where A, B. and c are model parameters.Table 8: Model parameters for the amorphous pembrolizumab suspensions rheology experiment26059Example 28C: Solid-state NMR spectroscopy analyses for the amorphous suspension of pembrolizumab

[0206] Solid-state NMR spectra were acquired on a Bruker NEO 500 MHz spectrometer equipped with a 4.0 mm H / F / X magic angle spinning (MAS) probe. The probe was tuned to double resonance C / H for13C (carbon- 13) experiments. The MAS frequency for all experiments was 12 kHz. The sample temperature was controlled at 5°C.13C cross polarization (CP) MAS spectra were collected under 100kHz 'H dipolar decoupling during acquisition, with a CP contact time of 2 millisecond and a recycle delay of 3 seconds.13C chemical shifts were referenced to the methine carbon of adamantane signal at 38.48 ppm.

[0207] Using the solid state13C 500MHz NMR equipment and procedures described above, the13C (carbon- 13) CP MAS NMR spectra of pembrolizumab (aPDl) suspension sample before and after rheology measurements have been acquired and are shown in FIGs. 16A and 16B.

[0208] Figure 16A is ID13C spectra of the amorphous suspension of pembrolizumab (aPDl) at 200mg / mL, collected before (upper panel) and after (lower panel) rheology. FIG. 16B shows the high degree of similarity between the two samples.

[0209] The disclosed subject matter is not to be limited in scope by the specific embodiments and examples described herein. Indeed, various modifications of the disclosure in addition to those described will become apparent to those skilled in the art from the foregoing description and accompanying figures. Such modifications are intended to fall within the scope of the appended claims.

[0210] All references (e g., publications or patents or patent applications) cited herein are incorporated herein by reference in their entirety and for all purposes to the same extent as if each individual reference (e.g.. publication or patent or patent application) was specifically and individually indicated to be incorporated by reference in its entirety for all purposes. Other embodiments are within the following claims.

Claims

WHAT IS CLAIMED IS:

1. A biologic suspension comprising:a) about 1 mg / ml to about 400 mg / ml of a first biologic active ingredient, wherein the first biologic active ingredient is an anti-PDl antibody;b) about 5 mM to about 100 mM buffer; andc) about 5% to about 30% polyethylene glycol (PEG);wherein the suspension has a pH between 5.0 and 9.0 and is amorphous.

2. The biologic suspension of claim 1, wherein the PEG is PEG 3350.

3. The biologic suspension of claim 2, wherein the concentration of PEG 3350 is 10 mg / ml.

4. The biologic suspension of any preceding claim, wherein the molecular weight of the PEG is from about 2,500 to about 20,000.

5. The biologic suspension of claim 1, wherein the PEG is present in an amount of 15 % weight / volume (w / v).

6. The biologic suspension of claim 1 , wherein the PEG is present in an amount of 25 % weight / volume (w / v).

7. The biologic suspension of any preceding claim, wherein the buffer is HEPES, a Tris buffer, a phosphate buffer, MES, or MOPS.

8. The biologic suspension of any preceding claim, wherein the buffer is HEPES.

9. The biologic suspension of claim 8, wherein the HEPES buffer is present at a concentration of about 5-100 mM.

10. The biologic suspension of claim 9, wherein the HEPES buffer is present at a concentration of about 5-75 mM.

11. The biologic suspension of claim 10, wherein the HEPES buffer is present at a concentration of about 10-50 mM.

12. The biologic suspension of claim 11, wherein the HEPES buffer is present at a concentration of about 10 mM.

13. The biologic suspension of claim 11, wherein the HEPES buffer is present at a concentration of about 50 mM.

14. The biologic suspension of any preceding claim, wherein the suspension further comprises histidine.

15. The biologic suspension of claim 14, wherein the histidine concentration is about 20 mM.

16. The biologic suspension of any preceding claim, wherein the suspension further comprises arginine.

17. The biologic suspension of claim 16, wherein the arginine concentration is about 20-50 mM.

18. The biologic suspension of claim 17, wherein the arginine concentration is about 50 mM.

19. The biologic suspension of any preceding claim, further comprising about 6% to about 8% weight / volume (w / v) sucrose, trehalose or (2- hydroxypropyl)-0-cyclodextrin.

