Immunoreactive Somatomedin-C/Insulin-Like Growth Factor I and Its Binding Protein in Human Milk*

Abstract

Milk contains cell growth factors, but somatomedin-C/insulin-like growth factor I has not previously been identified. Acid-ethanol extracts of fresh human milk samples displaced somatomedin-C tracer from the specific somatomedin-C antiserum Tr4 parallel to standard. In samples obtained between days 1 and 9 postpartum, highest levels [17.6 +/- 10.4 (+/- SD) ng/ml; n = 7) were found in day 1 samples; these fell to stable levels of 6-8 ng/ml over the next several days. Sephadex G-200 chromatography of fresh 1- to 2-day-postpartum milk samples revealed immunoreactive peaks of approximately 150,000 and 40,000 mol wt, accounting for about 80% of the immunoreactivity; the remainder corresponded in elution volume to free somatomedin-C. On chromatography of the high molecular weight fractions in 1 M acetic acid, most of the immunoreactivity shifted to the position of somatomedin-C. Milk samples stripped of their endogenous somatomedins by ion exchange chromatography at low pH specifically bound somatomedin-C tracer in proportion to the amount of added protein up to 20% binding. Displacement of tracer by unlabeled somatomedin-C indicated an association constant of about 1 X 10(10) liter/mol. Fresh milk incubated with somatomedin-C tracer and fractionated by neutral gel chromatography bound the tracer predominantly in the 40,000 mol wt region, which was abolished when excess unlabeled peptide was included in the incubation. These results indicate that fresh human milk contains immunoreactive somatomedin-C, of which a significant proportion is not protein bound at neutral pH. Somatomedin-C may account for some of the mitogenic activity of milk reported by others. The role of this activity in neonatal growth is unknown.