Canalias F, Visvikis A, Thioudellet C, Siest G
Centre du Médicament, URA CNRS 597 30, Nancy, France.
Clin Chem. 1994 Jul;40(7 Pt 1):1251-7.
The serum activity of human pancreatic lipase (HPL; EC 3.1.1.3), the main lipolytic enzyme secreted by the pancreas, is a valuable marker of pancreatic disorders. However, determining lipase activity in human serum is difficult because the substrates used vary in their lipase specificity. A lipase reference material is therefore needed. We describe the production of recombinant HPL (rHPL) in V79 Chinese hamster lung cells. A cDNA encoding the sequence of HPL was subcloned into the pcDNAI eukaryotic expression vector, which was then used to transfect V79 cells. The 50-kDa purified recombinant enzyme is fully active, is glycosylated, and has a pI of 7.0. The catalytic properties of rHPL, determined by using triolein as the substrate, were similar to those of the native enzyme. The V79rHPL cell line we developed might be useful for the production of rHPL suitable for the preparation of a reference material.
人胰脂肪酶（HPL；EC 3.1.1.3）是胰腺分泌的主要脂解酶，其血清活性是胰腺疾病的重要标志物。然而，由于所用底物的脂肪酶特异性不同，测定人血清中的脂肪酶活性较为困难。因此，需要一种脂肪酶参考物质。我们描述了在V79中国仓鼠肺细胞中生产重组HPL（rHPL）的方法。将编码HPL序列的cDNA亚克隆到pcDNAI真核表达载体中，然后用于转染V79细胞。纯化后的50 kDa重组酶具有完全活性，经过糖基化修饰，其等电点为7.0。以三油酸甘油酯为底物测定的rHPL催化特性与天然酶相似。我们开发的V79rHPL细胞系可能有助于生产适合制备参考物质的rHPL。
