CAJ | 학술논문

自主多样化库(ADLib)是近年新建立的一种基于体细胞突变的生产抗体的系统,其原理为以鸡的传染性法式囊淋巴细胞瘤细胞(DT40,表达鸡IgM有膜结合与分泌两种形式)为靶细胞,利用曲古酶抑菌素(TSA)抑制DT40的去乙酰活性酶,促进组蛋白乙酰化,从而造成免疫球蛋白基因高频转换,在细胞的表面展示膜结合IgM文库,最后通过抗原标记磁珠分选法从文库中选择有抗原特异性鸡源性抗体的细胞,培养细胞获得分泌性单克隆抗体IgM.进而,通过将人IgG的重链Fc段DNA序列插入DT40细胞IgM重链区相应位置,建立了合成人Fc段鸡Fab段的嵌合人IgG(重链Fc段决定免疫球蛋白类型)的ADLib系统.ADLib系统可应用于生产诊断与治疗性抗体药物的单克隆抗体(mAb).
자주다양화고(ADLib)시근년신건립적일충기우체세포돌변적생산항체적계통,기원리위이계적전염성법식낭림파세포류세포(DT40,표체계IgM유막결합여분비량충형식)위파세포,이용곡고매억균소(TSA)억제DT40적거을선활성매,촉진조단백을선화,종이조성면역구단백기인고빈전환,재세포적표면전시막결합IgM문고,최후통과항원표기자주분선법종문고중선택유항원특이성계원성항체적세포,배양세포획득분비성단극륭항체IgM.진이,통과장인IgG적중련Fc단DNA서렬삽입DT40세포IgM중련구상응위치,건립료합성인Fc단계Fab단적감합인IgG(중련Fc단결정면역구단백류형)적ADLib계통.ADLib계통가응용우생산진단여치료성항체약물적단극륭항체(mAb).
Autonomously diversifying library is a newly established system which can produce antibody based on somatic mutation.The mechanism is targeted on DT40 (DT40 is an avian leukosis virus induced bursal lymphoma cell line derived from a Hyline SC chicken,expressing membrane bound and secretion forms of chicken IgM).Trichostatin A was used to inhibit acetyl active enzyme of DT40.This in turn promotes the histone acetylation,resulting in immunoglobulin gene frequent conversion,and the display of membrane combined IgM library on the surface of cell.Finally,antigen tag magnetic separate method was used to select chicken source cells with antigen specific antibody from the library.Subsequent cell culture was used to acquire secretory monoclonal antibody IgM.Then,by inserting the DNA sequence of human IgG Fc fragment into the corresponding region of IgM heavy chain in DT40 cells,we can establish the ADLib system which synthesizes the chimeric human IgG with chicken Fab and human Fc (Fc fragment decides the type of immunoglobin heavy chain).ADLib system can be applied to the production of diagnostic and therapeutic monoclonal antibody drugs.