Peptides releasing study of serum apolipoprotein A-I and apolipoprotein B by ID-LC-MS

Abstract

Objective To investigate the digestion kinetics of Apolipoprotein A-I and B by ID-LC-MS method for accurate quantification of proteins. Methods Methodological research. The target peptides of ApoA-I and B were determined. The ApoA-I and B from 5 human serum samples on market with levels from 0.90-2.54 g/L and 0.54-1.39 g/L separately, were measured in terms of target peptides by isotope dilution liquid chromatography mass spectrometry method. The releasing amount and rate of peptides were analyzed and plotted according to different time points. The correlation coefficient R2 was calculated among peptide releasing amount between samples. Results Most peptides reached their peaks within 4 hours. The peptides VQ, DY and VS from Apo A-I, TR and FP from Apo B were released relatively slowly. After getting to their peak stage, the ratio between TEV and SIL-TEV, AK and SIL-AK, VQ and SIL-VQ presented stable state. As for Apo A-I the correlations among peptides are high, from 0.904 to 0.999. Some peptides from Apo B show lower correlations, such as TG-SV with R2 0.543 (3 h). Conclusions Peptides from Apo A-I and Apo B present different releasing properties after trypsin digestion. Proper selection of representative peptides and enzymatic conditions can benefit accurate quantification of target proteins.(Chin J Lab Med, 2018, 41: 870-874) Key words: Apolipoprotein A-I; Apolipoproteins B; Kinetics; Chromatography, liquid; Tandem mass spectrometry; Isotope labeling