Micheel B, Karsten U, Fiebach H
J Immunol Methods. 1981;46(1):41-6. doi: 10.1016/0022-1759(81)90330-6.
A solid-phase immunofluorescence assay (SIFA) is described for screening of monoclonal hybridoma antibodies against soluble antigens. The test is simple, needs a small amount of material (20-100 ng of labeled antigen per sample) and allows a large number of samples (500-1 000) to be tested in 2 days. The assay is performed in microtitration plates with small cellulose nitrate discs as the solid phase. Anti-mouse IgG, the sample to be tested and FITC-labeled antigen are successively incubated with the discs. The fluorescence intensity of the surface of the discs is then measured with a microfluorometer. The assay has been used to select hybridomas against human serum albumin. Comparison of the immunofluorescence assay with a radioimmunoassay using conventional antisera and hybridoma antibodies similar sensitivity for both assays.
本文描述了一种用于筛选针对可溶性抗原的单克隆杂交瘤抗体的固相免疫荧光测定法（SIFA）。该测试简单，所需材料量少（每个样品20 - 100 ng标记抗原），并能在2天内检测大量样品（500 - 1000个）。该测定在微量滴定板中进行，以小硝酸纤维素圆盘作为固相。抗小鼠IgG、待测样品和异硫氰酸荧光素（FITC）标记的抗原依次与圆盘孵育。然后用微量荧光计测量圆盘表面的荧光强度。该测定法已用于筛选针对人血清白蛋白的杂交瘤。将免疫荧光测定法与使用常规抗血清和杂交瘤抗体的放射免疫测定法进行比较，两种测定法具有相似的灵敏度。
