抗尿苷二磷酸葡萄糖焦磷酸化酶2单克隆抗体的制备与鉴定.docVIP

抗尿苷二磷酸葡萄糖焦磷酸化酶2单克隆抗体的制备与鉴定 目录 TOC \o 1-9 \h \z \u 目录 1 正文 1 文1：抗尿苷二磷酸葡萄糖焦磷酸化酶2单克隆抗体的制备与鉴定 1 文2：抗人绒毛膜促性腺激素单克隆抗体的制备与鉴定 7 1 材料和方法 7 2 结果 9 3 讨论 10 参考文摘引言： 11 原创性声明（模板） 12 文章致谢（模板） 13 正文 抗尿苷二磷酸葡萄糖焦磷酸化酶2单克隆抗体的制备与鉴定 文1：抗尿苷二磷酸葡萄糖焦磷酸化酶2单克隆抗体的制备与鉴定 Preparation and Identification of Monoclonal Antibody Agait Abstract The study was aimed to generate monoclonal antibodies (mAbs) agait homo sapie UDPglucose pyrophosphorylase 2 (UGP2). Normal human liver tissues homogenized, and cytosolic protei isolated by centrifugation were used to immunize BALB/c mice to generate mAbs by hybridoma technique. The mAbs were identified by ELISA, Western blot, and immunohistochemistry assay. The antibody specificity was confirmed by UniZAP expression library screening. The results indicated that one hybridoma BAD062 secreting specific mAb agait UGP2 was established. The Ig subclass of this mAb was IgG2b (κ), and it could be used in ELISA, Western blot, immunohistochemistry assay. The antigen recognized by BAD062 mAb was localized in the hepatocyte cytoplasm, with molecular weight of 56 kD in the cytosolic protei of human liver tissue. The BAD062 mAb was further confirmed by immunoscreening of UniZAP XR liver cDNA expression library. It is concluded that a hybridoma cell line stably secretes specific mAb agait UGP2. This mAb reacted with UGP2 in ELISA, Western blot, immunohistochemistry assay, and would be very useful for the UGP2 studies. Key words UGP2; mAb; cDNA expression library 尿苷二磷酸葡萄糖焦磷酸化酶2（UDPglucose pyrophosphorylase2, UGP2）是糖原生物合成过程中的一种酶，由508个氨基酸组成，相对分子质量（Mr）约为56 000，催化UTP与葡萄糖1P的反应, 生成UDP葡萄糖。UDP葡萄糖是主要活化糖的形式，作为葡萄糖基供体参与蔗糖、纤维素、半纤维素、果胶质以及糖脂、糖蛋白的合成代谢［1］。摄入人体内的葡萄糖可在葡萄糖激酶、葡糖磷酸变位酶和尿苷二磷酸葡糖焦磷酸化酶（UGP2）的催化下形成尿苷二磷酸葡萄糖（UDPG）。此外，UGP2还与腺嘌呤二磷酸葡萄糖焦磷酸化酶（AGPase）相偶联，形成ADPG，参与淀粉的合成［1,2］。在糖原合成和分解过程中该酶的缺陷即导致临床表现的糖原累积病。UGP2在骨骼肌中高表达，其次在肝脏中也有较高的表达量。我们应用成人肝组织胞浆总蛋白免疫BALB/c小鼠制备单克隆抗体，其中1株为鼠抗人UGP2单克隆抗体，并对其特异性进行了鉴定。这为进一步研究UGP2在糖蛋白代谢中的作用奠定了基础。 中国 实验血液学杂志 J Exp Hematol 2007; 15(3) 抗尿苷