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Multiplexed on-column protein digestion and capillary electrophoresis for high-throughput comprehensive peptide mapping.

作者信息

He Yan, Zhong Wenwan, Yeung Edward S

机构信息

Ames Laboratory, US Department of Energy, Iowa 50011, USA.

出版信息

J Chromatogr B Analyt Technol Biomed Life Sci. 2002 Dec 25;782(1-2):331-41. doi: 10.1016/s1570-0232(02)00698-0.

DOI:10.1016/s1570-0232(02)00698-0
PMID:12458016
Abstract

A novel scheme based on multiplexed capillary electrophoresis (CE) has been developed for high-throughput, low-cost and comprehensive peptide mapping. Orthogonal peptide maps of the protein of interest were obtained by using multiple reaction conditions with three different enzymes (trypsin, pepsin, and chymotrypsin), and multiple separation conditions with six zone electrophoresis buffers and two micellar electrokinetic chromatography (MEKC) buffers. Fifteen nanoliters of two protein samples (beta-lactoglobulin A and beta-lactoglobulin B) were separately mixed on-column and digested independently at 37 degrees C for 10 min to produce peptides in a 20-capillary system. The resulting peptides were detected simultaneously at 214 nm by a photodiode array detector. The overall analysis time from reaction to detection was about 40 min.

摘要

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