Journal ArticleDOI
An optimized fermentation process for high-level production of a single-chain Fv antibody fragment in Pichia pastoris.
Leonardo M. Damasceno,Itzcoatl A. Pla,Hyun-Joo Chang,Leonard Cohen,Gerd Ritter,Lloyd J. Old,Carl A. Batt +6 more
TLDR
The expression of a humanized single-chain variable domain fragment antibody (A33scFv) was optimized for Pichia pastoris with yields exceeding 4 g L(-1) and demonstrates that functional A33 scFv can be produced in sufficient quantities using P. pastoris.About:
This article is published in Protein Expression and Purification.The article was published on 2004-09-01. It has received 124 citations till now. The article focuses on the topics: Pichia pastoris & Pichia.read more
Citations
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Journal ArticleDOI
Survey of the year 2003 commercial optical biosensor literature
Rebecca L. Rich,David G. Myszka +1 more
TL;DR: In this overview, 13 papers that should be on everyone's ‘must read’ list for 2003 are spotlighted and examples of how to identify and interpret high‐quality biosensor data are provided.
Journal ArticleDOI
Protein secretion in Pichia pastoris and advances in protein production.
TL;DR: Endoplasmic reticulum protein folding, correct glycosylation, vesicular transport to the plasma membrane, gene dosage, secretion signal sequences, and secretome studies are important considerations for improved recombinant protein production.
Journal ArticleDOI
Single chain Fab (scFab) fragment
Michael Hust,Thomas Jostock,Thomas Jostock,Christian Menzel,Christian Menzel,Bernd Voedisch,Anja Mohr,Anja Mohr,Mariam Brenneis,Mariam Brenneis,Martina Inga Kirsch,Doris Meier,Stefan Dübel +12 more
TL;DR: It is demonstrated that the introduction of a polypeptide linker between the fragment difficult (Fd) and the light chain (LC), resulting in the formation of a single chain Fab fragment (scFab), can lead to improved production of functional molecules.
Journal ArticleDOI
Emerging Technologies for Low-Cost, Rapid Vaccine Manufacture.
TL;DR: The assessment indicated that technologies in the order of increasing feasibility and decreasing risk are the yeast platform, ADDomer platform, followed by RNA and GMMA platforms.
Journal ArticleDOI
Gold hybrid nanoparticles for targeted phototherapy and cancer imaging
TL;DR: Gold and iron oxide hybrid nanoparticles synthesized by the thermal decomposition technique are bio-functionalized with a single chain antibody, scFv, that binds to the A33 antigen present on colorectal cancer cells to suggest that this new class of bio-conjugated hybrid nanoparticle can potentially serve as an effective antigen-targeted photothermal therapeutic agent for cancer treatment as well as a probe for magnetic resonance-based imaging.
References
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Book
Basic methods in molecular biology
TL;DR: General methods bacterial strains and cloning vectors enzymes that modify DNA and RNA in vitro amplification of DNA using the polymerase chain reaction (PCR) and the thermostable Taq DNA polymerase, introduction DNA restriction fragment analysis and preparation.
Journal ArticleDOI
Recent advances in the expression of foreign genes in Pichia pastoris
TL;DR: The Pichia pastoris heterologous gene expression system has been utilized to produce attractive levels of a variety of intracellular and extracellular proteins of interest and improvements in understanding and application have improved its utility even further.
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Production of recombinant proteins in fermenter cultures of the yeast Pichia pastoris.
TL;DR: The Pichia pastoris expression system offers economy, ease of manipulation, the ability to perform complex post-translational modifications, and high expression levels.
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Clinical evidence of efficient tumor targeting based on single-chain Fv antibody selected from a combinatorial library.
Richard H. J. Begent,M. J. Verhaar,KA Chester,Joanne L. Casey,AJ Green,M.P. Napier,L.D Hope‐Stone,N. Cushen,P. A. Keep,CJ Johnson,Robert E. Hawkins,Andrew J.W. Hilson,Lisa Robson +12 more
TL;DR: The benefits of this system for cancer targeting based on single–chain Fv (scFv) antibodies selected from combinatorial libraries, produced in bacteria and purified by using an engineered tag are illustrated.
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Advances in the use of Pichia pastoris for high-level gene expression
TL;DR: The Pichia pastoris system has the potential for very high level production of foreign proteins and is now also available for optimizing secretion, such as the generation of clones with progressively increasing vector copy number, expression screening in microtitre plates, and minimizing proteolysis by a number of techniques.