CAJ | 학술논문

目的制备针对热休克蛋白27/热休克蛋白B1(HSP27/HSPB1)的特异性单克隆抗体并对其进行鉴定.方法用HSP27/HSPB1作为免疫抗原免疫BalB/c小鼠,通过常规杂交瘤技术将免疫后的小鼠脾细胞与SP2/0细胞经PEG融合,筛选阳性杂交瘤细胞株,进行克隆化培养后接种于小鼠腹腔制备HSP27/HSPB1单克隆抗体,并利用间接酶联免疫吸附试验、免疫印迹、免疫组织化学、免疫荧光技术对所制备的抗体进行鉴定.结果获得了一株可稳定分泌HSP27/HSPB1蛋白单克隆抗体的杂交瘤细胞株,命名为4C9C7,其亚类是IgG1,分泌的单克隆抗体能特异性结合组织和细胞中的HSP27/HSPB1蛋白.结论本实验成功制备了可稳定分泌HSP27/HSPB1单克隆抗体的杂交瘤细胞及HSP27/HSPB1特异性的单克隆抗体,可应用于各种相关的科学研究.
목적제비침대열휴극단백27/열휴극단백B1(HSP27/HSPB1)적특이성단극륭항체병대기진행감정.방법용HSP27/HSPB1작위면역항원면역BalB/c소서,통과상규잡교류기술장면역후적소서비세포여SP2/0세포경PEG융합,사선양성잡교류세포주,진행극륭화배양후접충우소서복강제비HSP27/HSPB1단극륭항체,병이용간접매련면역흡부시험、면역인적、면역조직화학、면역형광기술대소제비적항체진행감정.결과획득료일주가은정분비HSP27/HSPB1단백단극륭항체적잡교류세포주,명명위4C9C7,기아류시IgG1,분비적단극륭항체능특이성결합조직화세포중적HSP27/HSPB1단백.결론본실험성공제비료가은정분비HSP27/HSPB1단극륭항체적잡교류세포급HSP27/HSPB1특이성적단극륭항체,가응용우각충상관적과학연구.
Objective To prepare and indentify specific monoclonal antibody (McAb) against heat shock protein 27/heat shock protein B1 (HSP27/HSPB1). Methods BalB/c mice were immunized with purified HSP27/HSPB1 protein expressed by prokaryotic expression system. Then the spleen cells from immunized mice were fused with SP2/0 myeloma cells by conventional hybridoma technique. The positive hybridoma cells were chosen to clone and culture, and then were injected into the abdominal cavities of BalB/c mice. The prepared antibody was identified by indirect enzyme linked immunosorbent assay (ELISA), western blotting, immunohistochemistry and immunofluorescence technique. Results A hybridoma cell line that was able to stably secrete HSP27/HSPB1 monoclonal antibody was obtained and named 4C9C7. Its subtype was IgG1. Its secreted antibodies could spe-cifically bind to HSP27/HSPB1 protein from different types of tissues and cells. Conclusion A hybridoma cell line that was capa-ble to stably secret HSP27/HSPB1 monoclonal antibody was successfully prepared in our study and it may be applied to various related scientific researches.