[Recombinant colorimetric antibodies: genetic construction and production in E. coli].
We constructed an expression vector comprising a transcription unit composed of (1) the gene of alkaline phosphatase from E. coli in which we inserted a DNA fragment encoding the VH and CH domains of an IgG2A; (2) a DNA fragment encoding the light chain of the same immunoglobulin. Bacteria, E. coli were transformed with the plasmid, and hence it produced a periplasmic and chimaeric protein having both the alkaline phosphatase enzymatic activity and the immunoglobulin binding function. The hybrid protein mimics a bivalent antibody whose Fc part is replaced by a dimer of alkaline phosphatase. Recombinant colorimetric antibodies offer interesting potentialities for future developments in the field of immuno-enzymatic diagnosis.
Ducancel
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F
Département d'Ingénierie et d'Etudes des Protéines, C.E.A. Saclay, Gif-sur-Yvette.
Gillet
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D
Ménez
A
A
Boulain
J C
JC
D000469:Alkaline Phosphatase / Q000235:genetics; D000907:Antibodies, Bacterial / Q000235:genetics*; D003124:Colorimetry; D004251:DNA Transposable Elements / Q000235:genetics; D004926:Escherichia coli / Q000201:enzymology / Q000276:immunology*; D005822:Genetic Vectors / Q000235:genetics; D007140:Immunoglobulin Fab Fragments / Q000235:genetics; D010957:Plasmids / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000235:genetics*; D011995:Recombination, Genetic
D000907:Antibodies, Bacterial; D004251:DNA Transposable Elements; D007140:Immunoglobulin Fab Fragments; D011993:Recombinant Fusion Proteins; D000469:Alkaline Phosphatase
Anticorps colorimétriques recombinants: construction génétique et production chez E. coli.