Matsuura S, Hamano Y, Kita H, Takagaki Y
Division of Biochemistry, Iatron Laboratories Inc., Chiba.
J Biochem. 1993 Aug;114(2):273-8. doi: 10.1093/oxfordjournals.jbchem.a124166.
Seven of 20 mouse monoclonal antibodies to OA, OA8-2, OA10-8, OA22-22, OA227-11, OA296-1, OA423-3, and OA958-2, were studied as to their binding to OA in organic solvents. OA423-3 (IgG1-kappa) and OA958-2 (IgG1-kappa) in 90-100% methanol retained their binding activities with both immobilized and free antibodies. Whereas OA8-2 (IgG2a-kappa), OA10-8 (IgG1-kappa), OA22-22 (IgG2a-kappa), OA227-11 (IgG1-kappa), and OA296-1 (IgM-kappa) did not bind to OA in over 50-60% methanol. The results of a non-competitive inhibition assay for OA indicated that in a methanolic or ethanolic solution, the binding ability of immobilized OA423-3 decreased as the concentration of each alcohol increased. The concentration of OA at the midpoint between the upper and lower plateaus of the inhibition curve was 0.18 ng/ml in 0% methanol and 570 ng/ml in 100%, respectively. In 0-50% of each of acetone, diethyl ether, and benzene in methanol, the binding ability of OA423-3 remained at the level in 100% methanol. OA958-2 showed similar binding properties to OA423-3. No relationship between the subclass of the immunoglobulin and the binding activity of the antibody in organic solvents was observed. These results indicate that the OA423-3 and OA958-2 antibodies are useful for the development of a new ELISA method for OA in organic solvents.
针对20种抗OA的小鼠单克隆抗体中的7种，即OA8 - 2、OA10 - 8、OA22 - 22、OA227 - 11、OA296 - 1、OA423 - 3和OA958 - 2，研究了它们在有机溶剂中与OA的结合情况。OA423 - 3（IgG1 - κ）和OA958 - 2（IgG1 - κ）在90 - 100%甲醇中对固定化抗体和游离抗体均保留其结合活性。而OA8 - 2（IgG2a - κ）、OA10 - 8（IgG1 - κ）、OA22 - 22（IgG2a - κ）、OA227 - 11（IgG1 - κ）和OA296 - 1（IgM - κ）在甲醇含量超过50 - 60%时不与OA结合。OA的非竞争性抑制试验结果表明，在甲醇或乙醇溶液中，固定化OA423 - 3的结合能力随每种醇浓度的增加而降低。抑制曲线上下平台中点处的OA浓度在0%甲醇中为0.18 ng/ml，在100%甲醇中为570 ng/ml。在甲醇中丙酮、乙醚和苯各占0 - 50%时，OA423 - 3的结合能力保持在100%甲醇中的水平。OA958 - 2表现出与OA423 - 3相似的结合特性。未观察到免疫球蛋白亚类与抗体在有机溶剂中结合活性之间的关系。这些结果表明，OA423 - 3和OA958 - 2抗体可用于开发一种针对有机溶剂中OA的新型ELISA方法。
