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应用两种针对不同抗原位点的单克隆抗体(MeAb)建立的双位点一步法ELISA,简称两点一步法,检测抗原物质,不仅操作简便、快速,而且还可以达到改善检测系统灵敏度、精确性及重复性的目的。 1981年Votila首先报道用二点一步法检测人血清甲胎蛋白(AFP)含量,证明了这一方法的优越性。1986年刘杲等报道了抗人AFP McAb的制备、双位点夹心ELISA的建立及在检测血清AFP中的初步应用。1987年周鸣、刘杲报道了检测血清AFP的两点一步法。我们应用两种抗AFP不同抗原位点的McAb,通过选择合适的抗体提纯方法,应用马来酰亚胺法制备辣根过氧化物酶(HRP)-Fab’结合物以及选用灵敏的四甲基联苯胺(TMB)底物等方面的工作,建立了适合于临床检测用的血清AFP两点一步ELISA测定法。
Two-site one-step ELISA using two monoclonal antibodies against different antigenic sites (MeAbs), referred to as two-point one-step method, to detect antigenic substances is not only easy to operate and fast, but also can improve the sensitivity and accuracy of the detection system Sexual and repetitive purposes. Votila first reported in 1981 using a two-point method to detect human serum alpha-fetoprotein (AFP) content, proved the superiority of this method. In 1986, Liu et al. Reported the preparation of anti-human AFP McAb, the establishment of double-site sandwich ELISA and its preliminary application in the detection of serum AFP. Zhou Ming in 1987, Liu Ying reported detection of serum AFP two one-step method. We used two McAbs against different antigenic sites of AFP by selecting the appropriate antibody purification method, using horse-imide method to prepare horseradish peroxidase (HRP) -Fab ’conjugate and using sensitive tetramethyl Benzidine (TMB) substrate and other aspects of the work, the establishment of a suitable clinical test serum AFP two-point ELISA assay.