Preparation Of Monoclonal Antibody To Matrix Protein Of Donkey Leucocyte Attenuated Equine Infectious Anemia Virus And Preliminary Identification Of Antigenic Epitopes

Six-week-old female Balb/c mice were immunized with purified recombinant DLV-MA protein.Seven hybridoma cell lines named 1F12,1E2,1E3,4B10,4H7,5E5,4G5 respectivly ,were obtained by using hybridoma technique, limiting dilution and three-time subclone.All the cell lines being screened by indirect ELISA,IFA and Western-Blot,secreted monoclonal antibody(McAb) specific against DLV-MA stably.The results showed that the McAb has high specificity to matrix protein of donkey leucocyte attenuated equine infectiousa nemia virus.The EIAV MA gene were divided into four overlapping fragments and expressed in E. coli respectively.The expressed fusion proteins were detected with sera of EIAV infected horse by Western-Blot.The results showed that there were four fragments named Peptide1,Peptide2,Peptide3 and Peptide4. The expressed fusion proteins and the McAb obtained were both analyzed by Western-Blot to identify epitopes of the McAb. The result was as following, the 1E3 McAb was matched by Peptide1,4H7 McAb was matched by Peptide2, and 4G5McAb was matched by Peptide3.The other four McAbs had no response with the peptides expressed which was considered as epitopes depended on conformation.Taken together,recombinant DLV-MA protein was used to prepare monoclonal antibodies in this study,and the epitope was identified by using the expressed protein from the segment gene.This may be helpful in understanding molecular properties of DLV-MA and may be useful for pathogenic and differential diagnosis.