The goat mammary glandular epithelial (GMGE) cell line promotes polyfucosylation and N,N '-diacetyllactosediaminylation of N-glycans linked to recombinant human erythropoietin
We have established a continuous, non-transformed cell line from primary cultures from Capra hircus mammary gland. Low-density cultures showed a homogeneous epithelial morphology without detectable fibroblastic or myoepithelial cells. The culture was responsive to contact inhibition of proliferation and its doubling tirne was dependent on the presence of insulin and epidermal growth factor (EGF). GMGE cells secrete caseins regardless of the presence or absence of lactogenic hormones in the culture media. Investigation of the total N-glycan pool of human erythropoietin (rhEPO) expressed in GNIGE cells by monosaccharide analysis, HPLC profiling, and mass spectrometry, indicated significant differences with respect to the same protein expressed in Chinese hamster ovary (CHO) cells. N-Glycans of rhEPO-GMGE are core-fitcosylated, but fucosylation of outer arms was also found. Our results also revealed the presence of low levels of sialylation (> 95%Neu5Ac), N,N'diacetyllactosediamine units, and possibly Gal-Gal non-reducing terminal elements. (c) 2007 Elsevier Inc. All rights reserved.
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|ARCHIVES OF BIOCHEMISTRY AND BIOPHYSICS
|Elsevier Science Inc.
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