20. The biologic suspension of any preceding claim, wherein the suspension further comprises about 6% to about 8% weight / volume (w / v) sucrose.

21. The biologic suspension of any preceding claim, wherein the suspension further comprises sodium chloride.

22. The biologic suspension of claim 21, wherein the sodium chloride concentration is 150 mM.

23. The biologic suspension of any preceding claim, wherein the suspension has a pH between 6.2 and 8.2.

24. The biologic suspension of any preceding claim, wherein the suspension has a pH of 7.0.

25. The biologic suspension of claim 1, wherein the suspension has a pH between 7.0 and 9.0.

26. The biologic suspension of claim 25, wherein the suspension has a pH between 7.0 and 8.5.

27. The biologic suspension of claim 26, wherein the suspension has a pH of 7.0.

28. The biological suspension of claim 1, wherein the pH is between 6.8 and 9.0.

29. The biologic suspension of any one of claims 1-28, wherein the anti -human PD-1 antibody comprises three light chain CDRs comprising a CDRL1 comprising the amino acid sequence of SEQ ID NO: 1, a CDRL2 comprising the amino acid sequence of SEQ ID NO:2 and a CDRL3 comprising the amino acid sequence of SEQ ID NO:3 and three heavy chain CDRs comprising a CDRH1 comprising the amino acid sequence of SEQ ID NO: 6, a CDRH2 comprising the amino acid sequence of SEQ ID NO: 7 and a CDRH3 comprising the amino acid sequence of SEQ ID NO: 8.

30. The biologic suspension of any one of claims 1-29, wherein the anti-PD-1 antibody is pembrolizumab.

31. The biologic suspension of any one of claims 1-30, wherein the biologic suspension further comprises a second biologic active ingredient.

32. The biologic suspension of claim 31, wherein the second biologic active ingredient comprises an anti -TI GIT antibody.

33. The biologic suspension of claim 31, wherein the second biologic active ingredient comprises vibostolimab.2605934. The biologic suspension of claim 32, wherein the biologic suspension comprises 260 mg of the anti-TIGIT antibody and 260 mg of the pembrolizumab.

35. The biologic suspension of claim 34, wherein the biologic suspension comprises an anti-TIGIT antibody and pembrolizumab in a single injection.

36. The biologic suspension of any one of claims 1-30, wherein the biologic suspension was produced by a method comprising:a) mixingi. an aqueous buffered solution comprising the anti-PDl antibody, and ii. about 5 mM to about 100 mM buffer, wherein the buffer is selected from HEPES, a Tris buffer, a phosphate buffer, MES, and MOPS;to form a mixture; andb) precipitating the mixture with PEG at an incubation temperature of about 2°C to about 40°C to form an amorphous suspension;wherein the biologic suspension has a pH above 6 and is amorphous; and wherein the precipitation is performed by vapor diffusion, dialysis, or batch mode.

37. The biologic suspension of any one of claims 1-36, wherein the biologic suspension is encapsulated into a delivery system.

38. The biologic suspension of any one of claims 1-37, wherein the delivery system is a patch or hydrogel.

39. The biologic suspension of claim 38, wherein the delivery system is a hydrogel.

40. The biologic suspension of claim 38, wherein the delivery system is a patch.

41. A method for producing a biologic amorphous suspension comprising:a) mixingi. an aqueous buffered solution comprising a first biologic active ingredient wherein the first biologic active ingredient is an anti-PDl antibody, and ii. about 5 mM to about 100 mM buffer, wherein the buffer is selected from HEPES, a Tris buffer, a phosphate buffer, MES, and MOPS;to form a mixture; andb) precipitating the mixture with PEG at an incubation temperature of about 2°C to about 40°C to form an amorphous suspension;wherein the suspension has a pH between 5 and 9 and is amorphous; and wherein the precipitation is performed by vapor diffusion, dialysis, or batch mode.

