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人白血病抑制因子(hLIF)单克隆抗体的制备与鉴定
作者单位:1.昆明医科大学 分子临床医学研究院,2.云南大学 单克隆抗体技术中心,3.昆明医科大学 科研实验中心
摘    要: 为了获得抗人白血病抑制因子(hLIF)单克隆抗体,以重组hLIF蛋白为抗原,免疫Balb/c小鼠,采用传统的小细胞融合、无限稀释法制备了10株杂交瘤的腹水hLIF抗体,并对腹水进行了纯化.腹水粗品抗体经rProteinA一步亲和层析法纯化和鉴定后,抗体分子量都符合抗体的特性,抗体的纯度都达到了96.6%以上,抗体亲和常数在1.89×10-9~1.51×10-12mol/L,其中1B8a、5C1b、5G4a细胞株制备的抗体属于高亲和力抗体,可以对其的应用进行进一步的研究和探讨.

关 键 词:人白血病抑制因子(hLIF)  亲和常数  杂交瘤  单克隆抗体

Preparation and detection of anti-hLIF monoclonal antibodies
Institution:1.Institute of Molecular and Clinical Medicine,Kunming Medical University,2.Monoclonal Antibody Research Center,Yunnan University,3.Scientific Research Experiment Center,Kunming Medical University
Abstract:To generate monoclonal antibody against human leukemia inhibitory factor,Balb/c mice were immunized with hLIF protein and the 10 of hybridomas were obtained by using tranditional cell fusion and limited dilution techniques.The ascites of 10 hybridomas were purified via rProteinAone-step affinity chromatography,The antibodies were determinated,in which the antibodies-molecular weight were in line with antibody puoperties,its purity was 96.6% and above,antibody affinity constant was in 1.89×10-9~1.51×10-12mol/L,especially 1B8a,5C1b,5G4a cell preparation-antibody showed to the high affinity antibodies,so we can apply for further research and investigation.
Keywords:human leukemia inhibitory factor  affinity constant  hybridoma  monoclonal antibody
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