Chemiluminescence sandwich enzyme immunoassay for determination of human granulocyte colony stimulating factor (G-CSF).
A chemiluminescence sandwich enzyme immunoassay, using a glucose oxidase (GO) label, was developed for detecting attomole amounts of human granulocyte colony stimulating factor (G-CSF). Purified goat (F(ab')2 immobilized on a bead and purified goat Fab' labelled with GO were selected in combination with a chemiluminescent detection system comprising luminol and ferricyanide. The detection limits for G-CSF were 4amol/assay (1 pg/mL) in buffer solution and 10 amol/assay (2.5pg/mL) in human serum. Coefficients of variation within assay and between assay ranged from 5.5% to 7.8% and from 3.4% to 16.0%, respectively. The C-CSF content of serum from normal healthy individuals was measurable using this method. G-CSF in 24 normal human sera showed a mean value of 19.3pg/mL and ranged from 3.6 to 83.0pg/mL.
Aoygai
S
S
R&D Center, Meidensha Co., Tokyo, Japan.
Arasawa
K
K
Matsuyuki
A
A
Kamatchi
S
S
Fukushima
M
M
Ohsawa
N
N
D000818:Animals; D005069:Evaluation Studies as Topic; D005292:Ferricyanides; D005949:Glucose Oxidase; D006041:Goats; D016179:Granulocyte Colony-Stimulating Factor / Q000032:analysis* / Q000097:blood / Q000276:immunology; D006801:Humans; D007124:Immunoenzyme Techniques / Q000706:statistics & numerical data; D007140:Immunoglobulin Fab Fragments; D008163:Luminescent Measurements; D008165:Luminol; D012016:Reference Values; D012680:Sensitivity and Specificity
D005292:Ferricyanides; D007140:Immunoglobulin Fab Fragments; C007931:hexacyanoferrate III; D016179:Granulocyte Colony-Stimulating Factor; D008165:Luminol; D005949:Glucose Oxidase