Monoclonal Antibody Anti-Adeno-associate | 252-03-61058

A series of monoclonal antibodies against adeno-associated virus type 2(AAV2) were generated and their biological characteristics were analyzed briefly.

The purified recombinant AAV2 particles containing GFP gene (rAAV2-GFP) were used for intra-abdominal immunization and two boost injections and the plasma were screened by AAV2 particles ELISA.

The rAAV2-GFP was generated by infecting BHK-21 cells carrying pSNAV-GFP plasmids with rHSV1-rc, a recombinant herpes simplex virus type 1 carrying AAV2 rep and cap gene we produced previously.

Polyethylene glycol precipitation and chloroform extraction were used in purification pretreatment followed by cation exchange and molecular-size chromatography. The purity of rAAV2-GFP was more than 98% analyzed by HPLC and SDFS-PAGE.

Seven MAbs were isolated. Five of them were characterized as neutralizing antibodies (B10,B7,E1,H6,G4), with one of them(G4) showing strong inhibition of virus attachment to cells.

Subtype assay demonstrates that one is IgG1 (B10), two are IgM (B7 and G10), two IgG3 (B9 and H6),one IgG2a(G4) and one IgG2b(E1).

Western blotting assay demonstrated that all these seven antibodies recognize VP1,VP2 and VP3 of AAV2, suggested that the recognized epitopes were all resided in VP3. Further experiments need to be done to give more informations about these antibodies.