Journal
BIOANALYSIS
Volume 10, Issue 13, Pages 1039-1054Publisher
FUTURE SCI LTD
DOI: 10.4155/bio-2017-0282
Keywords
antibody; biotherapeutic quantitation; denatured and native protein conformation; immunocapture; intact top-down protein quantitation; multidimensional LC-MS; non-specific binding; NSB; top-down QTOF LC/MS
Funding
- Merck Research Laboratories
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Aim: The requirements for developing antibody biotherapeutics benefit from understanding the nature and relevant aspects of the entire molecule. The method presented herein employs on-line multidimensional LC-quadrupole time-of-flight (QTOF)-MS for the quantitative determination of an antibody isolated from biological samples while maintaining the intact native biologically active conformation of the antibody. Results: Following method optimization for a model antibody, an incurred biotherapeutic in cynomologus monkey was quantified in its intact top-down native conformation. A partial method validation demonstrated acceptable precision and accuracy although improved sensitivity requires further studies. Conclusion: An on-line multidimensional LC-MS approach presents a proof-of-principle example for quantifying an intact, native antibody isolated from an incurred biological sample via immunoaffinity techniques coupled with top-down QTOF LC-MS bioanalysis.
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