STUDIES ON THE IDENTIFICATION OF A FOLATE COMPOUND of human serum.pdf

Journal of Clinical Investigation Vol. 41, No. 5, 1962 STUDIES ON THE IDENTIFICATION OF A FOLATE COMPOUND OF HUMAN SERUM* By VICTOR HERBERT, ALLAN R. LARRABEE t AND JOHN M. BUCHANAN (From the Thorndike Memorial Laboratory and Second and Fourth (Harvard) Medical Serv- ices, Boston City Hospital; Department of Medicine, Harvard Medical School, Boston; and the Division of Biochemistry, Department of Biology, Massachusetts Insti- tute of Technology, Cambridge, Mass.) (Submitted for publication May 10, 1961; accepted January 25, 1962) Human serum contains folate (pteroylgluta- removed by filtration through gauze. The material was mate) compounds that support the growth of then centrifuged for 5 minutes in an International clini- Lactobacillus casei but not Streptococcus faecalis cal centrifuge. The supernatant fluid was decanted and stirred with 4.5 g of Supercel for 30 minutes. After cen- or Leuconostoc citrovorum (Pediococcus cere- trifugation the supernatant solution was decanted into visiae) (1). It has been generally accepted that 25 ml of a suspension of calcium phosphate gel (0.31 g any compound having folic acid activity for L. per ml), stirred for 30 minutes, and centrifuged. The casei but not for S. faecalis must be a triglutamate supernatant fluid was treated with 2 g of Darco G 60 (or polyglutamate) (1-4). On the basis of this activated charcoal 1 per 100 ml of solution, mixed for 30 minutes, and centri