针对人 CYP11B1 和 CYP11B2 的单克隆抗体的开发。

Development of monoclonal antibodies against human CYP11B1 and CYP11B2.

Endocrine Section, G.V. (Sonny) Montgomery VA Medical Center, USA; Endocrinology, University of Mississippi Medical Center, USA.

Endocrinology, University of Mississippi Medical Center, USA.

Mol Cell Endocrinol. 2014 Mar 5;383(1-2):111-7. doi: 10.1016/j.mce.2013.11.022. Epub 2013 Dec 8.

The final enzymes in the biosynthesis of aldosterone and cortisol are by the cytochrome P450 CYP11B2 and CYP11B1, respectively. The enzymes are 93% homologous at the amino acid level and specific antibodies have been difficult to generate. 2. Mice and rats were immunized with multiple peptides conjugated to various immunogenic proteins and monoclonal antibodies were generated. The only peptide sequences that generated specific antibodies were amino acids 41-52 for the CYP11B2 and amino acids 80-90 for the CYP11B1 enzyme. 3. The mouse monoclonal CYP11B2-41 was specific and sensitive for use in western blots and produced specific staining of the zona glomerulosa of normal adrenal glands. The rat monoclonal CYP11B1-80 also detected a single band by western blot and detected only the zona fasciculata. Triple immunofluorescence of the adrenal demonstrated that the CYP11B1 and the CYP11B2 did not co-localize, while as expected the CYP11B1 co-localized with the 17α-hydroxylase.

醛固酮和皮质醇生物合成的最后两种酶分别为细胞色素 P450 CYP11B2 和 CYP11B1。这两种酶在氨基酸水平上有 93%的同源性，因此难以产生特异性抗体。
研究人员用多种与免疫原性蛋白偶联的肽段对小鼠和大鼠进行免疫，生成了单克隆抗体。唯一能产生特异性抗体的肽序列是 CYP11B2 的 41-52 位氨基酸和 CYP11B1 酶的 80-90 位氨基酸。
小鼠单克隆 CYP11B2-41 特异性和灵敏度高，可用于 Western blot，并对正常肾上腺的肾小球带产生特异性染色。大鼠单克隆 CYP11B1-80 也通过 Western blot 检测到一条带，且仅检测到束状带。肾上腺的三重免疫荧光显示，CYP11B1 和 CYP11B2 没有共定位，而预期的是，CYP11B1 与 17α-羟化酶共定位。

Development of monoclonal antibodies against human CYP11B1 and CYP11B2.