Document Detail


Array-based analysis of secreted glycoproteins for rapid selection of a single cell producing a glycoprotein with desired glycosylation.
MedLine Citation:
PMID:  20550136     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
The therapeutic efficacy and in vivo biological function of a glycoprotein is significantly affected by its glycosylation profile. For the development of glycoproteins with therapeutic applications, selection of cell lines producing a glycoprotein with adequate glycoform is crucial. Here, we demonstrate an array-based analysis of secreted glycoproteins for rapid and efficient selection of a single cell producing a glycoprotein with desirable glycosylation. Our approach relies on microengraving and interrogation of glycoproteins produced by individual cells in a microwell array in terms of glycosylation profile as well as the produced amount. On the basis of statistical analysis of the interrogation, single cells which are predicted to produce a desired glycoprotein are selected, retrieved, and expanded. We applied the approach to human recombinant erythropoietin (rhEPO)-producing CHO cells and verified the selection of a single CHO cell that produces rhEPO with a high sialylation degree. Human erythropoietin (hEPO) bearing highly sialylated oligosaccharide was shown to display a much longer plasma half-life, resulting in high therapeutic efficacy. This method may find widespread use in the clonal selection for the production of other glycoproteins with specific glycosylation as well as analysis of the heterogeneity in cell populations in a high-throughput manner.
Authors:
Sunyoung Park; Wanjung Kim; Yongtae Kim; Young Dok Son; Sang-Chul Lee; Eunkyung Kim; Sung Ho Kim; Jung Hoe Kim; Hak-Sung Kim
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Analytical chemistry     Volume:  82     ISSN:  1520-6882     ISO Abbreviation:  Anal. Chem.     Publication Date:  2010 Jul 
Date Detail:
Created Date:  2010-06-30     Completed Date:  2010-09-29     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  0370536     Medline TA:  Anal Chem     Country:  United States    
Other Details:
Languages:  eng     Pagination:  5830-7     Citation Subset:  IM    
Affiliation:
Departments of Biological Sciences, Korea Advanced Institute of Science and Technology, 373-1, Guseong-dong, Yuseong-gu, Daejeon 305-701, Korea.
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MeSH Terms
Descriptor/Qualifier:
Animals
CHO Cells
Cricetinae
Cricetulus
Erythropoietin, Recombinant / chemistry,  genetics,  metabolism
Fluorometry
Glycoproteins / analysis*,  chemistry
Glycosylation
Half-Life
High-Throughput Screening Assays
Humans
Protein Array Analysis / methods*
Chemical
Reg. No./Substance:
0/Erythropoietin, Recombinant; 0/Glycoproteins

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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