Human brain lectin immunoreactive material in cerebrospinal fluids determined by enzyme immunoassay (EIA).
A sensitive enzyme immunoassay (EIA) micromethod is described which can measure levels of a 14 kDa human brain lectin (HBL) in the cerebrospinal fluid (CSF) of patients submitted to CSF examination. The assay is based on the use of a polyclonal antibody to HBL and the simultaneous application of biotinylated and unlabeled HBL. Biotin was then reacted with a streptavidin-peroxidase (Strep-HRP) conjugate and the bound enzyme quantified with the substrate orthophenylenediamine (OPD). The assay requires only 50 microliters of CSF and is very sensitive: as little as 6 ng/ml of HBL 14 can be detected. In a blind-test screening, the mean (+/- SEM) concentration of the HBL immunoreactive material (HIM) in CSF was determined to be 72.4 +/- 6.6 ng/ml. Our results indicate that EIA measurement of HIM levels in the CSF may find useful applications in elucidating the involvement of HBL in the physiopathology of human nervous system (NS).
Eloumami
H
H
Laboratoire de Biochimie et Technologie des Protéines, Université Paris-Nord, Bobigny, France.
Caron
M
M
Joubert
R
R
Doinel
C
C
Bladier
D
D
D000906:Antibodies; D015415:Biomarkers / Q000097:blood / Q000134:cerebrospinal fluid; D001921:Brain; D037161:Galectins; D006388:Hemagglutinins / Q000097:blood / Q000134:cerebrospinal fluid*; D006801:Humans; D007124:Immunoenzyme Techniques; D037102:Lectins / Q000097:blood / Q000134:cerebrospinal fluid*; D009422:Nervous System Diseases / Q000097:blood / Q000134:cerebrospinal fluid*
D000906:Antibodies; D015415:Biomarkers; D037161:Galectins; D006388:Hemagglutinins; D037102:Lectins