CAJ | 학술논문

制备高特异性的抗鲤春血症病毒单克隆抗体，并对其免疫学特性进行鉴定。采用差速离心法提纯的鲤春血症病毒作为免疫原，免疫Balb/c小鼠，4次免疫后，利用杂交瘤细胞技术进行细胞融合，经过多次亚克隆筛选出高敏感的特异性抗鲤春血症病毒单克隆抗体杂交瘤细胞株，体内诱生腹水制备抗鲤春血症病毒单克隆抗体并对其免疫学特性进行鉴定。融合后筛出两株杂交瘤细胞株9C11和1C12，亚型均为IgG2b型，κ链；间接ELISA测定腹水效价为104，与其他病毒株和细胞株不发生反应；western-blot结果显示单克隆抗体对应的抗原位点在相对分子质量（Mr）230000的条带处；免疫氧化酶染色结果显示单克隆抗体能够特异地识别感染细胞内的鲤春血症病毒。本实验制备的抗鲤春血症病毒单克隆抗体特异性好，为鲤春血症病毒免疫学快速检测方法的建立奠定了基础。
제비고특이성적항리춘혈증병독단극륭항체，병대기면역학특성진행감정。채용차속리심법제순적리춘혈증병독작위면역원，면역Balb/c소서，4차면역후，이용잡교류세포기술진행세포융합，경과다차아극륭사선출고민감적특이성항리춘혈증병독단극륭항체잡교류세포주，체내유생복수제비항리춘혈증병독단극륭항체병대기면역학특성진행감정。융합후사출량주잡교류세포주9C11화1C12，아형균위IgG2b형，κ련；간접ELISA측정복수효개위104，여기타병독주화세포주불발생반응；western-blot결과현시단극륭항체대응적항원위점재상대분자질량（Mr）230000적조대처；면역양화매염색결과현시단극륭항체능구특이지식별감염세포내적리춘혈증병독。본실험제비적항리춘혈증병독단극륭항체특이성호，위리춘혈증병독면역학쾌속검측방법적건립전정료기출。
The aim of the study is to generate high specificity monoclonal antibody against spring viraemia of carp virus (SVCV) and identify its immunological characteristics. The BALB/c mice were immunized with the SVCV purified by differential centrifugation. Spleen cells of immunized mice were collected and fused with SP2/0 myeloma cells. High sensitivity and high specificity monoclonal antibody was prepared after several times subcloning, and the immunological characteristics, characterized. Limited dilution method was used to subclone the positive clones. After three cycle of subcloning, two cell strains were obtained;it can secrete mAb against SVCV and were named as 9C11 and 1C12. The titer of mAbs ascites was 104 by the indirect ELISA. It belongs to IgG3 subclassκchain and couldn’t react with other fish viruses or fish cells lines in the ELISA test except SVCV. In the western blotting using mAbs (9C11 and 1C12), a 230 kDa of protein band was observed when SVCV was used as antigen. The result of the immunoperoxidase test showed that the monoclonal antibody can specifically recognize SVCV in infected cells. The obtained mAb could be used to rapidly diagnosis and detect SVCV in aquaculture.