Unique structure of glycopeptide from alpha-fetoprotein produced in human hepatoma cell line, as determined by 1H-nuclear magnetic resonance spectroscopy.
Determination of alpha-fetoprotein is used in diagnosis of tumors and neural tube defects. A good reliable source of alpha-fetoprotein would be an obvious advantage to the preparation of diagnostic reagents and their standardization. We have recently developed a method for the production of alpha-fetoprotein from a human hepatoma cell line. This method, which is suitable for scaling up, allowed us to produce 40 g of alpha-fetoprotein from culture supernatant liquid through a simple purification procedure. We have previously shown this protein to be identical to alpha-fetoprotein produced from other sources. However, because the presence of different glycoforms has been reported in alpha-fetoprotein preparations, both from human sources and from other species, it was important to establish the type and extent of glycosylation of alpha-fetoprotein prepared by our method. By using 1H-NMR spectroscopy we were able to establish that our product contains a single N-linked biantennary, fully sialylated complex-type oligosaccharide, typical of human hepatomas.
Centro Ricerche Sclavo, Siena, Italy.
B
B
Terrana
M F
MF
Tecce
R
R
Manetti
C
C
Ceccarini
D
D
Lamba
A L
AL
Segre
D006528:Carcinoma, Hepatocellular / Q000032:analysis*; D002460:Cell Line; D006020:Glycopeptides / Q000032:analysis*; D006801:Humans; D008113:Liver Neoplasms / Q000032:analysis*; D009682:Magnetic Resonance Spectroscopy; D011522:Protons; D000509:alpha-Fetoproteins / Q000032:analysis*
D006020:Glycopeptides; D011522:Protons; D000509:alpha-Fetoproteins