DC Field | Value | Language |
---|---|---|
dc.contributor.author | Kim, SH | ko |
dc.contributor.author | Lee, Gyun-Min | ko |
dc.date.accessioned | 2013-03-06T23:18:01Z | - |
dc.date.available | 2013-03-06T23:18:01Z | - |
dc.date.created | 2012-02-06 | - |
dc.date.created | 2012-02-06 | - |
dc.date.issued | 2007-02 | - |
dc.identifier.citation | APPLIED MICROBIOLOGY AND BIOTECHNOLOGY, v.74, pp.152 - 159 | - |
dc.identifier.issn | 0175-7598 | - |
dc.identifier.uri | http://hdl.handle.net/10203/88770 | - |
dc.description.abstract | Lactate, one of the major waste products in mammalian cell culture, can inhibit cell growth and affect cellular metabolism at high concentrations. To reduce lactate formation, lactate dehydrogenase-A (LDH-A), an enzyme catalyzing the conversion of glucose-derived pyruvate to lactate, was down-regulated by an expression vector of small interfering RNAs (siRNA) in recombinant Chinese hamster ovary (rCHO) cells producing human thrombopoietin (hTPO). Three clones expressing low levels of LDH-A, determined by reverse transcription-PCR and an enzyme activity test, were established in addition to a negative control cell line. LDH-A activities in the three clones were decreased by 75-89%, compared with that of the control CHO cell line, demonstrating that the effect of siRNA is more significant than that of other traditional methods such as homologous recombination (30%) and antisense mRNA (29%). The specific glucose consumption rates of the three clones were reduced to 54-87% when compared to the control cell line. Similarly, the specific lactate production rates were reduced to 45-79% of the control cell line level. In addition, reduction of LDH-A did not impair either cell proliferation or hTPO productivity. Taken together, these results show that the lactate formation rate in rCHO cell culture can be efficiently reduced through the down-regulation of LDH via siRNA. | - |
dc.language | English | - |
dc.publisher | SPRINGER | - |
dc.subject | MONOCLONAL-ANTIBODY PRODUCTION | - |
dc.subject | SHORT INTERFERING RNAS | - |
dc.subject | PYRUVATE-CARBOXYLASE | - |
dc.subject | CULTURE MEDIA | - |
dc.subject | EXPRESSION | - |
dc.subject | METABOLISM | - |
dc.subject | REDUCTION | - |
dc.subject | GLUTAMINE | - |
dc.subject | DECOMPOSITION | - |
dc.subject | LINES | - |
dc.title | Down-regulation of lactate dehydrogenase-A by siRNAs for reduced lactic acid formation of Chinese hamster ovary cells producing thrombopoietin | - |
dc.type | Article | - |
dc.identifier.wosid | 000243802200019 | - |
dc.identifier.scopusid | 2-s2.0-33846643485 | - |
dc.type.rims | ART | - |
dc.citation.volume | 74 | - |
dc.citation.beginningpage | 152 | - |
dc.citation.endingpage | 159 | - |
dc.citation.publicationname | APPLIED MICROBIOLOGY AND BIOTECHNOLOGY | - |
dc.identifier.doi | 10.1007/s00253-006-0654-5 | - |
dc.contributor.localauthor | Lee, Gyun-Min | - |
dc.contributor.nonIdAuthor | Kim, SH | - |
dc.type.journalArticle | Article | - |
dc.subject.keywordAuthor | Chinese hamster ovary (CHO) cells | - |
dc.subject.keywordAuthor | lactate dehydrogenase-A (LDH-A) | - |
dc.subject.keywordAuthor | small interfering RNA (siRNA) | - |
dc.subject.keywordAuthor | specific lactate production rate | - |
dc.subject.keywordPlus | MONOCLONAL-ANTIBODY PRODUCTION | - |
dc.subject.keywordPlus | SHORT INTERFERING RNAS | - |
dc.subject.keywordPlus | PYRUVATE-CARBOXYLASE | - |
dc.subject.keywordPlus | CULTURE MEDIA | - |
dc.subject.keywordPlus | EXPRESSION | - |
dc.subject.keywordPlus | METABOLISM | - |
dc.subject.keywordPlus | REDUCTION | - |
dc.subject.keywordPlus | GLUTAMINE | - |
dc.subject.keywordPlus | DECOMPOSITION | - |
dc.subject.keywordPlus | LINES | - |
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