水蛭素及重组水蛭素体内外分析方法的研究进展

Development in analytical methods for hirudin and recombinant hirudin

水蛭素是天然的凝血酶抑制剂,由65个氨基酸组成的多肽,相对分子质量为7 000,等电点为3.8。大量的研究表明,水蛭素具有强大的抗血栓和抗凝血作用。随着基因工程技术的发展,重组水蛭素产品也得到了开发,从而克服了天然水蛭素产量的不足。重组水蛭素与天然水蛭素具有相似的结构和相同的药理活性。本文综述了近20年来国内外关于水蛭素及重组水蛭素的体内外分析方法,主要有以下4个方面:①生物测定法,包括凝血酶滴定法、抗凝法、生色底物法、蝰蛇毒凝血时间测定法。②免疫测定法,包括放射免疫测定法、酶联免疫吸附分析法、三抗体夹心酶免疫吸附法。③同位素标记法。④光散射法等。本文旨在为重组水蛭素的进一步研究和质量控制提供参考。

The natural hirudin is the most effective anticoagulant.It is a single polypeptide chain of 65 amino acids with higher contents of glutamine and asparagine,which has a relative molecular weight of 7 000 and isoelectric point（PI） of 3.8.Lots of research show that hirudin has stronger antithrombotic and anticoagulation effects.With the development of genetic engineering technology,mass exploitation of recombinant hirudin（rH） make up for avoiding the deficiency of natural hirudin.The rH and natural hirudin have the similar structure and the same pharmacological activities.To provide reference for research and quality control of rH,this review summarized all kinds of in vitro and in vivo methods for analyzing rH and natural hirudin at home and abroad in the past 20 years.There are four kinds as following： ① bioassay methods including titration with thrombin,anticoagulation,chromogenic substrates and ecarin clotting time.② immunoassays including radioimmunoassay,ELISA and triple antibodies sandwich enzyme-linked immunoadsorbent assay.③ isotope label method.④ light scattering method.This article is aim to proide reference for fmther study rH omd its quality control.