Chen P P, O'Hair C H, Zuraw B L, Katz D H
J Immunol Methods. 1983 Mar 11;58(1-2):59-71. doi: 10.1016/0022-1759(83)90263-6.
During the last few years studies in rats and mice have demonstrated IgE-binding factors, some of which have IgE-selective regulatory activities. This prompted us to develop a rapid, sensitive screening assay for measuring IgE-binding factors in humans. The principle of the assay is to measure the degree of inhibition of binding between anti-human IgE antibodies and human IgE. Thus, 200 pg IgE plus testing samples were added to each well precoated with anti-human IgE antiserum. After an overnight incubation, the wells were washed and radiolabeled anti-IgE antibodies were added to the wells. Under the optimum conditions, the assay can detect 10(-11)M anti-human IgE antibodies. With this assay, we have been able to detect IgE-binding factors in the supernatants of 2 human B cell lines which bear Fc receptors for IgE (FcR epsilon) on their surface membranes (e.g., WIL-2 and RPMI 8866), but not in the supernatants of DAUDI cells (a human cell line without FcR epsilon). Furthermore, the IgE-binding factors of WIL-2 cells were specifically adsorbed to, and eluted from, IgE-coupled Sepharose, but not BSA-Sepharose. These findings prove that the inhibition factors are indeed human IgE-binding factors, and that the assay described herein is a specific and sensitive screening assay for detecting human IgE-binding factors.
在过去几年中，对大鼠和小鼠的研究已证实存在IgE结合因子，其中一些具有IgE选择性调节活性。这促使我们开发一种快速、灵敏的筛选检测方法来测定人类中的IgE结合因子。该检测方法的原理是测量抗人IgE抗体与人IgE之间结合的抑制程度。因此，将200 pg IgE与测试样品加入到预先包被有抗人IgE抗血清的每个孔中。过夜孵育后，洗涤孔并向孔中加入放射性标记的抗IgE抗体。在最佳条件下，该检测方法能够检测到10(-11)M的抗人IgE抗体。通过这种检测方法，我们能够在两种人B细胞系（其表面膜上带有IgE的Fc受体（FcRε），例如WIL-2和RPMI 8866）的上清液中检测到IgE结合因子，但在DAUDI细胞（一种没有FcRε的人细胞系）的上清液中未检测到。此外，WIL-2细胞的IgE结合因子特异性吸附到IgE偶联的琼脂糖上，并从其上洗脱下来，但不能从BSA琼脂糖上洗脱。这些发现证明抑制因子确实是人类IgE结合因子，并且本文所述的检测方法是一种用于检测人类IgE结合因子的特异性和灵敏的筛选检测方法。
