Expression, Characterization and Chain Shuffling of an Anti-HBsAg Phage Antibody.
- Author:
In Hak CHOI
;
Ik Jung KIM
;
Suk Jin CHOI
;
Hyo Jung KIM
;
Jun Ho CHUNG
;
Kye Sook YI
;
Sang Do LEE
;
Pann Ghill SUH
- Publication Type:Original Article
- MeSH:
Bacteriophages*;
Base Sequence;
Blotting, Western;
Clone Cells;
Hepatitis B Surface Antigens;
Humans;
Immunoglobulin G;
Periplasm;
Sensitivity and Specificity
- From:Journal of the Korean Society for Microbiology
1998;33(3):307-316
- CountryRepublic of Korea
- Language:Korean
-
Abstract:
In our previous report an anti-HBsAg human monoclonal antibody was generated using antibody phage display library technique. Using pComb3 filamentous phagemid vector, Fab molecule was expressed in fusion form to a phage coat protein in the periplasm of E. coli. A clone of HBsAg binder was selected after panning and designated as B7. In order to select the clones with higher affinity and to examine which chain contributes most to the affinity of B7, the light and heavy chain of B7 was sequentially deleted and replaced with new library. HBsAg-binders were selected and tested by EIA (enzyme immunoassay). It was revealed that the affinity of B7 depends only on the heavy chain of Fd. B7 Fd was constructed without light chain and specificity and affinity was further confirmed by western blot analysis. This human monoclonal Fd antibody was found to react with d antigenic determinant of HBsAg as the original clone did. The nucleotide sequence analysis revealed that VH of B7 could be classified into Kabat's subgroup II and human IgG heavy chain family IV. The CH1 of B7 was IgG1.
