摘要
以灭活的H9亚型禽流感NJ01病毒株作为抗原，按常规方法免疫BALB／c鼠，最后一次免疫后第3d取脾细胞与SP2／0细胞进行融合，采用HI进行筛选，检出18个阳性孔，阳性率为4．69％。对其中的3个阳性值较高的孔进行3次克隆，最终获得3株单克隆细胞，分别命名为1E1、2E10和3G2。通过与新城疫病毒、产蛋下降综合征病毒、H5亚型禽流感病毒进行HI试验，证明1E1、2E10和3G2分泌的抗体具有特异性。经连续1个月的体外培养和两次冻存复苏仍能稳定地分泌抗H9亚型禽流感Hl抗体，3株杂交瘤细胞分泌的单克隆抗体均为IgG1，杂交瘤细胞的染色体数目为92—108条，平均97条。杂交瘤细胞培养上清和腹水对H9亚型禽流感病毒的Hl值分别为2^5～2^8和2^12～2^14.PD50分别10^-2～10^-1.67和10^-4～10^-3.33单克隆抗体的制备为禽流感的快速诊断、预警预报和病毒的抗原性分析等奠定了基础。
The monoclonal antibody（McAb） specific to the H9 subtype of avian influenza virus （AIV） was developed by fusing SP2/0 myeloma cell with spleen cells of mice immunized with inactivated AIV-H9（ NJ01 strain） subtype vaccine, and positive hybridoma clones were screened by hemagglutination inhibition test （HI）. Eighteen positive reactions up to 4.69 per cent were screened by HI. Three of them were cloned three times and the clones were designated as 1E1,2El0, and 3G2 respectively, which were specific to AIV-Hg. AIV-H9 antibodies were secreted steadily in one-month in vitro culture and twice refrigerated storage and revivification. The three McAbs were identified as IgG1, and the chromosome number of hybridoma cell were 93-108, 97 average. The HI titers of supernatants of hybridoma cell culture and ascites reacted with AIV-H9 were 2^5-2^8 and 2^12-2^14, and the PD50 were 10^-2～10^-1.67 and 10^-4～10^-3.33. The development of three McAbs may provide an useful tool in the fast diagnosis and the forecast of AIV.
出处
《江苏农业学报》
CSCD
北大核心
2006年第2期127-130,共4页
Jiangsu Journal of Agricultural Sciences
基金
江苏省科技成果转化专项资金"兽用疫苗成果转化与产业化示范"(BA2004032)
关键词
H9亚型禽流感病毒
单克隆抗体
HI
生物学特性
H9 subtype of avian influenza virus
monoclonal antibody
hemagglutination inhibition test
biological characteristic
