Recombinant technology in the preparation of immunogen and enzymatic tracer. Application to the development of an enzyme immunoassay for rat prolactin.
A competitive enzyme immunoassay for rat prolactin using an immunogen and a tracer obtained by recombinant DNA technology is described. Polyclonal antibodies were raised in rabbits immunized with a purified chimeric protein consisting of rat prolactin fused with two synthetic immunoglobulin G binding domains derived from staphylococcal Protein A. The enzymatic tracer was obtained using an expression system which permits insertion of rPRL DNA sequence in the alkaline phosphatase gene. Antibodies and tracer were used to develop a solid-phase competitive immunoassay for the measurement of rat prolactin in plasma with a minimal detectable concentration of 0.5 ng/ml. The mean intra- and inter-assay coefficients of variation were 7.8 and 13.2%, respectively. Rat plasma concentrations measured with this assay correlated well with those obtained with a conventional enzyme immunoassay (r = 0.993, slope = 1.037, n = 24).
Ezan
E
E
Service de Pharmacologie et d'Immunologie, DRIPP Bât 136, Commissariat à l'Energie Atomique, CE/Saclay, Gif-sur-Yvette, France.
Ducancel
F
F
Gillet
D
D
Drevet
P
P
Ménez
A
A
Grognet
J M
JM
Boulain
J C
JC
D000469:Alkaline Phosphatase / Q000235:genetics*; D000818:Animals; D001483:Base Sequence; D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression; D007124:Immunoenzyme Techniques; D007074:Immunoglobulin G / Q000235:genetics / Q000276:immunology; D008297:Male; D008969:Molecular Sequence Data; D010957:Plasmids; D011388:Prolactin / Q000032:analysis* / Q000235:genetics; D011817:Rabbits; D051381:Rats; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000276:immunology; D011994:Recombinant Proteins / Q000032:analysis / Q000235:genetics
D007074:Immunoglobulin G; D011993:Recombinant Fusion Proteins; D011994:Recombinant Proteins; D011388:Prolactin; D000469:Alkaline Phosphatase