Nash A A
J Immunol Methods. 1976;12(1-2):149-61. doi: 10.1016/0022-1759(76)90105-8.
Plastic surfaces which had been coated sucessively with IgG, Staphylococal protein A (SpA) and diluted whole antiserum specific for cell membrane antigens, were used successfully to separate rabbit lymphocyte sub-populations. This method does not require the purification of SpA or antibody, and uses only small quantities of antiserum. The specificity of the cell separation method was demonstrated using various rabbit "b" locus anti-allotype sera, and the number of cells removed by the SpA-antiallotype system correlated well with the number of Ig+ve or "B" lymphocytes detected by other methods. Optimal conditions for cell attachment could readily be determined using multi-well plastic trays, and the adherent cells rapidly quantitated. Using 125I-labelled rabbit IgG (Rb IgG) or sheep IgG (Sh IgG) it was possible to quantitate the amount of IgG coupled to plastic surfaces, or binding to SpA-coated surfaces. By using 125I-labelled (As4) Rb IgG it was also possible to estimate the amount of antibody bound to SpA. The wide application of the SpA-antibody method relative to other affinity-antibody methods is discussed.
依次用免疫球蛋白G（IgG）、葡萄球菌蛋白A（SpA）和针对细胞膜抗原的稀释全抗血清包被的塑料表面，成功用于分离兔淋巴细胞亚群。该方法不需要纯化SpA或抗体，且仅使用少量抗血清。使用各种兔“b”位点抗同种异型血清证明了细胞分离方法的特异性，并且SpA-抗同种异型系统去除的细胞数量与其他方法检测到的Ig阳性或“B”淋巴细胞数量密切相关。使用多孔塑料培养板可以很容易地确定细胞附着的最佳条件，并对贴壁细胞进行快速定量。使用125I标记的兔IgG（Rb IgG）或羊IgG（Sh IgG），可以定量与塑料表面偶联或与SpA包被表面结合的IgG量。通过使用125I标记的（As4）Rb IgG，还可以估计与SpA结合的抗体量。讨论了SpA-抗体方法相对于其他亲和抗体方法的广泛应用。
