Klinman Dennis M, Nutman Thomas B
Center for Biologics Evaluation & Research, Food and Drug Administration, Bethesda, Maryland.
National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, Maryland.
Curr Protoc Immunol. 2001 May;Chapter 6:6.19.1-6.19.8. doi: 10.1002/0471142735.im0619s10.
The filter immunoplaque assay, otherwise called the enzyme-linked immunospot assay (ELISPOT), was initially developed to detect and quantitate individual antibody-secreting B cells. Recent modifications have improved the sensitivity of the ELISPOT assay such that cells producing as few as 100 molecules of specific protein per second can be detected. The ELISPOT assay utilizes two high-affinity cytokine-specific antibodies directed against different epitopes on the same cytokine molecule: either two monoclonal antibodies or a combination of one monoclonal antibody and one polyvalent antiserum. ELISPOT generates spots based on a colorimetric reaction that detects the cytokine secreted by a single cell. The spot represents a "footprint" of the original cytokine-producing cell. Spots are permanent and can be quantitated visually, microscopically, or electronically.
滤膜免疫斑点测定法，又称酶联免疫斑点测定法（ELISPOT），最初是为检测和定量单个分泌抗体的B细胞而开发的。最近的改进提高了ELISPOT测定法的灵敏度，以至于每秒产生低至100个特定蛋白质分子的细胞都能被检测到。ELISPOT测定法利用两种针对同一细胞因子分子上不同表位的高亲和力细胞因子特异性抗体：两种单克隆抗体或一种单克隆抗体与一种多价抗血清的组合。ELISPOT基于检测单个细胞分泌的细胞因子的比色反应产生斑点。该斑点代表原始产生细胞因子的细胞的“印记”。斑点是永久性的，可以通过目视、显微镜或电子方式进行定量。
