抗兔α-2-巨球蛋白单克隆抗体及其在灵敏酶联免疫吸附测定中的应用。

Monoclonal antibodies to rabbit alpha-2-macroglobulin and their use in a sensitive ELISA assay.

作者信息

Kurdowska Anna, Miller Edmund J, Krupa Agnieszka, Noble James M, Sakao Yukinori

机构信息

Department of Biochemistry, University of Texas Health Center at Tyler, 11937 US Highway 271, Tyler, TX 75708-3154, USA.

出版信息

J Immunol Methods. 2002 Dec 15;270(2):147-53. doi: 10.1016/s0022-1759(02)00278-8.

DOI:10.1016/s0022-1759(02)00278-8

PMID:12379320

Abstract

alpha-2-macroglobulin (alpha-2-M), a plasma proteinase inhibitor, plays an important role in the pathogenesis of lung inflammation. The purpose of this study was to develop a sensitive ELISA assay for rabbit alpha-2-M to allow us to define the role of this protein in a rabbit model of lung inflammation/injury. Therefore, we developed hybridomas which secrete monoclonal antibodies (mAbs) against rabbit alpha-2-M. From the antibodies produced, two (5B6 and 3C5) were selected. Both of them were of the IgG1 subclass. 5B6 reacted with native alpha-2-M as well as with the "fast" form of alpha-2-M (native alpha-2-M or the "slow" form is converted to the "fast" form by reaction with a proteinase). On the other hand, 3C5 reacted only with "fast" form of alpha-2-M. With these antibodies, we developed two ELISA assays which were used to determine the concentration of alpha-2-M in the lung fluids from rabbits with lung injury.

摘要

α-2-巨球蛋白（α-2-M）是一种血浆蛋白酶抑制剂，在肺部炎症的发病机制中起重要作用。本研究的目的是开发一种针对兔α-2-M的灵敏酶联免疫吸附测定（ELISA）方法，以便我们确定该蛋白在兔肺部炎症/损伤模型中的作用。因此，我们制备了分泌抗兔α-2-M单克隆抗体（mAb）的杂交瘤。从产生的抗体中，挑选出两种（5B6和3C5）。它们均为IgG1亚类。5B6与天然α-2-M以及α-2-M的“快速”形式发生反应（天然α-2-M或“慢速”形式通过与蛋白酶反应转化为“快速”形式）。另一方面，3C5仅与α-2-M的“快速”形式发生反应。利用这些抗体，我们开发了两种ELISA方法，用于测定肺损伤兔肺液中α-2-M的浓度。