Recombinant antibody-alkaline phosphatase conjugates for diagnosis of human IgGs: application to anti-HBsAg detection

A Carrier; F Ducancel; N B Settiawan; L Cattolico; B Maillère; M Léonetti; P Drevet; A Ménez; J C Boulain

doi:10.1016/0022-1759(94)00344-v

Recombinant antibody-alkaline phosphatase conjugates for diagnosis of human IgGs: application to anti-HBsAg detection

J Immunol Methods. 1995 Apr 26;181(2):177-86. doi: 10.1016/0022-1759(94)00344-v.

Authors

A Carrier¹, F Ducancel, N B Settiawan, L Cattolico, B Maillère, M Léonetti, P Drevet, A Ménez, J C Boulain

Affiliation

¹ Département d'Ingénierie et d'Etudes des Protéines, C.E.A. Saclay, Gif/Yvette, France.

PMID: 7745247
DOI: 10.1016/0022-1759(94)00344-v

Abstract

We have designed an expression vector permitting the production in the periplasm of Escherichia coli of a fusion protein comprising a dimer of bacterial alkaline phosphatase (PhoA) and two Fab or scFv fragments of a monoclonal antibody directed against human IgG. Each hybrid protein expressed both high specificity for the antigen and full PhoA activity. We show that crude periplasmic extracts containing these conjugates can be used as such in enzyme immunoassays for the detection of human IgG, as exemplified in the case of anti-hepatitis B immunoglobulin.

MeSH terms

Alkaline Phosphatase* / genetics
Amino Acid Sequence
Antibodies, Monoclonal / immunology
Base Sequence
Escherichia coli
Hepatitis B Antibodies / analysis*
Hepatitis B Surface Antigens / immunology*
Humans
Immunoglobulin G / analysis*
Molecular Sequence Data
Oligonucleotide Probes / chemistry
Recombinant Fusion Proteins

Substances

Antibodies, Monoclonal
Hepatitis B Antibodies
Hepatitis B Surface Antigens
Immunoglobulin G
Oligonucleotide Probes
Recombinant Fusion Proteins
Alkaline Phosphatase

Associated data

GENBANK/S77425
GENBANK/S77453