Recombinant glycodelin carrying the same type of glycan structures as contraceptive glycodelin-A can be produced in human kidney 293 cells but not in chinese hamster ovary cells.
We have produced human recombinant glycodelin in human kidney 293 cells and in Chinese hamster ovary (CHO) cells. Structural analyses by lectin immunoassays and fast atom bombardment mass spectrometry showed that recombinant human glycodelin produced in CHO cells contains only typical CHO-type glycans and is devoid of any of the N, N'-diacetyllactosediamine (lacdiNAc)-based chains previously identified in glycodelin-A (GdA). By contrast, human kidney 293 cells produced recombinant glycodelin with the same type of carbohydrate structures as GdA. The presence of a beta1-->4-N-acetylgalactosaminyltransferase functioning in the synthesis of lacdiNAc-based glycans in human kidney 293 cells is concluded to be the cause of the occurrence of lacdiNAc-based glycans on glycodelin produced in these cells. Furthermore, human kidney 293 cells were found to be particularly suited for the production of recombinant glycodelin when they were cultured in high glucose media. Lowering the glucose concentration and the addition of glucosamine resulted in higher relative amounts of oligomannosidic-type glycans and complex glycans with truncated antennae. Human glycodelin is an attractive candidate for the development of a contraceptive agent, and this study gives valuable information for selecting the proper expression system and cell culture conditions for the production of a correctly glycosylated recombinant form.
Van den Nieuwenhof
I M
IM
Department of Medical Chemistry, Faculty of Medicine, Vrije Universiteit, Amsterdam, The Netherlands. IM.van_den_Nieuwenhof.medchem@med.vu.nl
Koistinen
H
H
Easton
R L
RL
Koistinen
R
R
Kämäräinen
M
M
Morris
H R
HR
Van Die
I
I
Seppälä
M
M
Dell
A
A
Van den Eijnden
D H
DH
D000818:Animals; D016466:CHO Cells; D002240:Carbohydrate Sequence; D018929:Cell Culture Techniques / Q000379:methods; D002460:Cell Line; D003270:Contraceptive Agents / Q000737:chemistry / Q000378:metabolism*; D006224:Cricetinae; D018076:DNA, Complementary / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D005944:Glucosamine / Q000494:pharmacology; D005947:Glucose / Q000494:pharmacology; D000071816:Glycodelin; D006023:Glycoproteins / Q000096:biosynthesis* / Q000737:chemistry; D006031:Glycosylation; D006801:Humans; D007118:Immunoassay; D015151:Immunoblotting; D037102:Lectins / Q000737:chemistry; D008969:Molecular Sequence Data; D009005:Monosaccharides / Q000737:chemistry; D011257:Pregnancy Proteins / Q000096:biosynthesis* / Q000737:chemistry; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000737:chemistry; D016339:Spectrometry, Mass, Fast Atom Bombardment; D014162:Transfection
D003270:Contraceptive Agents; D018076:DNA, Complementary; D000071816:Glycodelin; D006023:Glycoproteins; D037102:Lectins; D009005:Monosaccharides; C045246:PAEP protein, human; D011257:Pregnancy Proteins; D011994:Recombinant Proteins; D005947:Glucose; D005944:Glucosamine