A SOLID-PHASE ENZYME-LINKED IMMUNOSPOT (ELISPOT) ASSAY FOR ENUMERATION OF SPECIFIC ANTIBODY-SECRETING CELLS
A SOLID-PHASE ENZYME-LINKED IMMUNOSPOT (ELISPOT) ASSAY FOR ENUMERATION OF SPECIFIC ANTIBODY-SECRETING CELLS
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DOI：
10.1016/0022-1759(83)90308-3
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发表时间：
1983-01-01
影响因子：
2.2
通讯作者：
TARKOWSKI, A
中科院分区：
文献类型：
--
作者：
CZERKINSKY, CC;NILSSON, LA;TARKOWSKI, A
描述了固相酶连接的免疫吸附测定法（ELISPOT），用于枚举分泌特定抗体的细胞。来自免疫小鼠的脾细胞在抗原涂层的聚苯乙烯板中孵育。去除细胞后，通过免疫酶程序证明了结合的抗体，其中在琼脂糖中进行了酶 - 基底反应。黑褐色的圆形区域（斑点）位于发生抗体产生的盘子区域，并用肉眼列出。可以通过将凝胶取代液体缓冲液来对酶结合活性进行分光光度估计，从而准确地估算了分泌抗体的总量。该新测定的多功能，灵敏且易于执行，为常规的斑块形成细胞分析提供了有用的替代方法。
A solid-phase enzyme-linked immunosorbent assay (ELISPOT) is described for enumeration of cells secreting specific antibody. Spleen cells from immunized mice are incubated in antigen-coated polystyrene plates. After removal of the cells, bound antibodies are demonstrated by means of an immunoenzyme procedure in which enzyme-substrate reactions are performed in agarose. Dark-brown circular zones (spots), localized in areas of the dish where antibody production has occurred, are enumerated with the naked eye. Spectrophotometric estimation of enzyme-bound activity may be performed by substituting the gel for a liquid buffer, allowing accurate estimation of the total amount of secreted antibody. Versatile, sensitive and easy to perform, this new assay provides a useful alternative to conventional plaque-forming cell assays.