Total casein subunits as well as whey proteins were quantitated in human milk samples during lactation. Two independent methods were used: precipitation at pH 4.3 in the presence of Ca2+ followed by Kjeldahl analysis and polyacrylamide gradient gel electrophoresis (PAGGE) followed by densitometric scanning. Both methods yielded similar results: casein synthesis is low or absent in early lactation, then increases rapidly and subsequently decreases. The concentration of whey proteins decreases from early lactation and continues to fall. These changes result in a whey protein/casein ratio of about 90:10 in early lactation, 60:40 in mature milk and 50:50 in late lactation. These observations indicate that the synthesis and/or secretion of caseins and whey proteins is regulated by different mechanisms. In addition, the relative proportion of the different beta- and kappa-casein subunits was found to vary throughout lactation.