ArticlePloS one2018
Titer estimation for quality control (TEQC) method: A practical approach for optimal production of protein complexes using the baculovirus expression vector system.
Article in PloS one, 2018. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 9 papers.
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Who cites it
9 citing papers in PubMed.
- Equi-MOI ratio for rapid baculovirus-mediated multiprotein co-expression in insect cells integrating selenomethionine for structural studies.FEBS open bio · 2025Article
- 3D variability analysis reveals a hidden conformational change controlling ammonia transport in human asparagine synthetase.Nature communications · 2024Article
- 3D Variability Analysis Reveals a Hidden Conformational Change Controlling Ammonia Transport in Human Asparagine Synthetase.bioRxiv : the preprint server for biology · 2024Article
- Isha is a su(Hw) mRNA-binding protein required for gypsy insulator function.G3 (Bethesda, Md.) · 2022Article
- CAMSAP2 organizes a γ-tubulin-independent microtubule nucleation centre through phase separation.eLife · 2022Article
- Structure and RNA template requirements ofProceedings of the National Academy of Sciences of the United States of America · 2021Article
- A practical method for efficient and optimal production of Seleno-methionine-labeled recombinant protein complexes in the insect cells.Protein science : a publication of the Protein Society · 2019Article
- High-resolution crystal structure of human asparagine synthetase enables analysis of inhibitor binding and selectivity.Communications biology · 2019Article
- A protein-protein interaction underlies the molecular basis for substrate recognition by an adenosine-to-inosine RNA-editing enzyme.Nucleic acids research · 2018Article
Corrections and comments
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Authors and funding
5 authors.
Funding
Abstract
The baculovirus expression vector system (BEVS) is becoming the method of choice for expression of many eukaryotic proteins and protein complexes for biochemical, structural and pharmaceutical studies. Significant technological advancement has made generation of recombinant baculoviruses easy, efficient and user-friendly. However, there is a tremendous variability in the amount of proteins made using the BEVS, including different batches of virus made to express the same proteins. Yet, what influences the overall production of proteins or protein complexes remains largely unclear. Many downstream applications, particularly protein structure determination, require purification of large quantities of proteins in a repetitive manner, calling for a reliable experimental set-up to obtain proteins or protein complexes of interest consistently. During our investigation of optimizing the expression of the Mediator Head module, we discovered that the 'initial infectivity' was an excellent indicator of overall production of protein complexes. Further, we show that this initial infectivity can be mathematically described as a function of multiplicity of infection (MOI), correlating recombinant protein yield and virus titer. All these findings led us to develop the Titer Estimation for Quality Control (TEQC) method, which enables researchers to estimate initial infectivity, titer/MOI values in a simple and affordable way, and to use these values to quantitatively optimize protein expressions utilizing BEVS in a highly reproducible fashion.
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