An optimized approach to the rapid assessment and detection of sequence variants in recombinant protein products

Analytical and Bioanalytical Chemistry
Lowell J BradyAlain Balland

Abstract

The development of sensitive techniques to detect sequence variants (SVs), which naturally arise due to DNA mutations and errors in transcription/translation (amino acid misincorporations), has resulted in increased attention to their potential presence in protein-based biologic drugs in recent years. Often, these SVs may be below 0.1%, adding challenges for consistent and accurate detection. Furthermore, the presence of false-positive (FP) signals, a hallmark of SV analysis, requires time-consuming analyst inspection of the data to sort true from erroneous signal. Consequently, gaps in information about the prevalence, type, and impact of SVs in marketed and in-development products are significant. Here, we report the results of a simple, straightforward, and sensitive approach to sequence variant analysis. This strategy employs mixing of two samples of an antibody or protein with the same amino acid sequence in a dilution series followed by subsequent sequence variant analysis. Using automated peptide map analysis software, a quantitative assessment of the levels of SVs in each sample can be made based on the signal derived from the mass spectrometric data. We used this strategy to rapidly detect differences in sequence varia...Continue Reading

References

Apr 7, 2009·Regulatory Toxicology and Pharmacology : RTP·Rafael PonceDan Wierda
Sep 19, 2009·Nature Reviews. Genetics·D Allan Drummond, Claus O Wilke
May 28, 2010·Biotechnology and Bioengineering·Anurag KhetanThomas Ryll
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Oct 31, 2012·Biotechnology and Bioengineering·Lauren FeeneyMichael W Laird
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Citations

Dec 30, 2017·Expert Opinion on Biological Therapy·William C LamannaMartin Schiestl
Oct 27, 2018·MAbs·Douglas S RehderMatthew R Schenauer
Apr 4, 2017·Biotechnology and Bioengineering·Tzihsuan Jennifer LinKarin Anderson
Jun 26, 2021·Scientific Reports·Anushikha ThakurRamakrishnan Melarkode Subbaraman

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