ArticleSe pu = Chinese journal of chromatography2022
[Nonspecific adsorption evaluation and general minimization strategy in peptide analysis based on ultra-performance liquid chromatography-mass spectrometry].
Article in Se pu = Chinese journal of chromatography, 2022. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Not yet cited in PubMed.
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Abstract
Proteomics technology is being increasingly used in the development of novel therapeutic peptides and protein drugs, and also in the intensive search for clinical biomacromolecule diagnostic biomarkers. Ultra-performance liquid chromatography-mass spectrometry (UPLC-MS) is a rapid method to analyze peptides and proteins in low abundance. However, the nonspecific adsorption properties of peptides may lead to the loss or interference of the analytes throughout the analytical process. This unique nonspecific adsorption property is the main reason for the false negative and false positive results obtained through quantification, as well as for the poor precision, accuracy, linear range, and sensitivity, all of which impose significant challenges in the development of analytical methods. Accordingly, a general strategy was established to evaluate and reduce the negative impact of the nonspecific adsorption of peptides on UPLC-MS analysis. In this study, bovine serum albumin (BSA) was used as a model protein to explore the correlation between the physicochemical properties of 50 peptides obtained by the enzymatic digestion of BSA, as well as the degree of nonspecific adsorption. First, these peptides were classified into four categories according to their response and the degree of adsorption in the pretreatment containers and LC system. Next, the factors influencing the adsorption of 12 Class Ⅱ peptides, which were highly responsive and susceptible to adsorption, were systematically studied in terms of several aspects, including: (1) time-dependent adsorption on centrifuge tubes of three kinds (Protein-LoBind polypropylene tube and two types of polypropylene tubes); (2) time-dependent adsorption on sample vials of three kinds (Protein-LoBind polypropylene vial, polypropylene vial, and glass vial); (3) carryovers on chromatographic columns with six different stationary phases (Polar C
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