Kenimer J G, Kim K J, Probst P G, Manclark C R, Burstyn D G, Cowell J L
Laboratory of Cellular Physiology, Food and Drug Administration, Bethesda, MD 20892.
Hybridoma. 1989 Feb;8(1):37-51. doi: 10.1089/hyb.1989.8.37.
Six monoclonal antibodies (MAbs) to pertussis toxin (PT) have been generated and characterized. Five of these MAbs (3CX4, 3C4D, 6D11C, 6FX1, and X2X5) interact with determinants on the catalytic subunit (S1) of PT, and one (6DX3) is specific for subunit S4. The MAbs are divided into three groups based upon their ability to neutralize the effects of PT in a Chinese hamster ovary (CHO) cell assay. Three of the MAbs (3CX4, 3C4D and 6D11C) had high neutralization titers, one MAb (6FX1) displayed weak neutralizing activity, and two MAbs (X2X5 and 6DX3) had no neutralizing ability. The combination of one of the high titer MAbs (3CX4) with the low titer MAb (6FX1) resulted in a synergistic enhancement of neutralizing capability. F(ab')2 fragments prepared from MAb's 3CX4 and X2X5 displayed activities in the CHO-cell assay which were identical to the native MAb's. The ability of the MAbs to neutralize PT in the CHO-cell toxin neutralization assay correlated with their ability to inhibit the in vitro ADP-ribosylation of PT. A competition ELISA method demonstrated that this panel of MAbs recognizes at least four separate epitopes on the PT molecule. Biotin-conjugated MAbs were shown to be useful reagents to probe the interaction of pertussis toxin with fetuin.
已制备并鉴定了六种针对百日咳毒素（PT）的单克隆抗体（MAb）。其中五种MAb（3CX4、3C4D、6D11C、6FX1和X2X5）与PT催化亚基（S1）上的决定簇相互作用，一种（6DX3）对亚基S4具有特异性。根据它们在中国仓鼠卵巢（CHO）细胞试验中中和PT作用的能力，这些MAb被分为三组。三种MAb（3CX4、3C4D和6D11C）具有高中和效价，一种MAb（6FX1）表现出弱中和活性，两种MAb（X2X5和6DX3）没有中和能力。高滴度MAb之一（3CX4）与低滴度MAb（6FX1）的组合导致中和能力的协同增强。从MAb 3CX4和X2X5制备的F(ab')2片段在CHO细胞试验中显示出与天然MAb相同的活性。MAb在CHO细胞毒素中和试验中中和PT的能力与其抑制PT体外ADP核糖基化的能力相关。竞争ELISA方法表明，这组MAb识别PT分子上至少四个不同的表位。生物素偶联的MAb被证明是探测百日咳毒素与胎球蛋白相互作用的有用试剂。
