动物医学进展
動物醫學進展
동물의학진전
PROGRESS IN VETERINARY MEDICINE
2014年
3期
73-77
,共5页
王铮%王铁成%冯娜%虞一聪%高玉伟%杨松涛%夏咸柱
王錚%王鐵成%馮娜%虞一聰%高玉偉%楊鬆濤%夏鹹柱
왕쟁%왕철성%풍나%우일총%고옥위%양송도%하함주
犬瘟热病毒%单克隆抗体%间接酶联免疫吸附试验%相加酶联免疫吸附试验
犬瘟熱病毒%單剋隆抗體%間接酶聯免疫吸附試驗%相加酶聯免疫吸附試驗
견온열병독%단극륭항체%간접매련면역흡부시험%상가매련면역흡부시험
Canine distemper virus%monoclonal antibody%indirect ELISA%additivity ELISA
用纯化后的犬瘟热病毒免疫Balb/c小鼠,采用间接ELISA方法筛选分泌阳性抗体的杂交瘤细胞株,并挑选阳性孔进行亚克隆,用Vero细胞交叉反应试验及间接免疫荧光检测单克隆抗体的特异性。结果获得2株能稳定分泌抗犬瘟热病毒单克隆抗体细胞株,分别命名为062-1、062-2。实验室已有的一株单克隆抗体为6 H8。交叉反应试验及间接免疫荧光染色检测表明,3株单克隆抗体的特异性较好;经过体外连续传代及反复冻存、复苏,杂交瘤细胞均能稳定分泌特异性单克隆抗体。培养上清液及小鼠腹水间接 ELISA效价分别为1∶105和1∶108。相加试验表明,单克隆抗体062-1、062-2具有共同的表位,而单克隆抗体6 H8识别的位点与其他两株不同。
用純化後的犬瘟熱病毒免疫Balb/c小鼠,採用間接ELISA方法篩選分泌暘性抗體的雜交瘤細胞株,併挑選暘性孔進行亞剋隆,用Vero細胞交扠反應試驗及間接免疫熒光檢測單剋隆抗體的特異性。結果穫得2株能穩定分泌抗犬瘟熱病毒單剋隆抗體細胞株,分彆命名為062-1、062-2。實驗室已有的一株單剋隆抗體為6 H8。交扠反應試驗及間接免疫熒光染色檢測錶明,3株單剋隆抗體的特異性較好;經過體外連續傳代及反複凍存、複囌,雜交瘤細胞均能穩定分泌特異性單剋隆抗體。培養上清液及小鼠腹水間接 ELISA效價分彆為1∶105和1∶108。相加試驗錶明,單剋隆抗體062-1、062-2具有共同的錶位,而單剋隆抗體6 H8識彆的位點與其他兩株不同。
용순화후적견온열병독면역Balb/c소서,채용간접ELISA방법사선분비양성항체적잡교류세포주,병도선양성공진행아극륭,용Vero세포교차반응시험급간접면역형광검측단극륭항체적특이성。결과획득2주능은정분비항견온열병독단극륭항체세포주,분별명명위062-1、062-2。실험실이유적일주단극륭항체위6 H8。교차반응시험급간접면역형광염색검측표명,3주단극륭항체적특이성교호;경과체외련속전대급반복동존、복소,잡교류세포균능은정분비특이성단극륭항체。배양상청액급소서복수간접 ELISA효개분별위1∶105화1∶108。상가시험표명,단극륭항체062-1、062-2구유공동적표위,이단극륭항체6 H8식별적위점여기타량주불동。
Canine distemper virus was purified and used to immunize Balb/c mice.The hybridoma culture su-pernants were screened by indirect ELISA.The positive cells were subcloned until all clones were positive. The specificity of monoclonal antibodies was confirmed by Vero cross-reaction test and IFA.The results showed that three hybridoma cells secreting anti-CDV McAb were obtained.These two cells were named 062-1,062-2.The lab had one McAb called 6H8.All the three McAbs confirmed by cross-reaction test and IFA were specific against CDV.The positive hybridomas were also able to secrete McAbs stably by series passage in vitro with freezing and recovering.The titers of cell culture supernant and ascites tested by ELISA were 1∶105 and 1∶108 ,respectively.Additivity ELISA results showed that epitopes recognized by 2 McAbs(062-1,062-2)differed from that of by McAb 6H8.