Development of an enzyme-linked immunosorbent assay with monoclonal antibody for quantification of homovanillic acid [corrected] in human urine samples.
A monoclonal antibody to homovanillic acid (HVA) was prepared by synthesis of a HVA-protein conjugate (HVA-ovalbumin) as an immunogen, immunization of mice, and the subsequent hybridization technique. Monoclonal antibodies were screened on the basis of sensitivity, specificity, and accuracy. An indirect ELISA was developed for quantification of HVA in human urine. The assay was characterized and shown to have high specificity, with cross-reactivities to vanillylmandelic acid and normetanephrine at 0.18% and <0.1%, respectively. The assay coefficients of variation were <10% within the working range of 0.5-40 mg/L. Initial results from testing urine samples of patients with neuroblastoma and other diseases were validated by HPLC, suggesting that this ELISA method is a reliable and convenient system for quantification of HVA in urine and can be used in the mass screening of neuroblastoma in infants.
Shi
R Z
RZ
Department of Pharmacy, Faculty of Medicine, The Chinese University of Hong Kong, Shatin, New Territories.
Ho
Y P
YP
Yeung
J H
JH
Or
P M
PM
To
K K
KK
Lau
M W
MW
Arumanayagam
M
M
D000293:Adolescent; D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis / Q000276:immunology; D002648:Child; D002675:Child, Preschool; D002851:Chromatography, High Pressure Liquid; D003429:Cross Reactions; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes; D006433:Hemocyanins / Q000276:immunology; D006719:Homovanillic Acid / Q000276:immunology / Q000652:urine*; D006801:Humans; D007114:Immunization; D007223:Infant; D007231:Infant, Newborn; D008297:Male; D051379:Mice; D009447:Neuroblastoma / Q000652:urine; D010047:Ovalbumin / Q000276:immunology; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
D000911:Antibodies, Monoclonal; D000939:Epitopes; D010047:Ovalbumin; D006433:Hemocyanins; C032808:keyhole-limpet hemocyanin; D006719:Homovanillic Acid