| 关雪 刘福剀 杨昕蕾 陈剑锋 杨永良 梁重阳.抗人beta-HCG单克隆抗体的制备及化学发光
检测方法的建立[J].,2014,14(1):68-73 |
| 抗人beta-HCG单克隆抗体的制备及化学发光
检测方法的建立 |
| Preparation of Monoclonal Antibodies Against Human beta-HCGand Establishment of a Sandwich CLIA System |
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| DOI： |
| 中文关键词: 人绒毛膜促性腺激素（beta-HCG） 单克隆抗体 夹心CLIA |
| 英文关键词: Human chorionic gonadotrophin (beta-HCG) Monoclonal antibody Sandwich CLIA |
| 基金项目:国家自然科学基金青年基金项目(81202446) |
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| 中文摘要: |
| 目的：制备人绒毛膜促性腺激素（beta-HCG）单克隆抗体，建立人beta-HCG双抗体夹心CLIA 检测方法。方法：用人beta-HCG 抗原
免疫小鼠，通过细胞融合、筛选后得到杂交瘤细胞株，然后将细胞株扩大培养并纯化上清液获得抗体，测定抗体亲和力、特异性及
表位，最后建立双抗体夹心CLIA方法。结果：获得4 株抗人茁-HCG的杂交瘤细胞株(beta-1-1、beta-2-1、beta-3-1、beta-4-1)。用beta-1-1 和
beta-2-1 建立的双抗体夹心法的检测范围为0.5 mIU/mL-800 mIU/mL，灵敏度0.23 mIU/mL，检测结果的相对偏差均在± 10 %内，回
收率在90 %以上。结论：本研究最终成功制备了抗人beta-HCG mAb，建立了定量检测人beta-HCG 的双抗体夹心CLIA 方法，为
beta-HCG 检测及疾病的诊断奠定基础。 |
| 英文摘要: |
| Objective: To prepare human chorionic gonadotrophin (beta-HCG) monoclonal antibody, and establish double antibody
sandwich CLIA detecting system for human beta-HCG. Methods: Mice were immunized with human beta-HCG antigen, and then hybridoma
cell lines were obtained by cell fusion and screened for the clones which expressed the antibody. The positive cell clones were expanded
further for a large volume of supernatant, from which the antibodies were purified and tested for affinity, specificity and epitope. Finally
the obtained antibodies were used to establish the double antibody sandwich CLIA system. Results: 4 monoclones of anti-human beta-HCG
(beta-1-1, beta-2-1, beta-3-1, beta-4-1) antibody were obtained. beta-1-1 and beta-2-1 were used to establish the double antibody sandwich system with
detection range of 0.5 mIU/mL-800 mIU/mL, sensitivity of 0.23mIU/mL, relative detection deviation of ± 10%and recovery rate of over
90% .Conclusion:The double antibody sandwich CLIA system for quantitative detection of human beta-HCG, using the prepared
anti-human beta-HCG mAbs, was successfully established, which lay the foundation for the diagnostic beta-HCG detection in the diseases.
Human chorionic gonadotrophin (beta-HCG); Monoclonal antibody; Sandwich CLIA |
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