Preparative method of functional oligasaccharides from bovine colostrum

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Colostrum was taken from a Holstein-Friesian cow 6h after parturition. Isolation of milk sugars from defatted colostrum by dialysis was fermormed by the method of Schneir and Rafelson. The complete removal of peptides from the dialyzate was achieved by ion-exchange chromatography with Sp-Sephadex C-50 by our earlier method. The sugars remaining after removal of lactose by crystallization were fractionated by gel filtration chromatography(Bio-gel P-2) with distilled water as eluent. On the otherhand, the colostrum glycopeptide was treated with NaOH of NaB <H_4> for 36h at 50 C in the dark under <N_2> , followed by removal of low-molecular peptides and salts using Dowex 50W-X4 resin. The resulting oligosacchariditols were finally purified by gel filtration chromatography(Biogel P-2) with distilled water as eluent. The purified oligosaccharditols were fractionated by ion-exchange chromatography using the method of Farrar et al in a linear concentration gradient of 0.05-1.0M of pyridine-acetate buffer.