Chen Yanliang, Zhang Chaoyue
Department of Orthopedics, Third Xiangya Hospital of Central South University , Changsha, China .
Monoclon Antib Immunodiagn Immunother. 2016 Jun;35(3):163-6. doi: 10.1089/mab.2015.0069. Epub 2016 May 26.
Spermidine/spermine N1-acetyltransferase (SSAT) is a catabolic regulator of polyamines, ubiquitous molecules essential for cell proliferation and differentiation. Anti-SSAT antibodies (monoclonal antibodies [mAbs]) of high titer were prepared by immunizing BALB/c mice with multifocal intradermal injections and by fusing high-titer antibody-producing spleen cells with myeloma cells of SP2/0 origin. Four mAbs were selected for further characterization as classes and subclasses. Antibodies were produced by these three clones with high affinities ranging from 10(9) to 10(11) M(-1). These clones were found to be of the immunoglobulin IgG1 subclass with kappa light chain. They could recognize SSAT as determined by Western blot and immunohistochemistry. The specificity of one clone, 4H6, was studied by using the small interfering RNA (siRNA) on SSAT. 4H6 was also compared with the commercial antibody. The produced mAbs will be a useful tool for further investigation of SSAT functions in organisms.
亚精胺/精胺N1-乙酰基转移酶（SSAT）是多胺的分解代谢调节因子，多胺是细胞增殖和分化所必需的普遍存在的分子。通过多灶性皮内注射免疫BALB/c小鼠，并将产生高滴度抗体的脾细胞与SP2/0来源的骨髓瘤细胞融合，制备了高滴度的抗SSAT抗体（单克隆抗体[mAbs]）。选择了四种单克隆抗体进行进一步的类别和亚类鉴定。这三个克隆产生的抗体具有10⁹至10¹¹ M⁻¹的高亲和力。这些克隆被发现属于具有κ轻链的免疫球蛋白IgG1亚类。通过蛋白质印迹和免疫组织化学测定，它们可以识别SSAT。使用针对SSAT的小干扰RNA（siRNA）研究了一个克隆4H6的特异性。还将4H6与商业抗体进行了比较。所产生的单克隆抗体将成为进一步研究SSAT在生物体中功能的有用工具。
