Cloning and Expressing of An Anti-CD5 Single Chain Antibody
抗CD5单链抗体基因克隆和表达

Novel anti\|CD5 single\|chain fragment of variable domain (ScFv) was cloned and expressed.Anti\|CD5 ScFv was constructed with cDNA fragments of heavy and light variable regions (V H and V L) which were reverse\|translated from poly (A) mRNA of hybridoma cells producing anti\|CD5 McAb.By phage displaying,ScFv in form of ScFv\|g3p fusion protein was panned based on its binding capacity to the CD5 antigens on the cell surface of Molt\|4 cells.Assayed by the cell\|ELISA,4 clones were found to have high affinity to CD5 antigen.DNA sequencing confirmed that the V H fragment is 339 base pairs,the V L 300bp,and the ScFv belongs to the mice gene family.In \%E.coli\% HB2151 the soluble ScFv\|CD5 was expressed mainly in the periplasm.