在酶联免疫吸附测定（ELISA）形式中，使用固定化α-1-抗胰蛋白酶或大豆胰蛋白酶抑制剂（SBTI）对基于细胞的疗法中的残留胰蛋白酶进行定量分析。

Quantitation of residual trypsin in cell-based therapeutics using immobilized α-1-antitrypsin or SBTI in an ELISA format.

作者信息

Braatz James A, Elias Christopher, Finny Joseph G, Tran Huan, McCaman Michael

机构信息

BioAssay Services Department, Lonza Walkersville, Inc., Walkersville, MD 21793, United States.

出版信息

J Immunol Methods. 2015 Feb;417:131-133. doi: 10.1016/j.jim.2014.12.009. Epub 2014 Dec 17.

DOI:10.1016/j.jim.2014.12.009

PMID:25527344

Abstract

An Enzyme-Linked Immunosorbent Assay (ELISA) has been developed for the quantitation of porcine trypsin as a process residual in cell therapy products based on its capture by either of two immobilized anti-trypsins, α-1-antitrypsin (α1AT) or soybean trypsin inhibitor (SBTI) followed by detection with a polyclonal goat anti-porcine trypsin-IgG conjugated with peroxidase. It was demonstrated that an extended range of antigen quantitation could be achieved that covered nearly three orders of magnitude of trypsin concentration. The utility of the assay was demonstrated by its application to samples generated in a cell-based therapeutic manufacturing setting.

摘要

已经开发出一种酶联免疫吸附测定（ELISA）方法，用于定量检测细胞治疗产品中的猪胰蛋白酶残留量。该方法基于两种固定化抗胰蛋白酶（α-1抗胰蛋白酶（α1AT）或大豆胰蛋白酶抑制剂（SBTI））之一捕获猪胰蛋白酶，然后用与过氧化物酶偶联的山羊抗猪胰蛋白酶多克隆IgG进行检测。结果表明，该方法可以实现较宽范围的抗原定量，涵盖了近三个数量级的胰蛋白酶浓度。通过将该测定方法应用于基于细胞的治疗性生产过程中产生的样品，证明了其实用性。