Back to Search
Start Over
Glu-C, an alternative digestive enzyme for the quantitative LC-MS/MS analysis of an IgG-based antibody biotherapeutic
- Source :
- Bioanalysis. 10(13)
- Publication Year :
- 2018
-
Abstract
- Aim: LC–MS/MS bottom-up quantitation of proteins has become increasingly popular with trypsin as the most commonly used protease. However, trypsin does not always yield suitable surrogate peptides. An alternative enzyme, Glu-C, was used to generate surrogate peptides for quantifying a bispecific IgG1 biotherapeutic antibody in preclinical matrices. Materials and methods: IgG1 was quantified by pellet digestion using an Acquity UPLC coupled with a Xevo TQ-S mass spectrometer. Results: Two generic LC–MS/MS methods (heavy and light chain) were developed which afforded acceptable precision and accuracy, and an lower limit of quantitation of 1 μg/ml in three preclinical matrices. A small nonsignificant bias was observed when cynomolgus serum LC–MS/MS results were compared with electrochemiluminescent immunoassay data. Conclusion: Glu-C was successfully used as an alternative digestion enzyme for bottom-up quantitation of an IgG1 in matrices from multiple preclinical species, with good agreement with electrochemiluminescent immunoassay data.
- Subjects :
- 0301 basic medicine
Quality Control
medicine.medical_treatment
Clinical Biochemistry
Tandem mass spectrometry
01 natural sciences
Immunoglobulin G
Analytical Chemistry
03 medical and health sciences
Mice
Antibodies monoclonal
Tandem Mass Spectrometry
Lc ms ms
medicine
Animals
General Pharmacology, Toxicology and Pharmaceutics
Protease
biology
Chemistry
010401 analytical chemistry
Serine Endopeptidases
Antibodies, Monoclonal
General Medicine
Trypsin
0104 chemical sciences
Rats
Medical Laboratory Technology
Macaca fascicularis
030104 developmental biology
Biochemistry
Digestive enzyme
Calibration
biology.protein
Antibody
medicine.drug
Chromatography, Liquid
Subjects
Details
- ISSN :
- 17576199
- Volume :
- 10
- Issue :
- 13
- Database :
- OpenAIRE
- Journal :
- Bioanalysis
- Accession number :
- edsair.doi.dedup.....ad6bc830b786877dc1d721a34b9ba09f