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Effective purification of recombinant peptide ligands for GPCR research
Date Issued
01-01-2017
Author(s)
Ashokan, Anisha
Indian Institute of Technology, Madras
Abstract
Peptide purification from natural sources and chemical synthesis is cumbersome with various shortcomings such as low yield, high cost of production, error prone, and restricted by nature of amino acids. Though recombinant DNA technology had overcome all these setbacks for larger proteins, it is still a challenge to produce peptides that are salt free and without impurities. Our approach discussed in this chapter deals with easy and effective purification of peptides of varying sizes (up to 10 kDa), expressed as fusion proteins in bacterial system. This includes cleavage of fusion affinity tag by “PreScission protease” in volatile buffer followed by selective acetonitrile precipitation of high-molecular-weight tag in order to purify peptides in solution. This method can be used to purify peptides in large scale for various biochemical and physiological studies.
Volume
142