Abstract: Objective To produce and purify recombinant adeno-associated virus (AAV) loaded with anti-amyloid β peptide single-chain antibody gene for Alzheimer's disease gene therapy. Methods The plasmid pSNAV2.0-Abeta-scFv was utilized to transform BHK-21 cell by Lipofectamine 2000 for stable package cell line establishment. The helper virus HSV1-rc/ UL2 was used to transfect package cell for anti-amyloid β peptide single-chain antibody gene loaded recombinant adeno-associated virus production. The chloroform-PEG/NaCl-chloroform extraction and ion-exchange chromatography were employed for recombinant AAV purification. SDS-PAGE and PCR amplification were adopted for purified virus identification. The final virus physical titer was determined by digoxin-labeled DNA probes. The effect of gene therapy was tested with transgenic mice by Water maze test. Results The purity of recombinant adeno-associated virus with our target gene reach up to 98% after stable cell package and serial purification. The physical titer of the final virus was 1×1012vg/ml. The latency of treated mice in water maze test were reduced significantly. Conclusion The adeno-associated virus carrying anti-amyloidβ peptide single-chain antibody gene was produced by HSV1system and purified. The animal behavior test demonstrated the recombinant AAV was effective in Alzheimer's disease gene therapy.