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Clinical Chemistry 44: 1674-1679, 1998;
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(Clinical Chemistry. 1998;44:1674-1679.)
© 1998 American Association for Clinical Chemistry, Inc.

Automation and Analytical Techniques

Development of an enzyme-linked immunosorbent assay with monoclonal antibody for quantification of homovanillic in human urine samples

Run Zhang Shi1, Yee-Ping Ho1,a, John Hok Keung Yeung2, Penelope Mei Yu Or2, Kenneth Kin Wah To1, Mary Wai Man Lau1, and Mano Arumanayagam3

Departments of
1 Pharmacy,
2 Pharmacology, and
3 Chemical Pathology, Faculty of Medicine, The Chinese University of Hong Kong, Shatin, New Territories, Hong Kong.
a Author for correspondence. Fax 852-2603-5295; e-mail yeepingho{at}cuhk.edu.hk.

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.






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