title: A new approach to quantification of mAb aggregates using peptide affinity probes.
abstract: Using mAbs as therapeutic molecules is complicated by the propensity of mAbs to aggregate at elevated concentrations, which can lead to a variety of adverse events in treatment. Here, we describe a proof-of-concept for new methodology to detect and quantify mAb aggregation. Assay development included using an aggregated mAb as bait for screening of phage display peptide library and identifying those peptides with random sequence which can recognize mAb aggregates. Once identified, the selected peptides can be used for developing quantitative methods to assess mAb aggregation. Results indicate that a peptide binding method coupled with mass spectrometric detection of bound peptide can quantify mAb aggregation and potentially be useful for monitoring aggregation propensity of therapeutic protein candidates.
authors.affiliation: Biomolecular Measurement Division, National Institute of Standards and Technology, Gaithersburg, MD 20899, USA.|Institute for Bioscience and Biotechnology Research, Rockville, MD 20850, USA.
authors.forename: Crystal S F
authors.initials: CS
authors.lastname: Cheung
authors.affiliation: Biomolecular Measurement Division, National Institute of Standards and Technology, Gaithersburg, MD 20899, USA.|Institute for Bioscience and Biotechnology Research, Rockville, MD 20850, USA.
authors.forename: Kyle W
authors.initials: KW
authors.lastname: Anderson
authors.affiliation: Institute for Bioscience and Biotechnology Research, Rockville, MD 20850, USA.
authors.forename: Pooja M
authors.initials: PM
authors.lastname: Patel
authors.affiliation: Institute for Bioscience and Biotechnology Research, Rockville, MD 20850, USA.
authors.forename: Keale L
authors.initials: KL
authors.lastname: Cade
authors.affiliation: Biomolecular Measurement Division, National Institute of Standards and Technology, Gaithersburg, MD 20899, USA.
authors.forename: Karen W
authors.initials: KW
authors.lastname: Phinney
authors.affiliation: Biomolecular Measurement Division, National Institute of Standards and Technology, Gaithersburg, MD 20899, USA.|Institute for Bioscience and Biotechnology Research, Rockville, MD 20850, USA.
authors.forename: Illarion V
authors.initials: IV
authors.lastname: Turko
mesh_terms: D000911:Antibodies, Monoclonal / Q000737:chemistry* / Q000378:metabolism; D002853:Chromatography, Liquid; D004797:Enzyme-Linked Immunosorbent Assay; D005455:Fluorescent Antibody Technique; D006801:Humans; D015335:Molecular Probes; D010455:Peptides / Q000737:chemistry / Q000378:metabolism; D066329:Protein Aggregates; D011485:Protein Binding; D053719:Tandem Mass Spectrometry
chemical_list: D000911:Antibodies, Monoclonal; D015335:Molecular Probes; D010455:Peptides; D066329:Protein Aggregates