乙肝病毒表面抗原G145R变异单克隆抗体的制备及其初步鉴定

来源 :细胞与分子免疫学杂志 | 被引量 : 0次 | 上传用户:mengyidaocaoren
下载到本地 , 更方便阅读
声明 : 本文档内容版权归属内容提供方 , 如果您对本文有版权争议 , 可与客服联系进行内容授权或下架
论文部分内容阅读
目的:研制抗乙肝病毒G145R变异表面抗原的单克隆抗体(mAb)。方法:将重组乙肝病毒G145R变异表面抗原以Al(OH)3佐剂化,常规免疫BALB/c小鼠,采用细胞融合技术制备抗乙肝病毒G145R变异表面抗原的mAb。采用ELISA对制备物的效价与特异性进行鉴定,将其用于部分临床标本的检测,并与德国抗HBs mAb GL2.001进行比较。结果:获得1株稳定分泌抗G145R HBs mAb的杂交瘤细胞,命名为4-E12。该细胞株染色体数目为90条,所分泌mAb属IgG1亚类。其培养上清与腹水抗G145R HBs ELISA效价为(0.5~1)×103及(1~10)×106。经持续3月培养,低血清适应以及反复冻存与复苏后,其细胞增殖能力与上清抗G145R HBs效价与克隆化结束时大致相似。将其与德国抗G145R HBs mAb GL2.001相比,二者对重组乙肝表面抗原G145R变异S蛋白的反应性大致相当,灵敏度前者稍低于后者(分别为2.0μg/L与1.0μg/L)。对野生株表面抗原的检测前者在实验浓度范围内未显示有意义的反应信号,后者在与高浓度(1mg/L及以上)抗原反应时S/N值达到或高于3.9。将二者用于对一组特定患者的临床检测,其阳性率前者(17/200,8.5%)较后者(11.5%)略低,其差别经统计学比较无统计学意义(P>0.05)。结论:成功地制备了1株抗乙肝表面抗原G145R变异体的mAb,为乙肝G145R变异的基础、临床与流行病学研究提供了进一步的物质基础。 Objective: To develop a monoclonal antibody (mAb) against hepatitis B virus G145R variant surface antigen. Methods: The recombinant hepatitis B virus G145R mutant surface antigen was adjuvated with Al (OH) 3, and BALB / c mice were immunized routinely. The mAbs against hepatitis B virus G145R variant surface antigen were prepared by cell fusion technique. The titer and specificity of the preparations were identified by ELISA, which were used for the detection of some clinical specimens and compared with the German anti-HBs mAb GL2.001. Results: One hybridoma cell stably secreting anti-G145R HBs mAb was obtained and named as 4-E12. The number of chromosomes in this cell line is 90, and the secreted mAb belongs to IgG1 subclass. The titers of anti-G145R HBs in culture supernatants and ascites were (0.5 ~ 1) × 103 and (1 ~ 10) × 106. After continued cultivation in March, low serum adaptation and repeated cryopreservation and resuscitation, the cell proliferation ability and the supernatant anti-G145R HBs titers were similar to those at the end of cloning. Compared with the German anti-G145R HBs mAb GL2.001, the reactivity against the recombinant hepatitis B surface antigen G145R mutant S protein was approximately equivalent, with the former slightly lower than the latter (2.0 μg / L and 1.0 μg / L respectively ). Detection of wild-type surface antigen The former did not show significant response signals within the experimental concentration range, with S / N values ​​at or above 3.9 when reacted with high concentrations of antigen (1 mg / L and above). The two were used for clinical testing of a specific group of patients, the positive rate (17/200%, 8.5%) was slightly lower than that of the latter (11.5%). The difference was statistically insignificant (P> 0.05 ). CONCLUSION: A mAb of G145R variant of hepatitis B surface antigen was successfully prepared, which provided the basis for further study of G145R mutation in hepatitis B virus. The clinical and epidemiological studies provided a further material basis.
其他文献
为加快“十一五”期间全省林业人才队伍建设,从根本上改善林业人才队伍的现状,必须着力抓好几项工作。切实加强对林业人才工作的领导各级林业部门要把人才工作作为林业建设的
期刊
由于经济的快速增长,1978年以来的25年中,珠江三角洲地区的土地覆盖变化,特别是城市扩展速度非常之快.通过把遥感和地理信息系统相结合,研究了土地覆盖变化及城市增长状况,并
只知楚越为天涯, 此身如线自萦绕, 误抛山林入朝市, 欲从稚川隐罗浮, 首参虞舜款韶石, 仙山一见五色羽,
浙江省建德市下涯镇党委书记胡解放受贿“东窗事发”,市检察院这边讯问被告人,那边却有人向犯罪嫌疑人通风报信。而作奸犯科者竟是该院个别检察官。这种事件在建德市自然已非首
毓麟珠方出自<景岳全书*妇人规>,由人参、白术、茯苓、芍药、川芎、甘草、当归、熟地黄、菟丝子、杜仲、鹿角霜、川椒组成.笔者运用该方治疗排卵障碍[1] 时,偶然发现该方对女
介绍核磁共振的基本原理和故障维修。
利用MODIS数据的分裂窗算法,获取了浙江省各地2008年一整年的地表温度数据,综合利用遥感、地理信息系统(GIS)技术,研究了地表温度的时空分布特征、地表温度与不同土地覆盖类型
本文用基于最小位能原理的有限元方法研究了直升机悬停时旋翼浆叶挥舞—摆振耦合颤振。利用准定常二维片条理论求出气动载荷。由非线性运动方程求出定常状态浆叶变形,假定浆
春季保护地黄瓜栽培使用穴盘育苗技术,可以缩短缓苗期,提高成活率,避免低温等不利环境条件对苗期生长的影响。现将早春黄瓜穴盘育苗关键技术总结如下。1基质的选择与苗床建造
在加快改革开放和加速经济发展的新形势下,有一些党组织在实际工作中往往对干部中的问题睁一只眼,闭一只眼,只要上级不追查,能避开的则避开,致使有些问题长期得不到解决。从
期刊