一种用于直接筛选针对过敏原分子的单克隆抗体的四步法夹心放射免疫测定。

A four-step sandwich radioimmunoassay for direct selection of monoclonal antibodies to allergen molecules.

作者信息

Ley V, Corbí A L, Sánchez-Madrid F, Carreira J C

出版信息

J Immunol Methods. 1985 Nov 28;84(1-2):265-70. doi: 10.1016/0022-1759(85)90433-8.

Abstract

A 4-step radioimmunoassay has been devised for direct identification of monoclonal antibodies (MAb) directed to IgE-binding molecules. Polyvinyl chloride wells coated with purified anti-mouse kappa chain MAb (187-1) were successively incubated with: (1) MAb-containing hybridoma supernatants, (2) allergen extract. (3) allergic patients' serum pool, and (4) 125I-labeled anti-human IgE antiserum, to detect MAb-allergen-IgE complexes. MAb to allergens from Parietaria judaica pollen and Dermatophagoides mites have been selected with this screening procedure. The affinity-purified allergen molecules completed the binding of IgE to allergen extracts coated to paper discs in a RAST inhibition assay, confirming the anti-allergen specificity of the selected MAb. This screening method is sensitive enough to allow detection of MAb directed to poorly represented allergens.

摘要

已设计出一种四步法放射免疫测定法，用于直接鉴定针对IgE结合分子的单克隆抗体（MAb）。用纯化的抗小鼠κ链单克隆抗体（187-1）包被的聚氯乙烯孔依次与以下物质孵育：（1）含单克隆抗体的杂交瘤上清液，（2）变应原提取物，（3）过敏患者血清池，以及（4）125I标记的抗人IgE抗血清，以检测单克隆抗体-变应原-IgE复合物。通过这种筛选程序已筛选出针对墙草花粉和粉尘螨变应原的单克隆抗体。在放射性变应原吸附抑制试验中，亲和纯化的变应原分子完成了IgE与包被在纸盘上的变应原提取物的结合，证实了所选单克隆抗体的抗变应原特异性。这种筛选方法灵敏度足够高，能够检测针对低表达变应原的单克隆抗体。