Hancock R J, Yendle J E, Bradley B A
U.K. Transplant Service, Bristol, England.
Tissue Antigens. 1989 Apr;33(4):437-44. doi: 10.1111/j.1399-0039.1989.tb01692.x.
Both monoclonal human antibodies to HLA-DR antigens and supernatants from oligoclonal B-cell lines can be conveniently screened for activity by microELISA assays which use 1/10 of the volume of reagents used in conventional ELISA assays. Target cells are fixed to the bases of wells in Terasaki plates, 5 microliters volumes of supernatants incubated in these wells, and target bound antibody detected by peroxidase-conjugated anti-immunoglobulin followed by the substrate ABTS(2,2'-azino-di-(3-ethylbenzthiazoline sulphonate). The plates are read on a micro EIA reader. Supernatants can also be assayed for immunoglobulin content and isotype in Terasaki plates by coating the wells with isotype-specific anti-immunoglobulin, adding test supernatant and developing with appropriate peroxidase-conjugated anti-immunoglobulin sera and ABTS. When assaying for immunoglobulin content, the plates can be read either with a reader or by eye. Advantages and modifications of these procedures are discussed. There are no apparent practically important disadvantages to these procedures as compared with more conventional ELISA assays.
针对HLA - DR抗原的单克隆人抗体和寡克隆B细胞系的上清液，均可通过微酶联免疫吸附测定（microELISA）方便地筛选活性，该方法使用的试剂量仅为传统酶联免疫吸附测定的1/10。将靶细胞固定在特拉斯基板（Terasaki plates）孔的底部，在这些孔中加入5微升体积的上清液，通过过氧化物酶偶联的抗免疫球蛋白检测与靶标结合的抗体，随后加入底物ABTS（2,2'-叠氮基-二-（3-乙基苯并噻唑啉磺酸盐）。使用微型酶免疫分析仪读取平板。上清液也可在特拉斯基板中检测免疫球蛋白含量和同种型，方法是用同种型特异性抗免疫球蛋白包被孔，加入测试上清液，并用适当的过氧化物酶偶联抗免疫球蛋白血清和ABTS显色。检测免疫球蛋白含量时，平板既可用分析仪读取，也可肉眼观察。讨论了这些方法的优点和改进之处。与更传统的酶联免疫吸附测定相比，这些方法没有明显的实际重要缺点。
