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rdf:type |
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lifeskim:mentions |
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pubmed:issue |
5
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pubmed:dateCreated |
1994-3-22
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pubmed:abstractText |
A variety of approaches to maximizing the production of recombinant human alpha 1-antitrypsin (AAT) in Chinese hamster ovary (CHO) cells have been investigated. The highly active and inducible human cytomegalovirus immediate early (IE) promoter/enhancer was used to drive transcription of a recombinant AAT gene in transiently transfected and stably transformed CHO cells. The AAT gene was modified to incorporate highly efficient 3'RNA processing signals from the herpes simplex virus type 2 IE gene 5, and optimal translational initiation signals were created by site-directed mutagenesis. The effect of flanking the recombinant gene with matrix attachment regions was investigated. Combinations of these modifications allowed secretion of up to 44 micrograms AAT/ml per day by cell lines growing in serum-rich medium. This could be increased to up to 100 micrograms AAT/ml per day upon chemical induction of expression by propionate, butyrate or hexamethylene bisacetamide. Cell lines adapted to grow in protein-free medium produced less AAT but still responded to chemical induction to secrete up to 14 micrograms/ml per day of readily purified AAT.
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pubmed:language |
eng
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pubmed:journal |
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pubmed:citationSubset |
B
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pubmed:chemical |
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pubmed:status |
MEDLINE
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pubmed:month |
Jan
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pubmed:issn |
0175-7598
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pubmed:author |
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pubmed:issnType |
Print
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pubmed:volume |
40
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pubmed:geneSymbol |
AAT
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pubmed:owner |
NLM
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pubmed:authorsComplete |
Y
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pubmed:pagination |
691-8
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pubmed:dateRevised |
2006-5-1
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pubmed:meshHeading |
pubmed-meshheading:7764427-Animals,
pubmed-meshheading:7764427-Base Sequence,
pubmed-meshheading:7764427-Biotechnology,
pubmed-meshheading:7764427-Butyric Acid,
pubmed-meshheading:7764427-Butyric Acids,
pubmed-meshheading:7764427-CHO Cells,
pubmed-meshheading:7764427-Cell Line, Transformed,
pubmed-meshheading:7764427-Cricetinae,
pubmed-meshheading:7764427-DNA,
pubmed-meshheading:7764427-Dimethyl Sulfoxide,
pubmed-meshheading:7764427-Gene Expression,
pubmed-meshheading:7764427-Genetic Vectors,
pubmed-meshheading:7764427-Humans,
pubmed-meshheading:7764427-Molecular Sequence Data,
pubmed-meshheading:7764427-Mutagenesis, Site-Directed,
pubmed-meshheading:7764427-Plasmids,
pubmed-meshheading:7764427-Propionic Acids,
pubmed-meshheading:7764427-Recombinant Proteins,
pubmed-meshheading:7764427-Transformation, Genetic,
pubmed-meshheading:7764427-alpha 1-Antitrypsin
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pubmed:year |
1994
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pubmed:articleTitle |
Approaches to maximizing stable expression of alpha 1-antitrypsin in transformed CHO cells.
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pubmed:affiliation |
National Science Laboratory, Scottish National Blood Transfusion Service, Edinburgh, UK.
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pubmed:publicationType |
Journal Article
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