Journal Article10.1016/S1389-1723(03)80019-1
Stabilization of affinity-tagged recombinant protein during/after its production in a cell-free system using wheat-germ extract.
Yasuaki Kawarasaki,Yasuhiro Yamada,Maki Ichimori,Tomoya Shinbata,Katsunori Kohda,Hideo Nakano,Tsuneo Yamane +6 more
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TL;DR: It is found that the affinity tag fused to the carboxyl (C-) terminal of a single-chain Fv (scFv) antibody was proteolytically degraded in a wheat germ cell-free protein synthesis system, suggesting that wheat endogenous carboxypeptidase(s) play a primary role in the C-terminal tag-specific degradation.
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About: This article is published in Journal of Bioscience and Bioengineering. The article was published on 01 Jan 2003. The article focuses on the topics: Myc-tag & FLAG-tag.
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Citations
Construction and analysis of variants of a polyvalent Lyme disease vaccine: approaches for improving the immune response to chimeric vaccinogens.
TL;DR: Analyses of murine antibody titers and isotype profiles induced by constructs revealed that while the C-terminal tags did not enhance antibody titer, specific epitope reorganization and reiteration did, which provides important information that can be exploited in the development of chimeric vaccinogens in general.
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Patent
Polyvalent chimeric ospc vaccinogen and diagnostic antigen
Richard T. Marconi,Christopher G. Earnhart +1 more
- 29 Nov 2006
TL;DR: A chimeric polyvalent recombinant protein for use as a vaccine and diagnostic for Lyme disease was provided in this paper, which comprises epitopes of the loop 5 region and/or the alpha helix 5 region of outer surface protein C (OspC) types.
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Properties of recombinant Strep -tagged and untagged hyperthermophilic D-arabitol dehydrogenase from Thermotoga maritima
TL;DR: The first hyperthermophilic d-arabitol dehydrogenase from Thermotoga maritima was heterologously purified from Escherichia coli and had similar kinetic parameters compared to the tagged enzyme, demonstrating that the Strep-tag was not deleterious to protein function but decreased protein stability.
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Improvements in the cell-free production of functional antibodies using cell extract from protease-deficient Escherichia coli mutant
TL;DR: The results suggest that the DegP- and OmpT-deleted mutant is a useful source of S30 extract for the production or screening of antibodies using the cell-free expression system.
A plant derived multifunctional tool for nanobiotechnology based on Tomato bushy stunt virus
TL;DR: The ability of this system to remarkably sustain genetic modifications and in vitro chemical derivatizations of its outer surface, which resulted in the successful display of large chimeric peptides fusions and small chemical molecules, respectively is demonstrated.
References
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Molecular cloning : a laboratory manual
Joseph Sambrook,David W. Russell +1 more
- 01 Jan 2001
TL;DR: The content has been entirely recast to include nucleic-acid based methods selected as the most widely used and valuable in molecular and cellular biology laboratories.
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Single Step Generation of Protein Arrays From DNA by Cell-Free Expression and in Situ Immobilisation (PISA Method)
Mingyue He,Michael J. Taussig +1 more
TL;DR: It is shown that human single-chain antibody fragments and an enzyme (luciferase) can be functionally arrayed by the PISA method, and this method can overcome problems of insolubility or degradation associated with bacterial expression of recombinant proteins.
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Recent advances in producing and selecting functional proteins by using cell-free translation
TL;DR: Progress has been made to efficiently produce proteins in batch or continuous cell-free translation systems and to elucidate the molecular causes of low yield and find possible solutions for this problem.
223
A gibberellin responsive wheat gene has homology to yeast carboxypeptidase Y.
TL;DR: It is shown here that for one of those cDNA clones from mRNAs which are produced in increased amounts when aleurone layers of wheat are treated with gibberellic acid, the change in level of mRNA in aleur one parallels the changeIn level of alpha-amylase mRNA.
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High-throughput, Cloning-independent Protein Library Construction by Combining Single-molecule DNA Amplification with in Vitro Expression
Suang Rungpragayphan,Yasuaki Kawarasaki,Takao Imaeda,Katsunori Kohda,Hideo Nakano,Tsuneo Yamane +5 more
TL;DR: A library of anti-human serum albumin single-chain antibodies originating from a monoclonal anti-HSA-scFv which was constructed and screened in order to demonstrate its real practicability and was better than the classical in vivo expression system.
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