人源性TPO单克隆抗体杂交瘤细胞株的建立

ESTABLISHMENT OF MONOCLONAL ANTIBODIES AGAINST HUMAN THROMBOPOIETIN

用人源性ＴＰＯ（ｈｕＴＰＯ）皮下免疫ＢＡＬＢ／ｃ小鼠，取脾细胞与ＳＰ２／０实体瘤细胞融合，建立了２株稳定分泌抗ＴＰＯ单克隆抗体的杂交瘤细胞株，分别命名为１９Ｄ１２和４９Ａ５，并对其进行了初步鉴定，采用简易正辛酸法从腹水中纯化单克隆抗体。结果：细胞融合率为８２％，杂交瘤细胞染色体数目正常，分泌的抗体为ＩｇＧ，腹水抗体效价为０．６～１．２×１０－５，与ＥＰＯ、ＧＭ－ＣＳＦ无交叉反应，连续培养生物性状稳定。抗ＴＰＯ单克隆抗体杂交瘤细胞株的建立。

To prepare monoclonal antibody(McAb) against human thrombopoietin(huTPO),BALB/c mice were immunized with purified huTPO. Hybridomas were made using polythylene glycol(PEG)as the fusing agent.Two hybridoma cell lines,19D12 and 49A5,were obtained.The McAb was purified from ascites by a simple procedure of caprylic acid precipitation.The rate of fusion was 82 percent.The chromosome counts of hybridoma cell was normal and the Ig class of McAb was IgG.The titer of the TPO McAbs in the ascites was 0.6～1.2×10-5 in ELISA.The McAbs against human TPO may be useful tool for studing biologic properties of TPO and clinical laboratory detection.