Preparation of recombinant anti-tau antibody and its application in the prevention and treatment of Alzheimer's disease

HUANG Ning; WANG Shengping; WU Yanhong; YU Qinghua; GUO Weiwei

doi:10.3969/j.issn.2096-9058.2025.04.006

Abstract：Objective To investigate the preparation of recombinant anti-tau antibody and its application in the pre-vention and treatment of Alzheimer's disease(AD).Methods Recombinant human tau441 protein was used as immuno-gen to obtain mouse lymphocytes secreting anti-tau antibody.The mouse immune VH-VL gene bank was constructed,and the correctly sequenced positive recombinant plasmid was transformed into PCDNA3.1(+).After the recombinant anti-tau antibody was purified by affinity chromatography,its titer,purity and specificity were identified by ELISA,gel electropho-resis and Western blotting.The mouse brain neuroma N2a cells were used as the research object.The N2a cells were divid-ed into a blank control group(normal culture),an AD group(establishing an AD cell model by inducing with Aβ25-35),and an recombinant anti-tau antibody group(treated with anti-tau antibody after establishing the AD cell model by inducing with Aβ25-35).CCK-8 assay was used to detect cell viability,and Hoechst 33342 staining was used to analyze cell apopto-sis.Results Mice were successfully immunized and anti-tau antibody VH and VL genes were obtained.Through the con-struction of VH-linker-VL vector and purification by affinity chromatography,specific recombinant anti-tau antibody with titer higher than 1∶256 000 and purity of 95.28%was obtained.The recombinant anti-tau antibodies can enhance the via-bility of N2a cells induced by Aβ25-35,and inhibit their apoptosis.The cell viability of the AD group and the recombinant an-ti-tau antibody group was(44.33±2.08)%and(68.00±2.65)%respectively.There was a statistically significant dif-ference between the two groups(P<0.05).Conclusion In this study,the VH and VL sequences of anti-tau antibody are obtained and the recombinant anti-tau antibody with high titer,high purity and high specificity are prepared,and its protective effect on N2a cell injury by inducing with Aβ25-35 is confirmed.