Recombinant anti-stefin A Fab fragment: sequence analysis of the variable region and expression in Escherichia coli.
Biol Chem
; 381(12): 1245-9, 2000 Dec.
Article
en En
| MEDLINE
| ID: mdl-11209759
ABSTRACT
Human stefin A is an inhibitor of lysosomal cysteine proteinases cathepsin B, H, L and S. In the present report we describe the cloning and expression of anti-stefin A Fab fragment A22 in E. coli. We have determined the nucleotide sequences of the antibody heavy and light chain and compared them to the murine immunoglobulin germ line sequences. Expression of the two antibody chains was achieved using a single vector with a PhoA promoter and coding regions placed after the signal sequences, directing them to the periplasmic space. The A22 Fab fragment was extracted from the periplasmic space and expression was confirmed by Western blot analysis. The recombinant A22 Fab fragment had an affinity for stefin A comparable to the original monoclonal antibody, as determined by ELISA.
Buscar en Google
Base de datos:
MEDLINE
Asunto principal:
Fragmentos Fab de Inmunoglobulinas
/
Cistatinas
Límite:
Humans
Idioma:
En
Revista:
Biol Chem
Asunto de la revista:
BIOQUIMICA
Año:
2000
Tipo del documento:
Article
País de afiliación:
Eslovenia