Related Experiment Videos
[A three-domain antibody fragment VH/L specific to tumor blood vessels]
1State Key Laboratory of Microbial Resource, Institute of Microbiology, Chinese Academy of Sciences, Beijing 100080, China.
AA98 is a specificaally anti-angiogenic antibody generated in our lab. The heavy chain variable region (VH) attached with mutagenized 36 nucleotides sequence derived from the heavy chain constant region1 (CH1) was amplified VH and light chain (L) were inserted into high-level expression vector pET21a successively, thus pET21a-VH/L was constructed. VH/L was expressed in E. coli BL21 (DE3) after induction with IPTG. The expression of VH/L was 20% of the total bacterial proteins. The refolding of VH/L was conducted by dilution and gel filtration chromatography. The refolded VH/L could bind to HUVEC specifically. Its affinity to the antigen is similar to that of recombinant AA98Fab, but lower than that of the parent antibody AA98.
AA98 is a specificaally anti-angiogenic antibody generated in our lab. The heavy chain variable region (VH) attached with mutagenized 36 nucleotides sequence derived from the heavy chain constant region1 (CH1) was amplified VH and light chain (L) were inserted into high-level expression vector pET21a successively, thus pET21a-VH/L was constructed. VH/L was expressed in E. coli BL21 (DE3) after induction with IPTG. The expression of VH/L was 20% of the total bacterial proteins. The refolding of VH/L was conducted by dilution and gel filtration chromatography. The refolded VH/L could bind to HUVEC specifically. Its affinity to the antigen is similar to that of recombinant AA98Fab, but lower than that of the parent antibody AA98.