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从小鼠肝脏中一步法免疫吸附制备F蛋白并保留同种异体抗原位点,以及测定肝脏和血清中的浓度。

Single-step immunosorbent preparation of F-protein from mouse liver with conservation of the allo-antigenic site, and determination of concentration in liver and serum.

作者信息

Wedderburn L, Lukic M L, Edwards S, Kahan M C, Nardi N, Mitchison N A

出版信息

Mol Immunol. 1984 Oct;21(10):979-84. doi: 10.1016/0161-5890(84)90156-1.

Abstract

A single-step immunopurification procedure is described for murine protein F, in which the T-cell-defined allo-antigenic site on the protein is fully conserved. The procedure is based on the use of a newly developed monoclonal antibody. The protein is isolated as a 42,500 mol. wt (F.1) and a 43,000 mol. wt (F.2) monomer. The content in liver, as estimated by radioimmune inhibition assay, is 0.083% and the yield is approximately one third. An assay of immunogenic activity in adoptive transfer, which detects the T-cell-defined site, provides a similar estimate of content in liver. The adoptive transfer assay yields concns of F-protein in serum of young mice of 0.5-1.2 X 10(-9)M, the lowest concn of protein known to induce complete immunological tolerance.

摘要

本文描述了一种针对小鼠蛋白F的单步免疫纯化程序,该蛋白上由T细胞定义的同种异体抗原位点是完全保守的。该程序基于一种新开发的单克隆抗体的使用。该蛋白以42,500摩尔分子量(F.1)和43,000摩尔分子量(F.2)的单体形式分离出来。通过放射免疫抑制测定法估计,肝脏中的含量为0.083%,产率约为三分之一。在过继转移中检测由T细胞定义的位点的免疫原性活性测定,对肝脏中的含量提供了类似的估计。过继转移测定法得出幼鼠血清中F蛋白的浓度为0.5 - 1.2×10(-9)M,这是已知能诱导完全免疫耐受的最低蛋白浓度。

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