Detection and quantification of secreted soluble Fc gamma RIIA in human sera by an enzyme-linked immunosorbent assay.
Fc gamma RIIA can be produced in a soluble form that contains both the extracellular and intracellular regions of the receptor, due to an alternative splicing of the transmembrane domain-coding exon. We have developed an enzyme-linked immunosorbent assay (ELISA) that permits the specific detection and quantification in human sera of this secreted soluble Fc gamma RIIA. It uses the monoclonal antibody (MAb) IV.3 as capture antibody and rabbit polyclonal IgG directed against the intracellular region of Fc gamma RIIA as detector antibodies. The enzymatic reaction was amplified using an NADH/NAD+ amplification system. As little as 0.8-1.5 ng/ml (20-38 pM) of purified recombinant secreted Fc gamma RIIA could be detected. The serum levels of secreted sFc gamma RIIA ranged from 0 to 30 ng/ml in sera from 51 healthy donors. The mean value was 11.9 ng/ml +/- 6.55 (297 pM +/- 163) and the median value was 10.6 ng/ml (265 pM) (range: 0-764 pM).
Astier
A
A
Unité INSERM 255, Institut Curie, Paris, France.
de la Salle
H
H
Moncuit
J
J
Freund
M
M
Cazenave
J P
JP
Fridman
W H
WH
Hanau
D
D
Teillaud
J L
JL
D000818:Animals; D000918:Antibody Specificity; D015703:Antigens, CD; D001483:Base Sequence; D016466:CHO Cells; D003001:Cloning, Molecular; D006224:Cricetinae; D018076:DNA, Complementary / Q000235:genetics; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D004926:Escherichia coli / Q000235:genetics; D006801:Humans; D007801:Langerhans Cells / Q000276:immunology; D008969:Molecular Sequence Data; D017452:Receptors, IgG / Q000032:analysis* / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000276:immunology; D012995:Solubility; D014162:Transfection
D015703:Antigens, CD; D018076:DNA, Complementary; C080568:Fc gamma receptor IIA; D017452:Receptors, IgG; D011993:Recombinant Fusion Proteins