碱性磷酸酶-抗碱性磷酸酶斑点酶联免疫吸附试验方法的建立

ESTABLISHMENT OF APAAP DOT-ELISA METHOD

本文建立碱性磷酸酶-抗碱性磷酸酶(APAAP)斑点酶联免疫吸附试验(ELISA)方法,对实验条件加以选择,同时检测了鼠源单克隆抗体(McAb)及甲胎蛋白(AFP)及免疫球蛋白G(IgG).结果表明:该法所选用的3种包被液,pH9.5 0.05 mol/L碳酸缓冲液,pH7.2 TBS,pH7.6 PBS无差异。底物溶液以坚固红溶液较理想。6种鼠源McAh腹水稀释8000～32000倍仍能测定。另外,可检测AFP,IgG最小量分别为10.94μg/L和152.03μg/L.APAAP-斑点ELISA可望用于McAb筛选和微量抗原的检测。

A dot-ELIA using alkaline phosphatase-anti-alkaline phosphatese technique (A-PAAP) is reported. With the peimal experiment condition determined, several kinds ofmouse monoclonal antibodies (McAb), alpha fetoprotein AFP) and human immunoglobu-lin G (IgG) were detected. The results showed that there were no difference among the coating solutions we used in the experiment and that Fast Red B salt was a suitable sub-strate solution. This method ould detect 6 kinds of mouse McAbs that were diluted asmuch as 8000 to 32000 times thinner. The smallest etectable amounts of AFP and IgG-were 10.94μg/L and 152.03μg/L respectively. The APAAP dot-LISA ethod may wellbe adopted for cloning McAbs and detecting minute aneigens.