由人免疫球蛋白转基因小鼠制备人抗乙酰胆碱受体单克隆抗体

Preparation of human monoclonal antibodies against acetylcholine receptor in human immunoglobulin loci transgenic mice

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.