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通过单克隆抗体制备技术得到三株特异结合半抗原 4 ( GSH-S-DNP二苄酯 )的单克隆抗体 HB4 ,HB5和 HB7.抗体经两步化学诱变得到具有细胞谷胱甘肽过氧化物酶 ( c GPX)活性的含硒抗体酶 m HB4 ,m HB5和 m HB7,活力分别为 1 70 ,1 867,32 U/μmol.其中 m HB5的活力是天然兔肝 c GPX的 0 .32倍 ,m 4 A4的1 .51倍 .等离子体 -质谱 ( ICP/MS)测得每分子含硒抗体酶分子中大约存在 2个硒原子 .m HB5的最适 p H为8.6～ 8.8.在 p H值范围为 7.0和 37℃条件下 ,m HB5催化 GSH和 H2 O2 或 t-ROOH反应的二级速率常数为 :k+ 1 ( H2 O2 ) 9.71× 1 0 6 L /( mol· min) ,k+ 1 ( t-ROOH) 5.99× 1 0 5 L/( mol· min) . m HB5使非酶催化反应速率提高了 9.8× 1 0 6和 3.7× 1 0 5倍 .
Three monoclonal antibodies HB4, HB5 and HB7 that specifically bind to hapten 4 (GSH-S-DNP dibenzyl ester) were obtained by the monoclonal antibody preparation technique. The antibody was chemically mutated in two steps to obtain a monoclonal antibody with glutathione peroxidation The activity of selenomethionine-resistant enzyme m HB4, m HB5 and m HB7 with activity of c GPX was 170 and 1867 U / μmol, respectively, and the activity of m HB5 was 0.32 Fold, 1.51 times that of m4A4.Plasma-mass spectrometry (ICP / MS) has about 2 selenium atoms per molecule of selenium-containing abzyme molecule.The optimum pH of m HB5 is 8.6-8.8 The second order rate constants for the reaction of m HB5 with GSH and H2 O2 or t-ROOH for the p H range of 7.0 and 37 ° C are: k + 1 (H2 O2) 9.71 × 10 6 L / (mol · min) k + 1 (t-ROOH) 5.99 × 10 5 L / (mol · min). The m HB5 increased the non-enzymatic reaction rate by 9.8 × 10 6 and 3.7 × 10 5 times.