Recovery of intracellular recombinant proteins from the yeast Pichia pastoris by cell permeabilization

Shepard, Scot R.; Stone, Charmaine; Cook, Susan M.; Bouvier, Anne; Boyd, Gregory A.; Weatherly, Gresham T.; Lydiard, Debra D.; Schrimsher, Jeffrey L.

doi:10.1016/s0168-1656(02)00182-7

Cited by 18 publications

(8 citation statements)

References 12 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…Therefore, the clear extract contains by far less host cell contaminants compared to cleared cell homogenate. As an alternative to cell homogenization, extraction of intracellular proteins from whole cells has been reported [127]. For clarification and protein purification purposes, a wide variety of filters for protein processing is meanwhile available.…”

Section: Primary Separationmentioning

confidence: 99%

Manufacturing of recombinant therapeutic proteins in microbial systems

Graumann

Premstaller

2006

Biotechnology Journal

165

View full text Add to dashboard Cite

Recombinant therapeutic proteins have gained enormous importance for clinical applications. The first recombinant products have been produced in E. coli more than 20 years ago. Although with the advent of antibody-based therapeutics mammalian expression systems have experienced a major boost, microbial expression systems continue to be widely used in industry. Their intrinsic advantages, such as rapid growth, high yields and ease of manipulation, make them the premier choice for expression of non-glycosylated peptides and proteins. Innovative product classes such as antibody fragments or alternative binding molecules will further expand the use of microbial systems. Even more, novel, engineered production hosts and integrated technology platforms hold enormous potential for future applications. This review summarizes current applications and trends for development, production and analytical characterization of recombinant therapeutic proteins in microbial systems.

show abstract

Section: Primary Separationmentioning

confidence: 99%

Manufacturing of recombinant therapeutic proteins in microbial systems

Graumann

Premstaller

2006

Biotechnology Journal

165

View full text Add to dashboard Cite

show abstract

“…There are several examples of utilisation of chemical permeabilisation for protein recovery from yeast (Naglak et al, 1990, Shepard et al, 2002and Garcia, 2013. Most often, the permeabilisation is achieved by treatment with solvents or detergents.…”

Section: Discussionmentioning

confidence: 99%

Electroinduced release of recombinant β-galactosidase from Saccharomyces cerevisiae

Ganeva

Stefanova

Angelova

et al. 2015

Journal of Biotechnology

View full text Add to dashboard Cite

“…The most common methods used for P. pastoris cell lysis employ glass beads, microfluidizers, and a French press (Invitrogen 2005) [3,4], which are the more suitable choices for large amount of cells. Alternatively, Shepard et al [75] reported in 2002 a method for P. pastoris intracellular protein recovery based on cell permeabilization using N,N-dimethyltetradecylamine and Triton X-100.…”

Section: Biosynthesis Strategies For Protein Production In Pichia Pasmentioning

confidence: 99%

Pichia pastoris: A Recombinant Microfactory for Antibodies and Human Membrane Proteins

Gonçalves

Pedro²,

Maia³

et al. 2013

J. Microbiol. Biotechnol.

View full text Add to dashboard Cite

During the last few decades, it has become evident that the compatibility of the yeast biochemical environment with the ability to process and translate the RNA transcript, along with its capacity to modify a translated protein, are relevant requirements for selecting this host cell for protein expression in several pharmaceutical and clinical applications. In particular, Pichia pastoris is used as an industrial host for recombinant protein and metabolite production, showing a powerful capacity to meet required biomolecular target production levels in high-throughput assays for functional genomics and drug screening. In addition, there is a great advantage to using P. pastoris for protein secretion, even at high molecular weights, since the recovery and purification steps are simplified owing to relatively low levels of endogenous proteins in the extracellular medium. Clearly, no single microexpression system can provide all of the desired properties for human protein production. Moreover, chemical and physical bioprocess parameters, including culture medium formulation, temperature, pH, agitation, aeration rates, induction, and feeding strategies, can highly influence product yield and quality. In order to benefit from the currently available wide range of biosynthesis strategies using P. pastoris, this mini review focuses on the developments and technological fermentation achievements, providing both a comparative and an overall integration analysis. The main aim is to highlight the relevance and versatility of the P. pastoris biosystem to the design of more cost-effective microfactories to meet the increasing demands for recombinant membrane proteins and clinical antibodies for several therapeutic applications.

show abstract

Recovery of intracellular recombinant proteins from the yeast Pichia pastoris by cell permeabilization

Cited by 18 publications

References 12 publications

Manufacturing of recombinant therapeutic proteins in microbial systems

Manufacturing of recombinant therapeutic proteins in microbial systems

Electroinduced release of recombinant β-galactosidase from Saccharomyces cerevisiae

Pichia pastoris: A Recombinant Microfactory for Antibodies and Human Membrane Proteins

Contact Info

Product

Resources

About