Detection of Isopeptide Bonds in Monoclonal Antibody Aggregates.

ABSTRACT:

Purpose

A major difficulty in monoclonal antibody (mAb) therapeutic development is product aggregation. In this study, intermolecular isopeptide bonds in mAb aggregates were characterized for the first time. We aim to propose a mechanism of covalent aggregation in a model antibody using stressed studies at raised temperatures to aid in the understanding of mAb aggregation pathways.

Methods

Aggregate fractions were generated using raised temperature and were purified using size-exclusion chromatography (SEC). The fractions were tryptically digested and characterized using liquid chromatography hyphenated to tandem mass-spectrometry (LC-MS/MS).

Results

An increased amount of clipping between aspartic acid and proline in a solvent accessible loop in the constant heavy 2

SUBMITTER: Powell T 

PROVIDER: S-EPMC8497302 | biostudies-literature | 2021 Sep

REPOSITORIES: biostudies-literature