中国农业科学院农业信息研究所 中国农学会
Agricultural Information Institute of CAAS China Association of Agricultural Science Societies
En
Ch
中国农业科学院农业信息研究所 中国农学会 主办

AI检索

订阅

返回顶部
当前位置: 首页 > 文章 > 抗重金属Cr~(3+)单克隆抗体的研制及其免疫学特性分析 西北农业学报 2016,25 (11) 1597-1602
Position: Home > Articles > Preparation of Anti-Chromium Ion Monoclonal Antibodies and Analysis of Immunological Properties Acta Agriculturae Boreali-occidentalis Sinica 2016,25 (11) 1597-1602

抗重金属Cr~(3+)单克隆抗体的研制及其免疫学特性分析

作  者：
王亚楠;丁菡;王晓斐;雷壮;牛琳琳;王自良
单  位：
河南科技学院动物科学学院;河南科技学院新科学院
关键词：
Cr3+;人工免疫原;单克隆抗体;免疫学特性
摘  要：
旨在研制敏感、特异、稳定的抗Cr~(3+)单克隆抗体(Cr~(3+)mAb)。用异硫氰酸苄基乙二胺四乙酸(ITCBE)鳌合Cr~(3+)形成Cr~(3+)-ITCBE半抗原,异硫氰酯法制备人工免疫原Cr~(3+)-ITCBE-BSA和检测抗原Cr~(3+)-ITCBE-OVA,紫外扫描法(UV)、凝胶电泳法(SDS-PAGE)和电感耦合等离子体原子发射光谱法(ICPAES)进行免疫原鉴定;用Cr~(3+)-ITCBE-BSA免疫Balb/C小鼠,细胞融合技术建立Cr~(3+)mAb杂交瘤细胞株,体内诱生腹水法制备Cr~(3+)mAb,并对其免疫学特性进行分析。结果表明,免疫原制备成功,Cr~(3+)-ITCBE-BSA中Cr~(3+)和BSA的质量浓度分别为140.8mg/L和5.81g/L,筛选出2A3C11、2A3D9、2A11G5 3株杂交瘤细胞,其中最好的2A11G5间接ELISA效价细胞培养上清为1∶(2.56×103)、腹水为1∶(5.12×105),同种型为IgG1/κ,亲和常数(Ka)为2.69×109 L/mol,对Cr~(3+)的半数抑制质量浓度(IC50)为16.6μg/L,与其他重金属离子无交叉反应。表明成功研制亲和力高、特异性强、稳定性好的Cr~(3+)mAb,为铬离子残留免疫学检测方法的建立奠定基础。
译  名：
Preparation of Anti-Chromium Ion Monoclonal Antibodies and Analysis of Immunological Properties
作  者：
WANG Yanan;DING Han;WANG Xiaofei;LEI Zhuang;NIU Linlin;WANG Ziliang;Animal Science College,Henan Institute of Science and Technology;School of Xinke,Henan Institute of Science and Technology;
关键词：
Chromium ion;;Artificial immunogen;;Monoclonal antibody;;Immunological properties
摘  要：
To develop sensitive,specific and stable monoclonal antibody against Cr~(3+)(Cr~(3+)mAb).Artificial hapten Cr~(3+)-ITCBE was synthesized to chelate chromium ion(Cr~(3+))using ITCBE.The method of isothiocyanate(ITC)was used to conjugate Cr~(3+)-ITCBE to bovine serum albumin(BSA)and artificial immunogen Cr~(3+)-ITCBE-BSA was obtained,the detecting antigen Cr~(3+)-ITCBE-OVA was obtained in the same way.Ultraviolet(UV),sodium dodecyl sulfate polyacrylamide gel electropheresis(SDS-PAGE)and inductively coupled plasma atomic emission spectroscopy(ICP-AES)were used to identify Cr~(3+)-ITCBE-BSA.Balb/C mice were immunized with Cr~(3+)-ITCBE-BSA and hybridoma lines that secrete monoclonal antibody against Cr~(3+)(Cr~(3+)mAb)were established through cell fusion,Cr~(3+)mAb were induced from in vivo method and and their immunological properties of Cr~(3+)mAb were identified.The results showed that Cr~(3+)-ITCBE-BSA was synthesized successfully and its mass concentration of Cr~(3+)and BSA was 140.8 mg/L and 5.81g/L respectively.Three hybridoma lines of 2A3C11,2A3D9,2A11G5 were screened and the best one was 2A11G5 with indirect ELISA titers of 1∶(2.56×103)in supernatant and 1∶(5.12×105)in ascites.The isotype of the mAb was IgG1/κand its affinity constant(Ka)was 2.69×109 L/mol.The mAb showed good sensitivity with an IC50 of 16.6μg/L to Cr~(3+)and little or no cross-reactivity with other compounds.This study had successfully developed the Cr~(3+)mAb,which had high affinity,specificity and stability,and provided a potency of establishing immunoassay methods for determining Chromium ion residues in environment and food.

相似文章

导出引用
计量
文章访问数: 8
HTML全文浏览量: 0
PDF下载量: 0

所属期刊

推荐期刊

友情链接Friendly link
农业信息研究所Agricultural information institute 农业智能知识服务平台Agricultural intelligent knowledge service platform 农业学术服务平台Agricultural Knowledge Service System 中国知网CNKI 维普网VIP 万方WANFANG DATA
联系我们Contact us

电话Tell：010-82109652-808

邮箱E-mail：agrijournal@caas.cn

服务Service
关于我们About us 意见建议Opinions and suggest
版权说明Copyright
单位：中国农业科学院农业信息研究所 中国农学会 © 2017中国农业科学院农业信息研究所 ICP备案号：京ICP备09089781号-46