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Optimisation and evaluation of a high-throughput mammalian protein expression system.
- Source :
-
Protein expression and purification [Protein Expr Purif] 2005 Jul; Vol. 42 (1), pp. 111-21. Date of Electronic Publication: 2005 Mar 31. - Publication Year :
- 2005
-
Abstract
- Transient transfection of mammalian cells with episomal vectors is a very useful method for producing high levels of recombinant proteins. Transient systems remove the need for the laborious and time-consuming process of creating stable cell lines. Here, we describe the optimisation and evaluation of a high-throughput transient expression system in HEK293-EBNA cells. The process was developed for the expression of 10 constructs simultaneously in deep-well plates and subsequent purification using 96-well plate affinity chromatography. This enabled multiple combinations of different constructs, vectors, and expression conditions to be studied in parallel.
- Subjects :
- Animals
Carboxypeptidase B2 biosynthesis
Carboxypeptidase B2 genetics
Carboxypeptidase B2 isolation & purification
Cell Culture Techniques methods
Cell Line
Cell Proliferation drug effects
Epstein-Barr Virus Nuclear Antigens genetics
Genetic Vectors genetics
Glutathione Transferase biosynthesis
Glutathione Transferase genetics
Glutathione Transferase isolation & purification
Green Fluorescent Proteins biosynthesis
Green Fluorescent Proteins genetics
Green Fluorescent Proteins isolation & purification
Histidine genetics
Humans
Lymphocyte Specific Protein Tyrosine Kinase p56(lck) biosynthesis
Lymphocyte Specific Protein Tyrosine Kinase p56(lck) genetics
Lymphocyte Specific Protein Tyrosine Kinase p56(lck) isolation & purification
Poloxamer pharmacology
Protein Serine-Threonine Kinases biosynthesis
Protein Serine-Threonine Kinases genetics
Protein Serine-Threonine Kinases isolation & purification
Protein-Tyrosine Kinases biosynthesis
Protein-Tyrosine Kinases genetics
Protein-Tyrosine Kinases isolation & purification
Recombinant Proteins isolation & purification
Reproducibility of Results
Transfection methods
Gene Expression genetics
Recombinant Proteins biosynthesis
Subjects
Details
- Language :
- English
- ISSN :
- 1046-5928
- Volume :
- 42
- Issue :
- 1
- Database :
- MEDLINE
- Journal :
- Protein expression and purification
- Publication Type :
- Academic Journal
- Accession number :
- 15939296
- Full Text :
- https://doi.org/10.1016/j.pep.2005.03.012