Measuring Aggregates, Self-association, and Weak Interactions in Concentrated Therapeutic Antibody Solutions

Overview

Journal MAbs

Specialty Allergy & Immunology

Date 2020 Sep 5

PMID 32887536

Citations 11

Authors

Sumit K Chaturvedi

Arun Parupudi

Kristian Juul-Madsen

Ai Nguyen

Thomas Vorup-Jensen

Sonia Dragulin-Otto

Huaying Zhao

Reza Esfandiary

Peter Schuck

Affiliations

Soon will be listed here.

Abstract

Monoclonal antibodies are a class of biotherapeutics used for an increasing variety of disorders, including cancer, autoimmune, neurodegenerative, and viral diseases. Besides their antigen specificity, therapeutic use also mandates control of their solution interactions and colloidal properties in order to achieve a stable, efficacious, non-immunogenic, and low viscosity antibody solution at concentrations in the range of 50-150 mg/mL. This requires characterization of their reversible self-association, aggregation, and weak attractive and repulsive interactions governing macromolecular distance distributions in solution. Simultaneous measurement of these properties, however, has been hampered by solution nonideality. Based on a recently introduced sedimentation velocity method for measuring macromolecular size distributions in a mean-field approximation for hydrodynamic interactions, we demonstrate simultaneous measurement of polydispersity and weak and strong solution interactions in a panel of antibodies with concentrations up to 45 mg/mL. By allowing approximately an order of magnitude higher concentrations than previously possible in sedimentation velocity size distribution analysis, this approach can substantially improve efficiency and sensitivity for characterizing polydispersity and interactions of therapeutic antibodies at or close to formulation conditions.

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References

Gabrielson J, Randolph T, Kendrick B, Stoner M . Sedimentation velocity analytical ultracentrifugation and SEDFIT/c(s): limits of quantitation for a monoclonal antibody system. Anal Biochem. 2006; 361(1):24-30. DOI: 10.1016/j.ab.2006.11.012. View

Ghirlando R, Balbo A, Piszczek G, Brown P, Lewis M, Brautigam C . Improving the thermal, radial, and temporal accuracy of the analytical ultracentrifuge through external references. Anal Biochem. 2013; 440(1):81-95. PMC: 3826449. DOI: 10.1016/j.ab.2013.05.011. View

Chaturvedi S, Sagar V, Zhao H, Wistow G, Schuck P . Measuring Ultra-Weak Protein Self-Association by Non-ideal Sedimentation Velocity. J Am Chem Soc. 2019; 141(7):2990-2996. PMC: 6385077. DOI: 10.1021/jacs.8b11371. View

Blanco M, Hatch H, Curtis J, Shen V . Evaluating the Effects of Hinge Flexibility on the Solution Structure of Antibodies at Concentrated Conditions. J Pharm Sci. 2018; 108(5):1663-1674. PMC: 7288351. DOI: 10.1016/j.xphs.2018.12.013. View

Blanco M, Sahin E, Li Y, Roberts C . Reexamining protein-protein and protein-solvent interactions from Kirkwood-Buff analysis of light scattering in multi-component solutions. J Chem Phys. 2011; 134(22):225103. PMC: 3133569. DOI: 10.1063/1.3596726. View

Geng S, Cheung J, Narasimhan C, Shameem M, Tessier P . Improving monoclonal antibody selection and engineering using measurements of colloidal protein interactions. J Pharm Sci. 2014; 103(11):3356-3363. PMC: 4206634. DOI: 10.1002/jps.24130. View

Zhao H, Ghirlando R, Piszczek G, Curth U, Brautigam C, Schuck P . Recorded scan times can limit the accuracy of sedimentation coefficients in analytical ultracentrifugation. Anal Biochem. 2013; 437(1):104-8. PMC: 3676908. DOI: 10.1016/j.ab.2013.02.011. View

Moussa E, Panchal J, Moorthy B, Blum J, Joubert M, Narhi L . Immunogenicity of Therapeutic Protein Aggregates. J Pharm Sci. 2016; 105(2):417-430. DOI: 10.1016/j.xphs.2015.11.002. View

Saluja A, Fesinmeyer R, Hogan S, Brems D, Gokarn Y . Diffusion and sedimentation interaction parameters for measuring the second virial coefficient and their utility as predictors of protein aggregation. Biophys J. 2010; 99(8):2657-65. PMC: 2955502. DOI: 10.1016/j.bpj.2010.08.020. View

10.

Some D . Light-scattering-based analysis of biomolecular interactions. Biophys Rev. 2013; 5(2):147-158. PMC: 3641300. DOI: 10.1007/s12551-013-0107-1. View

11.

Calero-Rubio C, Saluja A, Roberts C . Coarse-Grained Antibody Models for "Weak" Protein-Protein Interactions from Low to High Concentrations. J Phys Chem B. 2016; 120(27):6592-605. DOI: 10.1021/acs.jpcb.6b04907. View

12.

Esfandiary R, Parupudi A, Casas-Finet J, Gadre D, Sathish H . Mechanism of reversible self-association of a monoclonal antibody: role of electrostatic and hydrophobic interactions. J Pharm Sci. 2014; 104(2):577-86. DOI: 10.1002/jps.24237. View

13.

Roberts C . Protein aggregation and its impact on product quality. Curr Opin Biotechnol. 2014; 30:211-7. PMC: 4266928. DOI: 10.1016/j.copbio.2014.08.001. View

14.

Jimenez M, Rivas G, Minton A . Quantitative characterization of weak self-association in concentrated solutions of immunoglobulin G via the measurement of sedimentation equilibrium and osmotic pressure. Biochemistry. 2007; 46(28):8373-8. DOI: 10.1021/bi7005515. View

15.

Uchiyama S, Noda M, Krayukhina E . Sedimentation velocity analytical ultracentrifugation for characterization of therapeutic antibodies. Biophys Rev. 2017; 10(2):259-269. PMC: 5899733. DOI: 10.1007/s12551-017-0374-3. View

16.

Luo H, Macapagal N, Newell K, Man A, Parupudi A, Li Y . Effects of salt-induced reversible self-association on the elution behavior of a monoclonal antibody in cation exchange chromatography. J Chromatogr A. 2014; 1362:186-93. DOI: 10.1016/j.chroma.2014.08.048. View

17.

Schuck P, Rossmanith P . Determination of the sedimentation coefficient distribution by least-squares boundary modeling. Biopolymers. 2000; 54(5):328-41. DOI: 10.1002/1097-0282(20001015)54:5<328::AID-BIP40>3.0.CO;2-P. View

18.

Hopkins M, Lambert C, Bee J, Parupudi A, Bain D . Determination of Interaction Parameters for Reversibly Self-Associating Antibodies: A Comparative Analysis. J Pharm Sci. 2018; 107(7):1820-1830. DOI: 10.1016/j.xphs.2018.03.011. View

19.

Skar-Gislinge N, Ronti M, Garting T, Rischel C, Schurtenberger P, Zaccarelli E . A Colloid Approach to Self-Assembling Antibodies. Mol Pharm. 2019; 16(6):2394-2404. DOI: 10.1021/acs.molpharmaceut.9b00019. View

20.

S Sarangapani P, Weaver J, Parupudi A, Besong T, Adams G, Harding S . Both Reversible Self-Association and Structural Changes Underpin Molecular Viscoelasticity of mAb Solutions. J Pharm Sci. 2016; 105(12):3496-3506. DOI: 10.1016/j.xphs.2016.08.020. View