Development of neoglycoproteins conjugated with natural oligosaccharides through carboxyl residues of proteins and its application to recombinant human interleukin 1.
In order to develop glycosylated cytokines, neoglycoproteins were synthesized utilizing naturally occurring oligosaccharides. High mannose type oligosaccharide-asparagine (Asn)s, containing Man8GN2-Asn, Man7GN2-Asn and Man6GN2-Asn, were obtained from quail ovalbumin and were coupled to bovine serum albumin (BSA) by carbodiimide-mediated coupling. Major reaction occurred between amino residues of oligosaccharide-Asns and carboxyl residues of BSA. Approximately one molecules of oligosaccharides-Asns were coupled to per molecule of BSA with 50% yield of glycosylation. Among the oligosaccharides, Man6GN2-Asn appeared to be conjugated predominantly. While this strategy was applied to recombinant human interleukin 1 alpha (IL-1 alpha), three molecules of oligosaccharide-Asns were introduced into per molecule of IL-1 with 10% yield of glycosylation.
Ishihara
H
H
Nagoya City University College of Nursing, Japan.
Chiba
T
T
Takeuchi
A
A
Hayashi
H
H
Takei
Y
Y
Takii
T
T
Onozaki
K
K
D000818:Animals; D002234:Carbodiimides; D002240:Carbohydrate Sequence; D003370:Coturnix; D006023:Glycoproteins / Q000138:chemical synthesis*; D006031:Glycosylation; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007375:Interleukin-1 / Q000378:metabolism*; D008969:Molecular Sequence Data; D009844:Oligosaccharides / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000378:metabolism*; D012710:Serum Albumin, Bovine / Q000378:metabolism
D002234:Carbodiimides; D006023:Glycoproteins; D007375:Interleukin-1; D009844:Oligosaccharides; D011993:Recombinant Fusion Proteins; C074139:1-ethyl-3-(3-dimethylaminoethyl)carbodiimide; D012710:Serum Albumin, Bovine