Time-resolved immunofluorometry and other frequently used immunoassay types for follicle-stimulating hormone compared by using identical monoclonal antibodies.
The influence of assay design and quantification system on assay performance was investigated by developing, optimizing, and comparing a time-resolved immunofluorometric assay (IFMA), an immunoenzymometric assay (IEMA), an immunoradiometric assay (IRMA), and a competitive radioimmunoassay (RIA), all performed with the same monoclonal antibodies (MCA) directed against human follicle-stimulating hormone (hFSH). The lowest detection limit (2 ng/L for hFSH-I-3, corresponding to 2.5 mIU of 1st International Reference Preparation of hFSH 78/549 per liter), the widest measuring range (2-160,000 ng/L), and the greatest signal-to-noise ratio (13,000:1 at 160,000 ng/L) were obtained in the IFMA. For analysis of serum samples from 101 male (ages 2-91 years) and 99 female (ages 2-90 years) individuals at a single dilution, 100% of samples were within the measuring range of the IFMA, whereas only 87%, 55%, 32%, and 8% of the sera were for the IRMA, the IEMA evaluated with double-wavelength measurement, the conventional IEMA, and the competitive MCA-based RIA, respectively. These studies demonstrate clear advantages of the IFMA in sensitivity and assay range, which allows reliable and cost- and time-effective determination of hFSH in individuals from infancy to senescence.
Madersbacher
S
S
Institute for Biomedical Aging Research, Austrian Academy of Sciences, Innsbruck.
Shu-Chen
T
T
Schwarz
S
S
Dirnhofer
S
S
Wick
G
G
Berger
P
P
D000293:Adolescent; D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D000911:Antibodies, Monoclonal; D002648:Child; D002675:Child, Preschool; D005260:Female; D015200:Fluoroimmunoassay / Q000379:methods* / Q000592:standards / Q000706:statistics & numerical data; D005640:Follicle Stimulating Hormone / Q000097:blood*; D006801:Humans; D007118:Immunoassay / Q000379:methods* / Q000592:standards / Q000706:statistics & numerical data; D007124:Immunoenzyme Techniques / Q000592:standards / Q000706:statistics & numerical data; D015592:Immunoradiometric Assay / Q000592:standards / Q000706:statistics & numerical data; D008297:Male; D008875:Middle Aged; D011863:Radioimmunoassay / Q000592:standards / Q000706:statistics & numerical data; D012016:Reference Values
D000911:Antibodies, Monoclonal; D005640:Follicle Stimulating Hormone