[Recombinant design and expression of human three-domain antibody against BoNTa].
VH and Vkappa genes were amplified from human ScFv B17 specific against botulinum neurotoxin serotype A (BoNTa). Gene sequence encoding 5'-terminal 12 amino acid of heavy chain constant region CH1, as a linker, linked VH and Vkappa to construct a new three-domain antibody molecule VH/Vkappa. VH/Vkappa was expressed at high level over 34% of total host cell proteins in E. coli. Recombinant protein were purified up to 95% by affinity column. As a result, recombinant VH/Vkappa could recognize and bind specific to BoNTa in ELISA. However, comparing with its parent ScFv, VH/Vkappa has higher relative affinity and stability.
Institute of Microbiology and Epidemiology, Academy of Military Medical Sciences, Beijing 100071, China. geno0109@vip.sina.com
Hui
H
Wang
Jun
J
Yin
Jing
J
Shi
Lu-lu
LL
Meng
Pei-zhen
PZ
Li
D000911:Antibodies, Monoclonal / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics / Q000276:immunology; D000918:Antibody Specificity; D019274:Botulinum Toxins, Type A / Q000276:immunology*; D004926:Escherichia coli / Q000235:genetics; D006801:Humans; D007128:Immunoglobulin Fragments / Q000737:chemistry / Q000235:genetics*; D007143:Immunoglobulin Heavy Chains / Q000737:chemistry / Q000235:genetics; D007147:Immunoglobulin Light Chains / Q000737:chemistry / Q000235:genetics; D007135:Immunoglobulin Variable Region / Q000737:chemistry / Q000235:genetics; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000737:chemistry / Q000276:immunology
D000911:Antibodies, Monoclonal; D007128:Immunoglobulin Fragments; D007143:Immunoglobulin Heavy Chains; D007147:Immunoglobulin Light Chains; D007135:Immunoglobulin Variable Region; D011994:Recombinant Proteins; C055966:immunoglobulin Fv; D019274:Botulinum Toxins, Type A