Monoclonal Antibody Aggregation Intermediates Visualized by Atomic Force Microscopy

• Published in: Journal of pharmaceutical sciences Vol. 100; no. 2; pp. 416 - 423
• Main Authors: Lee, Hanjoo, Kirchmeier, Marc, Mach, Henryk
• Format: Journal Article
• Language: English
• Published: Hoboken Elsevier Inc 01.02.2011; Wiley Subscription Services, Inc., A Wiley Company; Wiley; American Pharmaceutical Association; Elsevier Limited
• Subjects: analytical biochemistry; bioanalysis; Biological and medical sciences; diffusion; General pharmacology; Humans; Immunoglobulin G - chemistry; immunology; injectables; Medical sciences; Microscopy, Atomic Force; Pharmaceutical technology. Pharmaceutical industry; Pharmacology. Drug treatments; protein binding; Protein Conformation; protein delivery; Protein Folding; protein folding/refolding; protein formulation; protein structure; analytical biochemistry; protein structure; protein formulation; protein folding/refolding; injectables; diffusion; protein delivery; bioanalysis; protein binding; immunology; Atomic force microscopy; Pharmaceutical technology; Monoclonal antibody; Aggregation; Binding protein; Formulation; Diffusion
• ISSN: 0022-3549; 1520-6017
• DOI: 10.1002/jps.22279

Ubiquitous but highly variable processes of therapeutic protein aggregation remain poorly characterized, especially in the context of common infusion reactions and clinical immunogenicity. Among the numerous challenges is the characterization of intermediate steps that lead to the appearance of precipitates. Although the biophysical methods for elucidation of secondary and tertiary structures as well as overall size distribution are typically well established in the development laboratories, the use of molecular scale imaging techniques is still relatively rare due to low throughput and technical complexity. In this work, we present the use of atomic force microscopy to examine morphology of monoclonal antibody aggregates. Despite varying in primary structure as a result of different complementarity defining regions, most antibodies studied exhibited a similar aggregation intermediate consisting of several monomers. However, the manner of subsequent condensation of these oligomers appeared to differ between the antibodies, suggesting stability-dependent mechanisms.