中华微生物学和免疫学杂志
中華微生物學和免疫學雜誌
중화미생물학화면역학잡지
CHINESE JOURNAL OF MICROBIOLOGY AND IMMUNOLOGY
2012年
10期
845-849
,共5页
陈家琪%瞿爱东%祝婧烨%黄海武
陳傢琪%瞿愛東%祝婧燁%黃海武
진가기%구애동%축청엽%황해무
人IgE%单克隆抗体%抗原表位
人IgE%單剋隆抗體%抗原錶位
인IgE%단극륭항체%항원표위
Human IgE%Monoclonal antibody%Antigenic epitope
目的 利用基因重组抗原免疫小鼠,制备抗人IgE单克隆抗体,研究其特异性和功能.方法 用表达的重组人IgE-Fc免疫BLAB/c小鼠,取其脾细胞与Sp2/0细胞融合,以间接ELISA法筛选杂交瘤上清.用ELISA阻断分析鉴定其生物学功能;通过丙氨酸突变扫描分析确定其抗原表位.结果 利用杂交瘤技术获得了1株抗人IgE的单克隆抗体178,它具有IgE抗体特异性反应,其识别的抗原表位位于IgE与其受体结合的关键部位.经鉴定其具有体外阻断IgE与其受体结合的活性.结论 利用基因重组抗原制备了1株抗人IgE的单克隆抗体178,此单克隆抗体具有体外阻断IgE与其受体结合的生物活性.
目的 利用基因重組抗原免疫小鼠,製備抗人IgE單剋隆抗體,研究其特異性和功能.方法 用錶達的重組人IgE-Fc免疫BLAB/c小鼠,取其脾細胞與Sp2/0細胞融閤,以間接ELISA法篩選雜交瘤上清.用ELISA阻斷分析鑒定其生物學功能;通過丙氨痠突變掃描分析確定其抗原錶位.結果 利用雜交瘤技術穫得瞭1株抗人IgE的單剋隆抗體178,它具有IgE抗體特異性反應,其識彆的抗原錶位位于IgE與其受體結閤的關鍵部位.經鑒定其具有體外阻斷IgE與其受體結閤的活性.結論 利用基因重組抗原製備瞭1株抗人IgE的單剋隆抗體178,此單剋隆抗體具有體外阻斷IgE與其受體結閤的生物活性.
목적 이용기인중조항원면역소서,제비항인IgE단극륭항체,연구기특이성화공능.방법 용표체적중조인IgE-Fc면역BLAB/c소서,취기비세포여Sp2/0세포융합,이간접ELISA법사선잡교류상청.용ELISA조단분석감정기생물학공능;통과병안산돌변소묘분석학정기항원표위.결과 이용잡교류기술획득료1주항인IgE적단극륭항체178,타구유IgE항체특이성반응,기식별적항원표위위우IgE여기수체결합적관건부위.경감정기구유체외조단IgE여기수체결합적활성.결론 이용기인중조항원제비료1주항인IgE적단극륭항체178,차단극륭항체구유체외조단IgE여기수체결합적생물활성.
Objective To obtain monoclonal antibodies(McAb) against human IgE by recombinant IgE antigen.Methods BLAB/c mice were immunized with recombinant IgE.The spleen cells of immunized mice were used to prepare the McAb by hybridoma techniques.IgE-McAb was selected by indirect ELISA with culture supernatant of hybridoma cells.ELISA blocking assay were employed to identify the function of McAb.Alanine scanning mutagenesis were used to confirm the antigenic epitopes of McAb.Results A monoclonal antibody against IgE(178) was produced.The antigenic epitope of 178 was found to be spatially close to the receptor-binding site.Functional assay revealed that 178 could markedly inhibit IgE binding to receptors.Conclusion A hybridoma cell line secreting IgE-McAb stably was obtained by recombinant IgE antigen,and the IgE-McAb might block IgE binding to receptors via steric hindrance.