Hydrolysis and amino acid composition analysis of proteins

被引:458
作者
Fountoulakis, M [1 ]
Lahm, HW [1 ]
机构
[1] F Hoffmann La Roche & Co Ltd, Div Pharma, Preclin Cent Nervous Syst Res, Gene Technol, CH-4070 Basel, Switzerland
关键词
reviews; hydrolysis; amino acids; proteins;
D O I
10.1016/S0021-9673(98)00721-3
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
Amino acid composition analysis is a classical protein analysis method, which finds a wide application in medical and food science research and is indispensable for protein quantification. It is a complex technique, comprising two steps, hydrolysis of the substrate and chromatographic separation and detection of the residues. A properly performed hydrolysis is a prerequisite of a successful analysis. The most significant developments of the technology in the last decade consist in the (i) reduction of the hydrolysis time by the use of microwave radiation energy; (ii) improvement in the sensitivity of the residue detection, the quantification of the sensitive residues and separation of the enantiomeric forms of the amino acids; (iii) application of amino acid analysis in the large-scale protein identification by database search; and (iv) gradual replacement of the original ion exchange residue separation by reversed-phase high-performance liquid chromatography. Amino acid analysis is currently facing an enormous competition in the determination of the identity of proteins and amino acid homologs by the essentially faster mass spectrometry techniques. The amino acid analysis technology needs further simplification and automation of the hydrolysis, chromatography and detection steps to withstand the pressure exerted by the other technologies. (C) 1998 Published by Elsevier Science B.V. All rights reserved.
引用
收藏
页码:109 / 134
页数:26
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