CAJ | 학술논문

目的本研究拟探讨1株黄病毒属特异的单克隆抗体2A10作为检测抗体在同时检测5种黄病毒属病毒的蛋白芯片中的应用价值.方法将病毒的特异性单抗作为捕获抗体进行点样,制备用于检测5种黄病毒属病毒的抗体芯片,然后分别加入拟检测的病毒灭活抗原,捕获病毒颗粒后,加入荧光染料CY3标记的2A10检测抗体进行孵育,通过扫描并分析芯片上不同抗体的荧光强度,确定2A10检测抗体的效价和可检测的病毒滴度,并对2A10的检测效果进行评价.结果蛋白芯片检测结果表明,2A10可作为检测抗体用于对5种黄病毒属病毒的检测,CY3标记的2A10效价为1∶160,以脾传脑炎(TBE)病毒为例,可检测的病毒滴度为1250 PFU.结论单克隆抗体2A10作为一株黄病毒属特异的单抗,可以用于蛋白芯片技术检测黄病毒属病毒.
목적 본연구의탐토1주황병독속특이적단극륭항체2A10작위검측항체재동시검측5충황병독속병독적단백심편중적응용개치.방법 장병독적특이성단항작위포획항체진행점양,제비용우검측5충황병독속병독적항체심편,연후분별가입의검측적병독멸활항원,포획병독과립후,가입형광염료CY3표기적2A10검측항체진행부육,통과소묘병분석심편상불동항체적형광강도,학정2A10검측항체적효개화가검측적병독적도,병대2A10적검측효과진행평개.결과 단백심편검측결과표명,2A10가작위검측항체용우대5충황병독속병독적검측,CY3표기적2A10효개위1∶160,이비전뇌염(TBE)병독위례,가검측적병독적도위1250 PFU.결론 단극륭항체2A10작위일주황병독속특이적단항,가이용우단백심편기술검측황병독속병독.
Objective To evaluate the Flavivirus specific monoclonal antibody(McAb) 2A10 as detective antibody for simultaneously identify tick borne encephalitis virus( TBEV), Japanese encephalitis virus( JEV), dengue ( DEN )-2, DEN-4 and yellow fever virus ( YFV ) by antibody microarray technique.Methods The antibody microarray was developed by spotting TBEV, JEV, DEN-2, DEN-4 and YFV specific McAb on chip as capture antibodies. After incubating with cultured viral supernatants of the above viruses, CY3 labeled detective antibody 2A10 was added to the chips. After reaction, the antibody microarray was scanned and the results were analyzed. By comparing the signal intensities of different spots on chips,the detecting titre and sensitivity of 2A10 for Flavivirus were determined, and the value of 2A10 in detection of Flavivirus was evaluated. Results The hybridization results demonstrated that the titre of 2A10 for Flavi2A10 was specific for Flavivirus and could be used as universal detective antibody for Flavivirus on antibody microarray.