Development and testing of radio and enzyme immunoassays for acidic fibroblast growth factor (aFGF).
Acidic fibroblast growth factor (aFGF) and basic fibroblast growth factor from bovine brain stimulate growth in a variety of tissues in several species. Despite the 55% amino acid sequence homology of the two forms of FGF, a specific immunoassay of aFGF has been developed using a polyclonal antibody raised in a rabbit. Two immunoassays were compared: a radioimmunoassay (RIA) using 125I aFGF and an enzyme immunoassay (EIA) using aFGF coupled to the tetrameric form of acetylcholinesterase (aFGF-AchE) as tracer. With EIA, the detection limit was 1.5 ng/ml, versus 2.2 ng/ml with RIA, while the dose at 50% was 5.9 ng/ml for EIA and 9.6 ng/ml for RIA. Using a modified EIA procedure where aFGF-AchE was added 2 h after the other reagents, the dose at 50% binding was 1.5 ng/ml. Examples of the performance of both immunoassays are presented for various brain extracts of different species including human. The aFGF content obtained by these methods correlates (CR = 0.987) with the values obtained by biological assay.
Caruelle
D
D
Laboratoire de Biotechnologie des Cellules Eucaryotes, Universite Paris XII, Creteil, France.
Grassi
J
J
Courty
J
J
Groux-Muscatelli
B
B
Pradelles
P
P
Barritault
D
D
Caruelle
J P
JP
D000110:Acetylcholinesterase / Q000032:analysis / Q000302:isolation & purification; D000818:Animals; D000917:Antibody Formation; D000918:Antibody Specificity; D002417:Cattle; D002478:Cells, Cultured; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D005346:Fibroblast Growth Factors / Q000032:analysis*; D007114:Immunization; D015151:Immunoblotting; D007124:Immunoenzyme Techniques; D007457:Iodine Radioisotopes; D051379:Mice; D008934:Mitogens; D011817:Rabbits; D011863:Radioimmunoassay
D007457:Iodine Radioisotopes; D008934:Mitogens; D005346:Fibroblast Growth Factors; D000110:Acetylcholinesterase