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本文报道一种改进的硝化纤维滤膜—ELISA法(简称滤膜—ELISA),用以鉴定杂交瘤细胞培养上清McAbIg类型、其敏感度高于传统的琼脂双扩法(20倍左右),10株杂交瘤细胞中有3株的上清经琼脂双扩法未能检测出McAbIg类型者,均由滤膜—ELISA法作出鉴定。其中McAb2H_6-E_3经双扩法检测7次均为阴性反应(包括浓缩10x～40x),而用滤膜—ELISA,上清原液(1x)即鉴定出为IgG_2b亚型。经不同浓缩倍数重复12次试验,结果一致。其余7株两种方法检测结果也完全相符。说明滤膜—ELISA法的优点弥补了琼脂双扩法敏感性不高的不足之处,本法还具有节省标准抗血清的特点。
In this paper, an improved nitrocellulose filter-ELISA (hereinafter referred to as filter-ELISA) was used to identify the type of McAb Ig of hybridoma cell culture supernatant. The sensitivity was higher than that of the conventional agar double-expansion method (about 20 times) The supernatants of 3 strains from 10 hybridoma cells failed to detect McAbIg by agar double-dwell method and were identified by filter-ELISA. Among them, McAb2H_6-E_3 detected by double-diffusion method for 7 times were negative reaction (including concentrated 10x ~ 40x), while with filter-ELISA, the supernatant liquid (1x) that is identified as IgG_2b subtype. After 12 times of different concentration multiple repeat test, the results are consistent. The remaining seven strains of two test results are also fully consistent. This shows that the advantages of the membrane-ELISA method make up for the lack of sensitivity of the dual-expansion agar. This method also has the advantage of saving the standard antiserum.