中文摘要：

目的：制备抗人脑钠肽单克隆抗体并对其进行初步鉴定。 方法：实验于2004-12／2006-02在南京医科大学第一附属医院心血管病研究所进行。利用碳化二亚胺法将合成的人脑钠肽与牛血清白蛋白偶联制备完全抗原，反复免疫Balb／c小鼠，应用杂交瘤技术将免疫小鼠的脾细胞与Balb／c小鼠骨髓瘤细胞（SP2／0）在PEG下融合，HAT选择性培养，间接酶联免疫吸附方法对细胞培养上清检测、筛选，选择筛选结果脑钠肽抗体阳性的细胞株连续进行3次亚克隆，建立稳定分泌脑钠肽单克隆抗体的杂交瘤细胞株。扩大培养阳性单克隆细胞株后，腹腔注射小鼠，制备腹水。对腹水亲和层析纯化，所得的单克隆抗体行初步鉴定。 结果：免疫小鼠血清呈脑钠肽抗体阳性，经筛选得到3株稳定分泌脑钠肽单克隆抗体的杂交瘤细胞株，纯化的单克隆抗体经鉴定为IgG型，蛋白质印迹分析证实单克隆抗体可特异识别脑钠肽，具备较高的特异性及敏感性。 结论：成功制备了3株抗脑钠肽特异性的单克隆抗体，可用于脑钠肽免疫检测方法的建立。

英文摘要：

AIM： To prepare monoclonal antibody （McAb） against human brain natriuretic peptide （BNP）, and perform primary identification. METHODS： The experiment was carried out from December 2004 to December 2006 at the Institute of Cardiovascular Disease, the First Affiliated Hospital of Nanjing Medical University. The synthesized BNP was conjugated to bovine serum albumin （BSA） using the coupling procedure in order to prepare complete antigen by carbediimide method, and then the conjugate was injected repeatedly into Balb/c mice as immunogen. Splenic cells of the immunized mice were fused in PEG with Balb/c mice myeloma cells （SP2/0）, and HAT selectively cultured. The hybridoma culture supemants were screened by indirect enzyme-linked immunoadsorbent assay （ELISA）. The cell strains positive for BNP antibody were selected for further use through three successive subclones to establish hybridoma cell strains, which excreted BNP McAb stably. These hybridoma cell strains were injected intraperitoneally into Balb/c mice for production of ascites. Each ascites was purified with affinity chromatography, and then the obtained McAb was evaluated by Western blot to test the biological activity. RESULTS： The serum of immunized mice was positive for BNP antibody. Three hybridoma cell strains were selected and excreted BNP McAb stably, and the purified McAb was determined as IgG and proved to identify BNP specifically in Western blot analysis, with high affinity, specificity and sensibitity. CONCLUSION： The McAbs against BNP with high activity and specificity have been established successfully, which can provide a potential value for establishing a new method to measure the serum concentrations of BNP.