人促甲状腺激素单克隆抗体的制备及鉴定

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K型，McAb腹水ELISA效价达1：1．6×10^5，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。

Preparation and Identification of Monoclonal Antibody Against h-Thyroid-stimulating Hormone

Objective To prepare high quality monoclonal antibody against human thyroid stimulating hormone（h-TSH） for highsensitivity detection. Methods After synthetic hTSH 13 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 hybridoma 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 IgG1 and K type, respectively. The ELISA titer of ascites was 1：1.6×10^5. No significant cross-reactivity with LH, FSH, HCG. Hybridomas cell line retained great stability after the freezingthawing 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.