A New Approach to Assess mAb Aggregation

Methods Mol Biol. 2020;2131:245-254. doi: 10.1007/978-1-0716-0389-5_12.

Abstract

A proof of concept for new methodology to detect and potentially quantify mAb aggregation is presented. Assay development included using an aggregated mAb as bait for screening of a phage display peptide library and identifying those peptides with random sequence which can recognize mAb aggregates. The selected peptides can be used for developing homogeneous quantitative methods to assess mAb aggregation. Results indicate that a peptide-binding method coupled with fluorescence polarization detection can detect mAb aggregation and potentially monitor the propensity of therapeutic protein candidates to aggregate.

Keywords: Aggregation; FITC-peptide; Monoclonal antibodies; Next-generation sequencing; Peptide phage display.

MeSH terms

  • Antibodies, Monoclonal / chemistry*
  • Antibodies, Monoclonal / genetics
  • Fluorescein-5-isothiocyanate / chemistry*
  • Fluorescence Polarization
  • High-Throughput Nucleotide Sequencing
  • Peptide Library
  • Peptides / analysis*
  • Peptides / chemistry
  • Peptides / genetics
  • Proof of Concept Study
  • Protein Aggregates

Substances

  • Antibodies, Monoclonal
  • Peptide Library
  • Peptides
  • Protein Aggregates
  • Fluorescein-5-isothiocyanate