摘要
在进行固相ELISA双夹心法时,要选择两种配对的单克隆抗体(McAb)殊非易事。本文用不同McAb的混合物与另一种McAb进行配对夹心,获得了较好的效果。实验表明,在心肌肌球蛋白轻链(CM—LC)的固相ELISA双夹心体系中,以抗CM-LCMcAb(1G6)铺底,(2B4及2F6)混合物为后续复盖抗体,最低检出量可低达10ng/mL,其检出率较单独2B4或单独2F6作为后续复盖抗体者高5—10倍。而若反之,以(2B4及2F6)混合物铺底,1G6作为后续复盖抗体,则其最低检出量竟高至200ng/mL,还不如以其中之一铺底为佳。在人绒毛膜促性腺激素(HCG)的检测体系中,用多克隆抗体与单克隆抗体配对的研究中,也获得了类似的实验结果。
A sensitive ELISA was developed by using McAbs mixture paired with a single McAb. In the assay for human cardiac myosin light chain (CM-LC), if the microtiter plate was first coated with a single antiCM-LC McAb (lG6), and a McAbs mixture (2B4 & 2F6) was used as a successive antibody, the assay had a sensitivity of 10 ng/mL. It is 5 to 10 fold sensitive than 2B4 or 2F6 alone as a successive antibody. On the contray, when the McAbs mixture (2B4 & 2Fd) was used as coated antibody and 1G6 as succesive antibody, the sensitivity was dramatically decreased to 200ng/mL. In the assay for human chorionic gonadotropin (HCG), a similar phenomena was observed when a polyclonal anti-HCG and an anti-HCG McAb were chosen as paired antibodies.
关键词
单克隆抗体
固相ELISA
Monoclonal antibody Solid phase enzyme linked immunosorbent assay
