人促甲状腺激素单克隆抗体的制备及鉴定
Preparation and Identification of Monoclonal Antibody Against h-Thyroid-stimulating Hormone
乔元彪 1王权 2张文举 1陈沁1
作者信息
- 1. 上海大学,200444
- 2. 中国农业科学院上海兽医研究所,200241
- 折叠
摘要
目的:制备特异性强、灵敏度高的人促甲状腺激素(hTSH)单克隆抗体,为建立高灵敏性和高特异性的hTSH检测方法奠定基础.方法:人工合成人促甲状腺激素(hTSH)上的28个氨基酸,采用戊二醛交联法与牛血清白蛋白(BSA)构建完全抗原.用人工免疫原免疫BALB/c小鼠,利用杂交瘤技术建立分泌抗hTSH单克隆抗体的杂交瘤细胞,并对得到的单抗进行鉴定.结果:建立了一株杂交瘤细胞4E5,制备了腹水单抗,经鉴定,抗体重链属于IgG1类型,轻链属于κ型,McAb腹水ELISA效价达1:1.6×105,与促黄体生成激素(LH)、促卵泡激素(FSH)、人绒毛膜促性腺激素(HCG)无明显交叉反应,细胞株冻存复苏后抗体分泌稳定.结论:本实验建立的细胞株显示稳定性好,特异性高,McAb腹水效价高,为进一步提高hTSH检测的灵敏性和简便性提供依据.
Abstract
Objective To prepare high quality monoclonal antibody against human thyroid stimulating hormone(h-TSH) for high-sensitivity detection. Methods After synthetic hTSH β chain of 28 amino acids, glutaraldehyde cross-linked method was adopted to synthesize the complete antigen with bovine serum albumin. Through immunization of BALB/c mice with artificial immunogen, a hybri-doma cell line, which could produce monoclonal antibody(McAb) against hTSH was established. Results A hybridoma cell 4E5 was establised. Besides, prepared ascitic monoclonal antibody. It was proved that the heavy chain and light chain of hTSH McAb were IgGl and K type, respectively. The ELISA titer of ascites was 1:1.6×105. No significant cross-reactivity with LH, FSH, HGC. Hyhrido-mas cell line retained great stability after the freezing- thawing process. Conclusion The established cell lines showed high stability, high specificity and high titer of ascites, provide the basis for improving the hTSH detection sensitivity and simplicity.
关键词
人促甲状腺激素/单克隆抗体/杂交瘤细胞株Key words
hTSH/monoclonal antibodies (McAb)/hybridoma cell line引用本文复制引用
出版年
2013
NETL
NSTL
维普
万方数据