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Updated: Jun 19, 2026

Determination of High-affinity Antibody-antigen Binding Kinetics Using Four Biosensor Platforms
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Published on: April 17, 2017

Isomerization in the CDR2 of a monoclonal antibody: Binding analysis and factors that influence the isomerization

Lawrence W Dick1, Difei Qiu, Rosie B Wong

  • 1Analytical and Formulation Development Department, Medarex, Inc., Bloomsbury, New Jersey 08804, USA.

Biotechnology and Bioengineering
|October 7, 2009
PubMed

Isomerization of a monoclonal antibody is one of the common routes of protein degradation. An isomerization in the complementarity-determining region (CDR) was found previously and is investigated in depth in this work. Affinity analysis proves that the antibody with one isomerized heavy chain has lower binding. Binding constants were determined, and exhibited a slower on-rate in conjunction with a faster off-rate for this isomerization. To determine the role of the buffer on the rate of isomerization, this antibody was incubated in various matrices and the amount of isomerized antibody was determined by hydrophobic interaction chromatography (HIC). The rate was found to be dependent on the pH as well as the net negative charge of the buffer components that can act as proton acceptors. An Arrhenius plot was performed to predict the levels of isomerization and a comparison of real samples proved the model was correct. This work affirms that isomerization in the CDR of a therapeutic antibody is important to monitor and the formulation buffer plays a significant role in the rate of the isomerization.

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