一种用于通过亲和色谱法纯化免疫球蛋白和Fab片段的人工蛋白L。

An artificial protein L for the purification of immunoglobulins and fab fragments by affinity chromatography.

作者信息

Roque A Cecília A, Taipa M Angela, Lowe Christopher R

机构信息

Centro de Engenharia Biológica e Química, Instituto Superior Técnico, 1049-001 Lisboa, Portugal.

出版信息

J Chromatogr A. 2005 Feb 4;1064(2):157-67. doi: 10.1016/j.chroma.2004.11.102.

DOI:10.1016/j.chroma.2004.11.102

PMID:15739883

Abstract

The development and characterization of an artificial protein L (PpL) for the affinity purification of antibodies is described. Ligand 8/7, which emerged as the lead from a de novo designed combinatorial library of ligands, inhibits the interaction of PpL with IgG and Fab by competitive ELISA and shows negligible binding to Fc. The ligand 8/7 adsorbent (Ka approximately 10(4) M(-1)) compared well with PpL in binding to immunoglobulins from different classes and sources and, in addition, bound to IgG1 with K and lambda isotypes (92% and 100% of loaded protein) and polyclonal IgG from sheep, cow, goat and chicken. These properties were also reflected in the efficient isolation of immunoglobulins from crude samples.

摘要

本文描述了一种用于抗体亲和纯化的人工蛋白L（PpL）的开发和特性。配体8/7是从一个从头设计的组合配体库中筛选出的先导物，通过竞争性ELISA抑制PpL与IgG和Fab的相互作用，并且与Fc的结合可忽略不计。配体8/7吸附剂（Ka约为10(4) M(-1)）在结合不同类别和来源的免疫球蛋白方面与PpL相当，此外，它还能结合IgG1的K和λ同种型（分别为负载蛋白的92%和100%）以及来自绵羊、牛、山羊和鸡的多克隆IgG。这些特性也体现在从粗样品中高效分离免疫球蛋白上。