Recombinant antibody-alkaline phosphatase conjugates for diagnosis of human IgGs: application to anti-HBsAg detection.
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.
Carrier
A
A
Département d'Ingénierie et d'Etudes des Protéines, C.E.A. Saclay, Gif/Yvette, France.
Ducancel
F
F
Settiawan
N B
NB
Cattolico
L
L
Maillère
B
B
Léonetti
M
M
Drevet
P
P
Ménez
A
A
Boulain
J C
JC
D000469:Alkaline Phosphatase / Q000235:genetics; D000595:Amino Acid Sequence; D000911:Antibodies, Monoclonal / Q000276:immunology; D001483:Base Sequence; D004926:Escherichia coli; D006510:Hepatitis B Antibodies / Q000032:analysis*; D006514:Hepatitis B Surface Antigens / Q000276:immunology*; D006801:Humans; D007074:Immunoglobulin G / Q000032:analysis*; D008969:Molecular Sequence Data; D015345:Oligonucleotide Probes / Q000737:chemistry; D011993:Recombinant Fusion Proteins
D000911:Antibodies, Monoclonal; D006510:Hepatitis B Antibodies; D006514:Hepatitis B Surface Antigens; D007074:Immunoglobulin G; D015345:Oligonucleotide Probes; D011993:Recombinant Fusion Proteins; D000469:Alkaline Phosphatase