CAJ | 학술논문

YycFG双组分信号转导系统调控葡萄球菌细胞壁合成、代谢及生物膜形成，PAS是组氨酸激酶YycG的胞外信号感应区域。本研究针对胞外PAS区域制备抗YycG‐PAS单克隆抗体（YycG‐PAS MAb ）。免疫鉴定结果表明，YycG‐PAS MAb可与重组YycG‐PAS蛋白和表皮葡萄球菌菌体 YycG蛋白结合，属于IgG2a亚型，效价为log 215。YycG‐PAS MAb（80μg／ml）与表皮葡萄球菌 RP62A菌株共培养24 h ，对细菌生物膜形成的抑制率达46．5％，与 mIgG （80μg／mL ）组相比有显著差异（ P＜0．05）。YycG‐PAS MAb （80μg／ml）与低浓度万古霉素（1μg／ml）联用可增强对24 h细菌生物膜的抑制作用，抑制率由57．6％提高至93．0％（P＜0．01）。YycG‐PAS MAb对浮游状态表皮葡萄球菌的生长及存活无显著影响。本研究为YycG‐PAS MAb在抗生物膜感染中的潜在应用价值奠定了一定基础。
YycFG쌍조분신호전도계통조공포도구균세포벽합성、대사급생물막형성，PAS시조안산격매YycG적포외신호감응구역。본연구침대포외PAS구역제비항YycG‐PAS단극륭항체（YycG‐PAS MAb ）。면역감정결과표명，YycG‐PAS MAb가여중조YycG‐PAS단백화표피포도구균균체 YycG단백결합，속우IgG2a아형，효개위log 215。YycG‐PAS MAb（80μg／ml）여표피포도구균 RP62A균주공배양24 h ，대세균생물막형성적억제솔체46．5％，여 mIgG （80μg／mL ）조상비유현저차이（ P＜0．05）。YycG‐PAS MAb （80μg／ml）여저농도만고매소（1μg／ml）련용가증강대24 h세균생물막적억제작용，억제솔유57．6％제고지93．0％（P＜0．01）。YycG‐PAS MAb대부유상태표피포도구균적생장급존활무현저영향。본연구위YycG‐PAS MAb재항생물막감염중적잠재응용개치전정료일정기출。
Bacterial YycFG two‐component signal transduction system plays a major role in controlling cell wall synthesis , metabolism and biofilm formation of Staphylococcus . The extracellular PAS domain of YycG histidine kinase can sense various environmental signals .Therefore ,a monoclonal antibody targeting PAS domain of Staphylococcus epidermidis (S . epidermidis) was produced (YycG‐PAS MAb) .Enzyme‐linked immunosorbent assay (ELISA ) indicated that YycG‐PAS MAb belonged to IgG2a subclass and antibody titer was log 215 .Western blotting suggested that YycG‐PAS MAb had the ability to bind both recombinant YycG‐PAS protein and protein extracted from cells .Further investigation exhibited inhibitory effect of YycG‐PAS MAb (80 μg/ml) against biofilm formation after co‐incubation with S . epidermidis and the inhibitory rate was 46 .5% . The anti‐biofilm activity of YycG‐PAS MAb was improved when combined with low‐concentration of vancomycin (1 μg/ml) .The inhibitory rate was increased from 57 .6% <br> to 93 .0% ( P< 0 .01 ) . No significant effects of YycG‐PAS MAb on viability and growth of planktonic bacterial cells were observed .This study indicated that YycG‐PAS MAb is a promising candidate antibody for biofilm‐related infections .