Abstract
Although considerable evidence supports the role of EGF as a modulator of developmental processes, data from adult humans are limited due to the lack of reliable measurement methods. There is no information from infants or children. We developed and characterized a homologous RIA system to implement clinical hEGF studies. Antibodies were raised in NZW rabbits against highly purified Chiron h-EGF. The same preparation was used as standard and radioiodinated with chloramine T. The antibody (1.5×105 final dilution) was incubated for 48 h at 7°C with standard and samples. Radiolabeled hEGF was added at 24 h and bound-free separation was performed by combined 2nd antibody and 5% polyethylene glycol. Sensitivity of the method is 30 pg; there is no crossreaction with insulin, placental NGF extract, IGF-I, IGF-II or renin up to levels of 100 ng. Within and across assay variations were 7.9 and 12.3%, respectively. Human EGF measured in urine of term newborns expressed in ng/mg of creatinine were 9.9±3.6 (x±SD) at 0 h and 3.9±1.8 at 24 h after birth (p < 0.05). These values were lower than levels in normal boys and girls 6-8 yr of age (68±16, p < 0.01) or normal men (48±13, p < 0.01). In cord serum hEGF was 0.62±0.36 ng/ml.
Conclusion: The present hEGF RIA system provides a reliable and sensitive measurement method which offers an unique possibility for the study of pathophysiological implications of EGF in human development.
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Callegari, C., Laborde, N., Lakshmanan, J. et al. 91 A HIGHLY SENSITIVE RIA SYSTEM FOR HUMAN EPIDERMAL GROWTH FACTOR (EGF). Pediatr Res 19, 618 (1985). https://doi.org/10.1203/00006450-198506000-00111
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DOI: https://doi.org/10.1203/00006450-198506000-00111