42. The method of claim 41, wherein the first biologic active ingredient is pembrolizumab.

43. The method of claim 41 or claim 42. wherein the buffer is histidine.

44. The method of any one of claims 41-43, wherein the concentration of the anti-PDl antibody is about 1 mg / ml to about 400 mg / ml.

45. The method of any one of claims 41-44, wherein the concentration of the PEG is about 5% to 30%.

46. The method of any one of claims 41-45, wherein the pH of the biologic suspension is between 6 and 9.

47. The method of claim 46, wherein the pH of the biologic suspension is between 6.8 and 9.

48. The method of any one of claims 41-47, wherein the PEG is PEG 3350.

49. The method of claim 48, wherein the concentration of PEG 3350 is 10 mg / ml.

50. The method of any one of claims 41-49, wherein the molecular weight of the PEG is from about 2,500 to about 20,000.

51. The method of any one of claims 41-50, wherein the PEG is present in an amount of 15 % weight / volume (w / v).

52. The method of claim 51, wherein the PEG is present in an amount of 25 % weight / volume (w / v).

53. The method of any one of claims 41-52, wherein the buffer is HEPES.

54. The method of any one of claims 41-53, wherein the buffer is about 5-100 mM HEPES.

55. The method of claim 54, wherein the buffer is about 5-75 mM HEPES.

56. The method of claim 55, wherein the buffer is about 10-50 mM HEPES.

57. The method of claim 56, wherein the buffer is about 10 mM HEPES.

58. The method of claim 57, wherein the buffer is about 50 mM HEPES.

59. The method of any one of claims 41-58, wherein the biologic amorphous suspension further comprises histidine.

60. The method of claim 59, wherein the histidine concentration is about 20 mM.

61. The method of any one of claims 41-60. wherein the biologic amorphous suspension further comprises arginine.

62. The method of claim 61, wherein the biologic amorphous suspension comprises about 20-50 mM arginine.

63. The method of claim 62, wherein the arginine concentration is about 50 mM.

64. The method of any one of claims 41-63, wherein the suspension further comprises about 6% to about 8% weight / volume (w / v) sucrose, trehalose or (2- hydroxypropyl)-P-cyclodextrin.

65. The method of claim 64, wherein the suspension comprises about 6% to about 8% weight / volume (w / v) sucrose.

66. The method of any one of claims 41-65, w herein the suspension further comprises sodium chloride.

67. The method of claim 66, wherein the suspension comprises about 150 mM sodium chloride.

68. The method of any one of claims 41-67, wherein the suspension has a pH between 6.2 and 8.2.

69. The method of any one of claims 41-67, wherein the suspension has a pH between 7.0 and 9.0.

70. The method of claim 69, wherein the suspension has a pH between 7.0 and 8.5.

71. The method of claim 70, wherein the suspension has a pH of 7.0.

72. The method of any one of claims 41-67, wherein the biologic amorphous suspension has a pH between 6.8 and 9.0.

73. The method of any one of claims 41-72, wherein the anti-human PD-1 antibody comprises three light chain CDRs comprising a CDRL1 comprising the amino acid sequence of SEQ ID NO: 1 , a CDRL2 comprising the amino acid sequence of SEQ ID NO:2 and a CDRL3 comprising the amino acid sequence of SEQ ID NO: 3 and three heavy chain CDRs comprising a CDRH1 comprising the amino acid sequence of SEQ ID NO:6, a CDRH2 comprising the amino acid sequence of SEQ ID NO:7 and a CDRH3 comprising the amino acid sequence of SEQ ID NO:8.

74. The method of claim 73, wherein the anti-PD-1 antibody is pembrolizumab.

75. The method of any one of claims 41-74, wherein the biologic suspension further comprises a second biologic active ingredient.

76. The method of claim 75, wherein the second biologic active ingredient comprises an anti-TIGIT antibody.

77. The method of claim 75, wherein the second biologic active ingredient biologic comprises vibostolimab.

78. The method of claim 76, wherein biologic amorphous suspension comprises 260 mg of an anti-TIGIT antibody and 260 mg of pembrolizumab.

79. The method of claim 78, wherein biologic suspension comprises an anti-TIGIT antibody and pembrolizumab in a single injection device.

