Open AccessRecombinant glycodelin carrying the same type of glycan structures as contraceptive glycodelin‐A can be produced in human kidney 293 cellsbut not in Chinese hamster ovary cells
Author(s) -
Van den Nieuwenhof Ingrid M.,
Koistinen Hannu,
Easton Richard L.,
Koistinen Riitta,
Kämäräinen Meerit,
Morris Howard R.,
van Die Irma,
Seppälä Markku,
Dell Anne,
Van den Eijnden Dirk H.
Publication year - 2000
Publication title -
european journal of biochemistry
Language(s) - English
Resource type - Journals
eISSN - 1432-1033
pISSN - 0014-2956
DOI - 10.1046/j.1432-1327.2000.01528.x
Subject(s) - chinese hamster ovary cell , recombinant dna , glycan , glycosylation , cell culture , biology , baby hamster kidney cell , hek 293 cells , kidney , cricetulus , microbiology and biotechnology , biochemistry , chemistry , cell , glycoprotein , endocrinology , genetics , gene
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 β1→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.