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Quantification of Proteins Using Peptide Immunoaffinity Enrichment Coupled with Mass Spectrometry
Published on: July 31, 2011
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A new approach to quantification of mAb aggregates using peptide affinity probes
Crystal S F Cheung1,2, Kyle W Anderson1,2, Pooja M Patel2
1Biomolecular Measurement Division, National Institute of Standards and Technology, Gaithersburg, MD 20899, USA.
Scientific Reports
|February 11, 2017
Summary
This study introduces a new method to detect and quantify antibody (mAb) aggregation, a common issue in therapeutic treatments. The developed peptide-binding assay can monitor protein aggregation, improving therapeutic protein candidate development.
Area of Science:
- Biochemistry
- Pharmaceutical Science
- Analytical Chemistry
Background:
- Monoclonal antibodies (mAbs) are crucial therapeutics but prone to aggregation at high concentrations.
- mAb aggregation can cause adverse events, complicating their clinical use.
- Accurate detection and quantification of mAb aggregation are essential for safe and effective therapies.
Purpose of the Study:
- To develop and validate a novel methodology for detecting and quantifying monoclonal antibody (mAb) aggregation.
- To identify specific peptides that recognize mAb aggregates using phage display technology.
- To establish a quantitative assay for assessing mAb aggregation propensity.
Main Methods:
- Screening a phage display peptide library using aggregated mAbs as bait.
- Identifying peptides with high affinity for mAb aggregates.
- Developing a quantitative assay combining peptide binding with mass spectrometric detection.
Main Results:
- Successfully identified peptides capable of recognizing and binding to mAb aggregates.
- Demonstrated that the peptide-binding assay coupled with mass spectrometry can accurately quantify mAb aggregation.
- The method shows potential for monitoring the aggregation tendency of therapeutic protein candidates.
Conclusions:
- A novel, peptide-based method for detecting and quantifying mAb aggregation has been established.
- This methodology offers a promising tool for quality control and development of therapeutic antibodies.
- The assay can aid in predicting and mitigating adverse events associated with mAb aggregation.