80. The method of any one of claims 41-79, wherein the biologic suspension is encapsulated into a delivery system.

81. The method of claim 80, wherein the delivery system is a patch or hydrogel.

82. The method of claim 81, wherein the delivery system is a hydrogel.

83. The method of claim 81, w herein the delivery system is a patch.

84. A biologic suspension comprising:a) about 5 mg / ml to about 400 mg / ml of pembrolizumab;b) about 5 mM to about 100 mM buffer;c) about 5% to about 30% polyethylene glycol (PEG);d) sodium chloride;e) sucrose;wherein the suspension is amorphous; and wherein the suspension is made by a method comprising:i) mixingi. an aqueous buffered solution comprising about 200 mg of pembrolizumab in histidine; andii. about 5 mM to about 100 mM buffer, wherein the buffer is selected from HEPES, a Tris buffer, a phosphate buffer, MES, and MOPS;to form a mixture; andii) precipitating the mixture with PEG at an incubation temperature of about 2°C to about 40°C to form an amorphous suspension; wherein precipitation is by vapor diffusion, dialysis, or batch mode; and wherein the suspension has a pH between 5.0 and 9.0.

85. A biologic suspension comprising:a) about 5 mg / ml to about 400 mg / ml of pembrolizumab;b) about 5 mM to about 100 mM buffer;c) about 5% to about 30% polyethylene glycol (PEG);d) sodium chloride;e) sucrose;wherein the suspension is amorphous; and wherein the suspension is made by a method comprising:i) mixingi. an aqueous buffered solution comprising about 200 mg of pembrolizumab in histidine; andii. about 5 mM to about 100 mM buffer, wherein the buffer is selected from HEPES, a Tris buffer, a phosphate buffer, MES. and MOPS;to form a mixture; andii) precipitating the mixture with PEG at an incubation temperature of about 2°C to about 40°C to form an amorphous suspension; wherein precipitation is by vapor diffusion, dialysis, or batch mode; and wherein the suspension has a pH between 5.0 and 9.0; andwherein the biologic suspension additionally comprises about 260 mg of vibostolimab.

86. A method of generating a biologic amorphous suspension, the method comprising:a) mixing together a first solution with a second solution to create a third solution, whereini. the first solution comprises an anti-PDl antibody and histidine, wherein the concentration of the anti-PDl antibody in the first solution is between 45 mg / ml and 55 mg / ml, andwherein the concentration of histidine in the first solution is between 18 mM and 22 mM. andwherein the first solution has a pH between 5.5 and 6.5;andii. the second solution comprises a buffer and PEG.wherein the concentration of the buffer in the second solution is between 45 mM and 55 mM, andwherein the concentration of PEG in the second solution is between 25% and 35%;b) centrifuging the third solution to create a supernatant and a pellet,wherein the supernatant is the biologic amorphous suspension; andc) separating the biologic amorphous suspension from the pellet.

87. The method of claim 86, wherein the concentration of the anti-PDl antibody in the first solution is about 50 mg / ml.

88. The method of claim 86 or 87, wherein the anti-human PD-1 antibody comprises three light chain CDRs comprising a CDRL1 comprising the amino acid sequence of SEQ ID NO:1, a CDRL2 comprising the amino acid sequence of SEQ ID NO:2 and a CDRL3 comprising the amino acid sequence of SEQ ID NO:3 and three heavy chain CDRs comprising a CDRH1 comprising the amino acid sequence of SEQ ID NO: 6, a CDRH2 comprising the amino acid sequence of SEQ ID NO:7 and a CDRH3 comprising the amino acid sequence of SEQ ID NO:8.

89. The method of claim 88, wherein the anti-PDl antibody comprises pembrolizumab.

90. The method of claim 86, wherein the concentration of histidine in the first solution is about 20 mM.

91. The method of claim 86, wherein the concentration of the buffer in the second solution is about 50 mM.

92. The method of claim 86 or claim 91, wherein the buffer in the second solution is Hepes buffer.

93. The method of claim 86, wherein the concentration of PEG in the second solution is about 30%.

94. The method of claim 86 or claim 93, wherein the PEG is PEG 3350.

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