METHODS FOR PRODUCING SUBSTANTIALLY HOMOGENEOUS HYBRID OR COMPLEX N-GLYCANS IN METHYLOTROPHIC YEASTS
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Issued Date -
Feb 2, 2012
app pub date -
Sep 23, 2011
filing date -
Oct 16, 2009
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Abstract
The present invention provides methods for effectively and efficiently converting methylotrophic yeast's heterogeneous high mannose-type N-glycosylation to mammalian-type N-glycosylation by disruption of an endogenous glycosyltransferase gene (OCH1) and step-wise introduction of heterologous glycosidase and glycosyltransferase activities. Each engineering step includes a number of stages: transformation with an appropriate vector, cultivation of a number of transformants, performance of sugar analysis and heterologous protein expression analysis, and selection of a desirable clone. The selected clone is then subjected to the next engineering step.
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