Schneider M D, Eisenbarth G S
J Immunol Methods. 1979;29(4):331-42. doi: 10.1016/0022-1759(79)90004-8.
A solid-phase [125I]Protein A radioassay for anti-cell surface antibodies is described which employs target cell monolayers cultured on fenestrated polyvinyl chloride 96-well plates ('transfer plates'). The calibrated aperture in the bottom of each well is small enough to retain fluid contents by surface tension during monolayer growth, but also permits fluid to enter the wells when transfer plates are lowered into receptacles containing washing buffer or test sera. To assay for antibodies directed against target cell surface antigens, transfer plates bearing monolayers are inserted into microculture plates with corresponding 96-well geometry, thereby simultaneously sampling 96 wells. This assay allows rapid screening of hundreds of hybird cell colonies for production of antibodies with desired tissue specificity.
本文描述了一种用于抗细胞表面抗体的固相[125I]蛋白A放射测定法,该方法采用在有孔聚氯乙烯96孔板(“转移板”)上培养的靶细胞单层。每个孔底部校准的孔径小到足以在单层生长期间通过表面张力保留液体内容物,但当转移板放入含有洗涤缓冲液或测试血清的容器中时,也允许液体进入孔中。为了检测针对靶细胞表面抗原的抗体,将带有单层的转移板插入具有相应96孔几何形状的微孔板中,从而同时对96个孔进行取样。该测定法允许快速筛选数百个杂交细胞集落,以产生具有所需组织特异性的抗体。