Iazova A K, Gusev A I, Lezhneva O M
Biull Eksp Biol Med. 1982 Jul;94(7):62-4.
Splenocytes of rats immunized with mouse alpha-fetoprotein (MAFP) and cells of mouse myeloma NS-I were used to obtain hybridomas that synthesize antibodies to MAFP (anti-MAFP). Antibody activity was determined by the two tests: solid-phase ELISA and indirect immunoperoxidase staining of the regenerating liver sections of mice poisoned with CCl4. After recloning of one of the hybridomas a clone that effectively synthesized anti-MAFP was selected and reproduced. Anti-MAFP were isolated from cultural fluid of the clone by sorption-elution on CnBr-activated sepharose 4B conjugated with purified MAFP. Monoclonal origin of anti-MAFP has been demonstrated by electrophoretic homogeneity of the preparation, and by its appurtenance to one of the subclasses of rat IgG. Anti-MAFP does not precipitate MAFP but distinctly inhibits the formation of the precipitation line by MAFP and rabbit antibodies to MAFP in the zone of diffusion of monoclonal antibodies. Monoclonal anti-MAFP were high-effective in the reaction with MAFP in both tests. Anti-MAFP specificity is directed to one of the MAFP determinants that is common to rat and human alpha-fetoprotein.
用小鼠甲胎蛋白(MAFP)免疫的大鼠脾细胞和小鼠骨髓瘤NS-1细胞用于获得合成抗MAFP抗体(抗MAFP)的杂交瘤。通过两种试验测定抗体活性:固相ELISA和对四氯化碳中毒小鼠再生肝切片的间接免疫过氧化物酶染色。在对其中一个杂交瘤进行亚克隆后,选择并繁殖出一个有效合成抗MAFP的克隆。通过在与纯化的MAFP偶联的溴化氰活化的琼脂糖4B上进行吸附洗脱,从该克隆的培养液中分离出抗MAFP。抗MAFP的单克隆来源已通过制剂的电泳均一性及其属于大鼠IgG亚类之一得到证明。抗MAFP不会使MAFP沉淀,但在单克隆抗体扩散区能明显抑制MAFP与兔抗MAFP抗体形成沉淀线。在两种试验中,单克隆抗MAFP与MAFP反应均高效。抗MAFP的特异性针对大鼠和人甲胎蛋白共有的一种MAFP决定簇。