Sensitive time-resolved fluorescence immunoassay of somatotropin in serum.
We describe a new "sandwich"-type non-isotopic immunoassay for human somatotropin (GH, growth hormone) in serum. In the assay, GH is captured by a monoclonal antibody immobilized in a white microtiter well and simultaneously reacted with a second biotinylated monoclonal antibody. The degree of binding of biotinylated antibody, which increases with increasing amount of GH in the sample, is quantified by adding streptavidin labeled with the europium chelate of 4.7 - bis(chlorosulfophenyl) - 1.10 - phenanthroline - 2.9 - dicarboxylic acid. The fluorescent complex on the solid phase is then measured by excitation at 337.1 nm (nitrogen laser) and monitoring the emission at 615 nm in a gated fluorometer/analyzer. The proposed procedure has short incubation times (less than 4 h protocol), uses only 25 microL of serum per microtiter well, and gives precise and accurate results. The method was clinically evaluated with samples obtained from pediatric patients undergoing investigation for growth abnormalities and from a patient with acromegaly.
Kahan
I
I
CyberFluor Inc., Toronto, Canada.
Papanastasiou-Diamandi
A
A
Ellis
G
G
Makela
S K
SK
McLaurin
J
J
D'Costa
M
M
Diamandis
E P
EP
D000172:Acromegaly / Q000097:blood; D000328:Adult; D000911:Antibodies, Monoclonal; D001426:Bacterial Proteins; D001710:Biotin; D005063:Europium; D005456:Fluorescent Dyes; D005470:Fluorometry; D013006:Growth Hormone / Q000097:blood*; D006801:Humans; D007118:Immunoassay / Q000706:statistics & numerical data; D010618:Phenanthrolines; D011863:Radioimmunoassay; D012016:Reference Values; D019809:Streptavidin
D000911:Antibodies, Monoclonal; D001426:Bacterial Proteins; D005456:Fluorescent Dyes; D010618:Phenanthrolines; C054206:4,7-bis(chlorosulfophenyl)-1,10-phenanthroline-2,9-dicarboxylic acid; D005063:Europium; D001710:Biotin; D013006:Growth Hormone; D019809:Streptavidin