Early determination of potential critical quality attributes of therapeutic antibodies in developability studies through surface plasmon resonance-based relative binding activity assessment

ABSTRACT

Precise measurement of the binding activity changes of therapeutic antibodies is important to determine the potential critical quality attributes (CQAs) in developability assessment at the early stage of antibody development. Here, we report a surface plasmon resonance (SPR)-based relative binding activity method, which incorporates both binding affinity and binding response and allows us to determine relative binding activity of antibodies with high accuracy and precision. We applied the SPR-based relative binding activity method in multiple forced degradation studies of antibody developability assessment. The current developability assessment strategy provided comprehensive, precise characterization of antibody binding activity in the stability studies, enabling us to perform correlation analysis and establish the structure–function relationship between relative binding activity and quality attributes. The impact of a given quality attribute on binding activity could be confidently determined without isolating antibody variants. We identified several potential CQAs, including Asp isomerization, Asn deamidation, and fragmentation. Some potential CQAs affected binding affinity of antibody and resulted in a reduction of binding activity. Certain potential CQAs impaired antibody binding to antigen and led to a loss of binding activity. A few potential CQAs could influence both binding affinity and binding response and cause a substantial decrease in antibody binding activity. Specifically, we identified low abundance Asn33 deamidation in the light chain complementarity-determining region as a potential CQA, in which all the stressed antibody samples showed Asn33 deamidation abundances ranging from 4.2% to 27.5% and a mild binding affinity change from 1.76 nM to 2.16 nM.

KEYWORDS:

• Antibody development
• critical quality attribute
• developability assessment
• forced degradation
• relative binding activity
• surface plasmon resonance
• therapeutic antibody

Acknowledgments

We thank all the members from the biopharmaceutical discovery who helped us by providing the antibodies and antigens for method development. We thank Dr. Zhuoxiao Cao for her support and helpful discussion. We thank the anonymous reviewers for their insightful comments and suggestions.

Disclosure statement

No potential conflict of interest was reported by the author(s).

Author contributions

S.W. designed the study and analyzed the data. Y.W. and Z.L. performed most experiments with help from Y.H., Z.W., and J.F. Q.W., Y.G., X.Z., G.C., C.C., and Y.W. provided antibodies. S.W. and Y.F. wrote the paper.

Data availability statement

All the experimental data that support the findings of this study are presented in the main text and the supplemental material. For more information please contact to the corresponding author Shuai Wang.

Supplementary material

Supplemental data for this article can be accessed online at https://doi.org/10.1080/19420862.2024.2374607

Correction Statement

This article has been corrected with minor changes. These changes do not impact the academic content of the article.

Additional information

Funding

This work was funded by Simcere Zaiming Pharmaceutical.