Development and characterization of a mouse monoclonal antibody against antimicrobial peptide tachyplesin I.
Development and characterization of a mouse monoclonal antibody against antimicrobial peptide tachyplesin I.
复制标题
抗抗菌肽鲎素 I 的小鼠单克隆抗体的开发和表征。
作者：
Xin Feng;W. Han;Zhanyun Song;Honglei Zhao;Yu Gao;Yuwen Diao;Shanshan Liu;L. Lei
为研究其体内、外阳离子抗菌肽(AMPs)的活性机制，并从表达产物中分离纯化TP I，研制了抗TP I的单抗。将合成的TP I与载体蛋白BSA化学偶联，然后注射到BALB/c小鼠体内。用TP I间接酶联免疫吸附试验(ELISA法)筛选阳性杂交瘤细胞，有限稀释法进行3次亚克隆。获得了5株能有效检测天然TP I的单抗(命名为2D8、3B8、5H2、6B12和8F5)。单抗2D8、3B8、5H2和8F5属于IgG1，单抗6B12属于IgG2a。特异性试验表明，单抗8F5对天然TP I、重组TP I、合成TP I和TP II的特异性与天然TP I、重组TP I、合成TP I和TP II几乎相同，但与对照多肽无交叉反应。这些结果表明，合成的AMP偶联物可以诱导针对天然AMP的抗体，并可用于抗菌肽的检测。
Monoclonal antibodies against tachyplesin I (TP I) were developed to study its mechanisms of activity, a kind of cationic antimicrobial peptides (AMPs), in vivo or in vitro, and to purify TP I from expression products. The synthesized TP I was chemically conjugated with the carrier protein BSA and then injected into BALB/c mice. Positive hybridomas were screened by indirect enzyme-linked immunosorbent assay (ELISA) using TP I and subcloned three times with limiting dilution. Five MAbs effective in detecting the native TP I (named 2D8, 3B8, 5H2, 6B12, and 8F5) were obtained. Isotyping of all obtained MAbs indicated that MAbs 2D8, 3B8, 5H2, and 8F5 belong to IgG1, and MAb 6B12 belongs to IgG2a. Specificity assay showed that MAb 8F5 had almost the same level of specificity to natural TP I, recombinant TP I, and synthesized TP I and TP II, but did not cross-react with control peptides. These results suggest that the synthetic AMP conjugates can elicit antibodies against native AMPs and can be used to detect antimicrobial peptides.
影响因子：
3.3
作者：
Nguyen, Khoi Tan;King, John Thomas;Chen, Zhan
通讯作者：
Chen, Zhan
影响因子：
11.2
作者：
Chen, JG;Xu, XN;Zhang, LR
通讯作者：
Zhang, LR
影响因子：
11.2
作者：
Yixin Chen;Xue-Ming Xu;Shuigen Hong;Jinguo Chen;Ningfei Liu;Charles B. Underhill;Karen Creswell;Lurong Zhang
通讯作者：
Yixin Chen;Xue-Ming Xu;Shuigen Hong;Jinguo Chen;Ningfei Liu;Charles B. Underhill;Karen Creswell;Lurong Zhang