目的:从人免疫球蛋白(Ig)转基因小鼠制备人源性抗乙酰胆碱受体(AChR)单克隆抗体。方法:以电鳐(Torpedo)AChR(tAChR)作为基础免疫原,分别以tAChR或人AChR(hAChR)α亚单位1~210位氨基酸(Hα1-210)与thioredoxin(Trx)融合制备的融合蛋白Trx-Hα1-210作为加强免疫原,注射人Ig转基因小鼠。应用ELISA检查由该小鼠制备的杂交瘤细胞培养上清液中人源性抗tAChR或抗Trz-Hα1-210单克隆抗体的分泌,应用放射免疫测定法(RIA)测定抗tAChR或抗Trx-Hα1-210单克隆抗体与hAChR的交叉反应性。结果:从tAChR作为加强免疫原组小鼠得到433株分泌人源性抗tAChR单克隆抗体的细胞株,从抗Trx-Hα1-210作为加强免疫原组小鼠得到20株分泌人源性抗Trx-Hα1-210单克隆抗体的细胞株。但这些人源性抗体均不能与hAChR起交叉反应。结论:由人Ig转基因小鼠制备人源性抗AChR单克隆抗体是可行的。
Objective :To prepare human monoclonal antibodies (mAbs) against acetylcholine receptor(AChR) in human immunoglabldin (Ig) loci transgenic mice. Methods:Human Ig loci transgenic mice were immunized with Torpedo AChR(tAChR) as primary immunization, and with tAChR or Trx-Hαl-210, a fusion protein of thioredoxin(Trx) with amino acids 1-210 in α subunit of human AChR( hAChR), as boost immunization respectively. Hybridomas were made from the mice, and mAbs selected for the specificities of anfi-tAChR or anfi-Trx-Hαl-210 using ELISA. Cross-reactivity of the mAbs with hAChR was determined by radioimmunoassay (RIA). Results:433 strains of hybfidomas, which secreted anti-tAChR mAbs, were obtained from the mice immunized with tAChR as boost immunization, and 20 strains of hybfidomas, which secreted anti-Trx-Hαl-210 mAbs, were obtained from the mice immunized with Trx-Hαl-210 as boost immunization. However, none of the anti-tAChR mAbs or anti-Trx-Hαl-210 mAbs was able to cross-react with hAChR in RIA. Conclusion:It is possible to prepare human anti-AChR mAbs from human Ig loci transgenic mice.