Construction of a phage display library of human single-chain Fv antibodies from peripheral blood lymphocytes of people with positive serum antibody against hepatitis B virus core protein
- VernacularTitle:人源性抗-HBc单链噬菌体抗体库的构建
- Author:
Zhenghao TANG
;
Huihui MA
;
Gang LI
- Publication Type:Journal Article
- Keywords:
Antibodies;
Bacteriophages;
Phage surface display technology;
Polymerase chain reaction;
Hepatitis B core antigers
- From:
Chinese Journal of Infectious Diseases
2001;0(06):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective To construcr a phage display library of human single-chain Fv antibodies from peripheral blood lymphocytes of people with positive serum antibody against hepatitis B virus core protein. Methods The heavy chain and light chain variable region gene of human immunoglobulin were amplified individually by RT-PCR from peripheral blood Lymphocytes mRNA of patients with positive serum hepatitis B virus core protein antibody and randomly combined through a DNA linker encoding the peptide (Gly 4Ser) 3 to construct single-chain variable fragments (ScFv)gene. In the presence of helper phage M13KO7, the ScFv were displayed on the surface of recombinant phages. Results Repertoire antibody library was obtained with 10 6 clonies resistant to ampicillin and kanamycin. Conclusions A typical phage display library of human single-chain Fv antibodies has been constructed Using RT-polymerase chain reaction (RT-PCR) and phage display technique, human single-chain Fv antibodies can be screened from this library.
