交叉中和血清1型和血清3型鸭甲肝病毒单克隆抗体的制备与鉴定

鸭甲型肝炎(duck hepatitis A,DHA)是危害养鸭业的主要疾病之一,是一种雏鸭的急性、高度致死性传染病.本研究旨在研制一种对我国流行的鸭甲型肝炎血清1型(DHAV-1)和血清3型(DHAV-3)病毒具有交叉中和作用的单克隆抗体(monoclonal antibody,McAb).人工合成12条DHAV-1...

Preparation and Identification of Monoclonal Antibodies with Cross-neutralizing Activity against Duck Hepatitis A Virus Serotype 1 and 3

Duck hepatitis A (DHA), an acute and highly lethal infection in ducklings, is one of the main diseases threatening the duck industry. The purpose of this study was to develop a monoclonal antibody (McAb) that has cross-neutralization activity against two most prevalent serotypes of DHAV in China, DHAV-1 and DHAV-3. Twelve epitope peptides shared by VP1 proteins of DHAV-1 and DHAV-3 were synthesized artificially and respectively conjugated with keyhole limpet hemocyanin (KLH) protein. BALB/c mice were then immunized with those antigens and McAbs were prepared. The obtained McAbs were screened by detecting the cross-reactivity with DHAV-1 and DHAV-3, the inhibitory efficiency on virus proliferation, the neutralization titer, and the protection rate against virus infection. A total of 12 hybridomas were obtained, the McAbs secreted by 6 hybridomas (C1, C4, C7, C12, C13, C16) in those had specific cross-reactions with DHAV-1 and DHAV-3. The McAbs secreted by hybridomas (C1, C4, C7 and C16) had higher inhibitory efficiency on DHAV-1 and DHAV-3 proliferation in duck embryos, ranging from 75.34% to 100.00%. The neutralization titer of McAbs secreted by hybridomas (C1, C4, C7 and C16) against DHAV-1 and DHAV-3 were 1:3 & 1:5, 1:3 & 1:3, 1:10 & 1:11, 1:9 & 1:9, respectively. The protection rate of McAbs secreted by hybridomas (C7 and C16) against DHAV-1 and DHAV-3 infection were 70% & 80%, 100% & 60%, respectively. In this study, four McAbs with cross-neutralizing activity against DHAV-1 and DHAV-3 were developed, two of those had a higher protection rate against virus infection, providing new materials and ideas for the prevention and control of DHA.