Scearce R M, Oie H K, Gazdar A F, Chick W L, Eisenbarth G S
J Immunol Methods. 1981;42(1):27-33. doi: 10.1016/0022-1759(81)90221-0.
A solid-phase radioimmunoassay for detection of insulin synthesized by islet cell clones is described. This assay employs anti-insulin antibody adsorbed onto fenestrated polyvinyl chloride 96-well plates ('transfer plates'). The calibrated aperture in the bottom of each transfer plate well permits fluid to enter the wells when transfer plates are lowered into microculture wells containing insulin. With this assay it is possible to rapidly screen hundreds of islet cell cultures for insulin production. We have used this assay to facilitate cloning of the RIN rat insulinoma cell line. The assay readily detects insulin synthesis by RIN cells and [125I]insulin is not displayed by culture medium from cells which do not produce insulin. The transfer plate format should be applicable to semiautomate other radioimmunoassays.
本文描述了一种用于检测胰岛细胞克隆合成胰岛素的固相放射免疫测定法。该测定法采用吸附在有孔聚氯乙烯96孔板（“转移板”）上的抗胰岛素抗体。每个转移板孔底部校准的孔径使得当转移板放入含有胰岛素的微量培养孔中时，液体能够进入孔内。通过这种测定法，可以快速筛选数百个胰岛细胞培养物以检测胰岛素的产生。我们已使用该测定法来促进RIN大鼠胰岛素瘤细胞系的克隆。该测定法能轻松检测RIN细胞合成的胰岛素，且不产生胰岛素的细胞培养基不会显示出[125I]胰岛素。转移板形式应适用于其他放射免疫测定法的半自动操作。
