Production of Human Fab Monoclonal Antibody to Surface Protein, preS1, of Hepatitis B Virus using Antibody Phage Display Library.
- Author:
Hee Sun KIM
;
In Hak CHOI
;
Ik Jung KIM
;
Jun Ho CHUNG
;
Se Kwang PARK
- Publication Type:Original Article
- Keywords:
Human Fab monoclonal antibody;
Hepatitis B virus;
Antibody phage display library
- MeSH:
Antibodies;
Antibodies, Monoclonal;
B-Lymphocytes;
Bacteriophages*;
Clone Cells;
Cloning, Organism;
Enzyme-Linked Immunosorbent Assay;
Genes, Immunoglobulin;
Hepatitis B virus*;
Hepatitis B*;
Hepatitis*;
Humans*;
Immunoglobulin Fab Fragments;
Isopropyl Thiogalactoside;
Streptavidin;
Virus Diseases;
Volunteers
- From:Korean Journal of Immunology
1997;19(4):549-558
- CountryRepublic of Korea
- Language:Korean
-
Abstract:
Human monoclonal antibodies have considerable potential in the prophylaxis and treatment of viral disease. By cloning human Ig gene segments from the B cells of volunteer into pComb3 phagemid vector, antibody library was created of filamentous phage particles displaying Fab fragments on their surface after being rescued with M13KO7 helper phages. The size of library was 7x10' pfu. Phage antibodies (phabs) were panned against biotinylated preS1 using streptavidine coated Dynabead. The soluble Fab antibodies were prepared from phagemid colonies and assayed directly for the ability to bind preS1 by ELISA. And then 3DW and SGW specific to preS1 which have both heavy and light chain to form Fab fragment, were selected. The soluble Fab antibody from 3DW was expressed highly at the concentration of 0.1 - 1.0 mM of IPTG, and 5 hours postinduction. The soluble antibodies from 3DW and SGW showed their relative affinities of 2x10' M ', and Sx10 M ', respectively, and the specificities to preS1 on ELISA. Our results suggest that antibody phage display library is very useful method to generate the human monoclonal antibody and that the human Fab monoclonal antibodies specific to preS1 selected in this study open the way to treat hepatitis B as a component of passive irnmunotherapeutics.
