Wolf H, Gattringer C, Wick G
J Immunol Methods. 1979;25(1):77-88. doi: 10.1016/0022-1759(79)90168-6.
Data are presented comparing different methods for the fractionation and enrichment, respectively, of specific antigen binding lymphoid cells from immunized chickens. The bovine serum albumin (BSA) anti-BSA system was chosen as a model. To enrich avian antigen binding cells (ABC) from a mixture of chicken peripheral blood and spleen lymphocytes 3 different methods were used: (1) separation of cells forming rosettes with antigen-coated sheep red blood cells (SRBC) from non-rosetting cells by density centrifugation; (2) isolation of ABC by their specific adherence to antigen bound to immunoadsorptive surfaces (gelatin, plastics); (3) column affinity chromatography with antigen-coated agarose, cross-linked dextran for plastic beads. The most efficient method was column affinity chromatography with antigen-coated polyacrylamide beads which affords up to 12-fold enrichment of ABC. Both the other methods are also suitable for separation and enrichment of specific ABC but can only with difficulty be adapted for processing the large numbers of cells which would be necessary, e.g., for in vivo transfer studies.
本文呈现了分别用于免疫鸡特定抗原结合淋巴细胞分级分离和富集的不同方法的比较数据。选择牛血清白蛋白（BSA）-抗BSA系统作为模型。为了从鸡外周血和脾淋巴细胞混合物中富集禽类抗原结合细胞（ABC），使用了3种不同方法：（1）通过密度离心从非玫瑰花结形成细胞中分离与抗原包被的绵羊红细胞（SRBC）形成玫瑰花结的细胞；（2）通过ABC对结合于免疫吸附表面（明胶、塑料）的抗原的特异性黏附来分离；（3）使用抗原包被的琼脂糖、交联葡聚糖或塑料珠进行柱亲和层析。最有效的方法是使用抗原包被的聚丙烯酰胺珠进行柱亲和层析，其可使ABC富集达12倍。其他两种方法也适用于特定ABC的分离和富集，但难以适用于处理大量细胞，例如用于体内转移研究所需的细胞。
