Reimer C B, Phillips D J, Aloisio C H, Black C M, Wells T W
Division of Host Factors, Centers for Disease Control, Atlanta, Georgia 30333.
Immunol Lett. 1989 Jun 1;21(3):209-15. doi: 10.1016/0165-2478(89)90106-5.
We used an immunofluorescent sequential-saturation-of-antibody assay and an interactive computer program for Scatchard analysis to determine association constants (Ka) of 33 murine monoclonal antibodies (Mabs) specific for human IgA epitopes. Ka ranged from 0.37 to 690 x 10(7) liters per mole (an approximate 1900-fold difference). Specificity was validated with a panel of 18 highly purified IgA1 and IgA2 myeloma proteins and secretory IgA using an immunofluorometric assay. Western blots of bacterial IgA protease digests were used to locate the epitopes of IgA specific Mabs in either the Fab, Fc, or hinge region. Mabs specific for unique epitopes on secretory IgA or free secretory component (FSC) were produced and evaluated.
我们使用免疫荧光抗体序列饱和分析法和用于Scatchard分析的交互式计算机程序来确定33种针对人IgA表位的鼠单克隆抗体（Mab）的结合常数（Ka）。Ka范围为0.37至690×10⁷升/摩尔（相差约1900倍）。使用免疫荧光测定法，通过一组18种高度纯化的IgA1和IgA2骨髓瘤蛋白以及分泌型IgA验证特异性。细菌IgA蛋白酶消化产物的蛋白质印迹法用于定位IgA特异性Mab在Fab、Fc或铰链区的表位。制备并评估了针对分泌型IgA或游离分泌成分（FSC）上独特表位的Mab。
