Epstein S L, Lunney J K
J Immunol Methods. 1985 Jan 21;76(1):63-72. doi: 10.1016/0022-1759(85)90481-8.
An ELISA using plates coated with mouse spleen cells has been developed for analysis of antibodies to cell surface antigens. Such assays have been used extensively with human cells or with tumor cells in various species, but application to normal mouse lymphocytes has been limited. Use of normal spleen cells allows access to the genetic resources offered by recombinant, congenic, and mutant mouse strains, in the preparation of cell-coated ELISA plates, the use of glutaraldehyde was found to be unnecessary and it was eliminated, thereby avoiding the destruction of some cell surface determinants. Poly-L-lysine, which was used to treat plates, was found to provide sufficient adherence and preservation of the cells. Binding of biotinylated monoclonal antibodies to cells could be detected at approximately 10 ng/well. In inhibition assays, unlabeled antibodies could be detected at approximately 10 ng/well. Cell-coated plates are stable once prepared, and can be stored for months before use. The assay described can be used to quantitate levels of antibody to a particular epitope, and can also be adapted for screening of fusions for monoclonal antibodies to cell surface antigens.
一种使用包被有小鼠脾细胞的酶联免疫吸附测定（ELISA）已被开发用于分析针对细胞表面抗原的抗体。此类测定已广泛应用于人类细胞或各种物种的肿瘤细胞，但在正常小鼠淋巴细胞中的应用有限。使用正常脾细胞能够利用重组、同源和突变小鼠品系所提供的遗传资源。在制备细胞包被的ELISA板时，发现无需使用戊二醛，因此将其省去，从而避免了某些细胞表面决定簇的破坏。发现用于处理板的聚-L-赖氨酸能够提供足够的细胞黏附力并保存细胞。生物素化单克隆抗体与细胞的结合在约10 ng/孔时即可检测到。在抑制测定中，未标记的抗体在约10 ng/孔时可被检测到。细胞包被的板一旦制备好就很稳定，可在使用前保存数月。所描述的测定可用于定量针对特定表位的抗体水平，也可适用于筛选针对细胞表面抗原的单克隆抗体的融合体。
