[Construction and expression in E. coli of single-chain Fv fragment of antihuman activated platelets monoclonal antibody].
OBJECTIVE
In order to reduce the immunogenicity of murine monoclonal antibody (MAb) to human beings.


METHODS
The cDNA encoding the variable regions of the mouse MAb SZ-51, which is specific against alpha-granule membrane protein(GMP) 140 on activated human platelets, was attached to the oligonucleotide of the linker peptide (Gly4 Ser)3 by means of recombinant DNA technique. The phagemid construct pHEN1-51scFv was obtained and introduced into the non-suppressor E. coli strain HB2151 for expression.


CONCLUSIONS
The expressed product could fold up into a soluble single-chain Fv fragment (scFv) and was released into the conditioned medium. The SZ-51scFv was proved to bind specifically to GMP140 by Western blot.
Wan
H
H
Jiangsu Institute of Hematology, First Affiliated Hospital, Suzhou Medical College, Suzhou 215007.
Liu
Y
Y
Gu
J
J
D000818:Animals; D000911:Antibodies, Monoclonal / Q000097:blood / Q000737:chemistry / Q000235:genetics* / Q000276:immunology*; D000918:Antibody Specificity; D001792:Blood Platelets; D004926:Escherichia coli / Q000235:genetics*; D015870:Gene Expression; D006801:Humans; D051379:Mice; D019007:P-Selectin / Q000276:immunology; D010957:Plasmids / Q000235:genetics; D057127:Single-Chain Antibodies / Q000097:blood / Q000737:chemistry / Q000235:genetics* / Q000276:immunology*; D012995:Solubility
D000911:Antibodies, Monoclonal; D019007:P-Selectin; D057127:Single-Chain Antibodies