■ ESI-MS for the measurement of bovine and porcine somatotropins

N. Rochut, B. Bizec, F. Monteau, F. André
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引用次数: 15

Abstract

Somatotropins are proteins naturally produced by the pituitary gland in cattle. Recombinant somatotropins, which differ from their native form by several amino acids, have been synthesised in order to increase milk production in dairy cows. As their use is forbidden in Europe, control has to be performed. Until now, radioimmunoassay methods were generally used for quantification, but none is able to distinguish endogenous from recombinant somatotropins. Herein, a sensitive method based on liquid chromatography‐mass spectrometry (LC-MS) and electrospray ionisation (ESI) is presented in view to identify and quantify somatotropins in milk and plasma. Accuracy of the technique should allow the molecular mass differentiation between pituitary and recombinant somatotropins. The detection limit was measured at the nanogram level. As a second approach an enzymatic digestion was performed on recombinant somatotropin. A N-terminal digest proved to be specific and would allow to differentiate recombinant and endogenous proteins. Separation and detection of peptide fragments by ESI-LC-MS appear to be promising.
ESI-MS用于测定牛和猪的生长激素
生长激素是由牛脑垂体自然产生的蛋白质。为了增加奶牛的产奶量,已经合成了重组生长激素,它与天然形式有几个氨基酸的不同。由于它们在欧洲被禁止使用,因此必须进行控制。到目前为止,放射免疫分析法通常用于定量,但没有一种方法能够区分内源性和重组的生长激素。本文提出了一种基于液相色谱-质谱(LC-MS)和电喷雾电离(ESI)的灵敏方法,用于鉴定和定量牛奶和血浆中的生长激素。该技术的准确性应允许垂体和重组生长激素之间的分子质量区分。检测限为纳克级。第二种方法是对重组生长激素进行酶消化。n端消化被证明是特异性的,可以区分重组蛋白和内源性蛋白。用ESI-LC-MS分离和检测肽片段是很有前途的。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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