Production of Recombinant Proteins for Therapy, Diagnostics, and Industrial Research by in Vitro Folding

Lange, Christian; Rudolph, Rainer

doi:10.1002/9783527619498.ch71

Cited by 11 publications

(4 citation statements)

References 161 publications

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“…Because of this, refolding of larger multi-domain proteins in vitro is inefficient in many cases [8]. However, in particular by employing cosolvent-assisted folding (sugars, osmolytes), artificial chaperone systems (detergent, cyclodextrin), and in vitro chaperonin systems (e.g., GroEL/GroES) increased folding efficiencies have been achieved with numerous multi-domain proteins [9,10]. Even more efficient folding was observed for de novo synthesized multi-domain proteins where individual domains are synthesized sequentially and, apparently, also fold sequentially [8].…”

Section: Introductionmentioning

confidence: 99%

The perspectives of studying multi-domain protein folding

Fitter

2009

Cell. Mol. Life Sci.

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Most of fundamental studies on protein folding have been performed with small globular proteins consisting of a single domain. In vitro many of these proteins are well characterized by a reversible two-state folding scheme. However, the majority of proteins in the cell belong to the class of larger multi-domain proteins that often unfold irreversibly under in vitro conditions. This makes folding studies difficult or even impossible. In spite of these problems for many multi-domain proteins, folding has been investigated by classical refolding. Co-translational folding of nascent polypeptide chains when synthesized by ribosomes has also been studied. Single molecule techniques represent a promising approach for future studies on the folding of multi-domain proteins, and tremendous advances have been made in these techniques in recent years. In particular, fluorescence-based methods can contribute significantly to an understanding of the fundamental principles of multi-domain protein folding.

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Section: Introductionmentioning

confidence: 99%

The perspectives of studying multi-domain protein folding

Fitter

2009

Cell. Mol. Life Sci.

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“…For quite some time it has been known that additives, typically low‐molecular‐weight organic compounds, may significantly enhance the yield of the refolding process. In many cases, inclusion body protein can only be successfully refolded by making use of this effect (for reviews, see, e.g., De Bernardez Clark et al 1999; Tsumoto et al 2003; Fahnert et al 2004; Lange and Rudolph 2005).…”

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confidence: 99%

Ionic liquids as refolding additives: N′‐alkyl and N′‐(ω‐hydroxyalkyl) N‐methylimidazolium chlorides

2005

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The purpose of this work was to investigate the influence of a series of N¢-alkyl and N¢-(v-hydroxyalkyl)-N-methylimidazolium chlorides on the renaturation of two model proteins, namely hen egg white lysozyme and the single-chain antibody fragment ScFvOx. All tested ionic liquids acted as refolding enhancers, with varying efficacies and efficiencies. The results of the refolding screening could be interpreted by taking into account the effect of the studied ionic liquids on protein aggregation, together with the systematic variations of their influence on the stability of native proteins in solution. More hydrophobic imidazolium cations carrying longer alkyl chains were increasingly destabilizing, while terminal hydroxylation of the alkyl chain made the salts more compatible with protein stability. The studied ionic liquids can be classified as preferentially bound, slightly to moderately chaotropic cosolvents for proteins.

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“…Refolding conditions have to be carefully chosen in order to obtain satisfactory amounts of active protein. For quite some time it has been known that the addition of certain small organic molecules may significantly enhance the yield of the refolding process, and in many cases the denaturation and refolding of inclusion body protein only becomes practical by making use of this effect (for recent reviews on this topic see, e.g., De Bernardez Clark et al 1999; Tsumoto et al 2003; Fahnert et al 2004; Lange and Rudolph 2005).…”

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l‐Arginine increases the solubility of unfolded species of hen egg white lysozyme

et al. 2005

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L-Arginine (L-Arg) has been widely used as an enhancer of protein renaturation. The mechanism behind its action is still not fully understood. Using hen egg white lysozyme as a model protein, we present data that clearly demonstrate the suppression of the aggregation of denatured protein by L-Arg. By chemical modification of free cysteines, a series of unfolded lysozyme species were obtained that served as models for unfolded and intermediate states during the process of oxidative refolding. An increased equilibrium solubility of unfolded species and intermediates in the presence of L-Arg seems to be its major mechanism of action.

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Production of Recombinant Proteins for Therapy, Diagnostics, and Industrial Research by in Vitro Folding

Cited by 11 publications

References 161 publications

The perspectives of studying multi-domain protein folding

The perspectives of studying multi-domain protein folding

Ionic liquids as refolding additives: N′‐alkyl and N′‐(ω‐hydroxyalkyl) N‐methylimidazolium chlorides

l‐Arginine increases the solubility of unfolded species of hen egg white lysozyme

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