Tagging Recombinant Proteins to Enhance Solubility and Aid Purification

Walls, Dermot; Loughran, Sinéad T.

doi:10.1007/978-1-60761-913-0_9

Cited by 91 publications

(50 citation statements)

References 146 publications

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“…Multiple in-depth reviews of TAP tags and purification schemes are currently available [5,170,171]. Detailed elements of combinatorial cassettes used for dual-tagging and TAP strategies, as well as vectors available to the bacteria and yeast research communities, relevant methods, and protocols are provided in Supporting information, Tables S3 and S5.…”

Section: Tandem Affinity Purification and Dual-tagging Methodsmentioning

confidence: 99%

“…While contemporary reports have included several overviews of available affinity tags for protein detection or purification [1][2][3][4], solubility enhancement [5], tag removal [1], or applications [6,7] -in this comprehensive review, we provide an extensive summary of various tags and established purification strategies. We describe several design aspects for these tags that should be considered for recombinant fusion protein expression, including amino acid composition and size; N-or C-terminal fusions, used individually or in tandem; as well as optimized tags, techniques, and buffer conditions that lead to purified protein.…”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Recombinant protein expression and purification: A comprehensive review of affinity tags and microbial applications

Young

Britton

Robinson

2012

Biotechnology Journal

410

267

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Protein fusion tags are indispensible tools used to improve recombinant protein expression yields, enable protein purification, and accelerate the characterization of protein structure and function. Solubility-enhancing tags, genetically engineered epitopes, and recombinant endoproteases have resulted in a versatile array of combinatorial elements that facilitate protein detection and purification in microbial hosts. In this comprehensive review, we evaluate the most frequently used solubility-enhancing and affinity tags. Furthermore, we provide summaries of well-characterized purification strategies that have been used to increase product yields and have widespread application in many areas of biotechnology including drug discovery, therapeutics, and pharmacology. This review serves as an excellent literature reference for those working on protein fusion tags.

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Section: Tandem Affinity Purification and Dual-tagging Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Recombinant protein expression and purification: A comprehensive review of affinity tags and microbial applications

Young

Britton

Robinson

2012

Biotechnology Journal

410

267

View full text Add to dashboard Cite

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“…25,26 In addition to facilitating identification, affinity tags can also confer enhanced protein stability, increased expression, and facilitate native folding. 27,28 Disadvantages of affinity tags include their requirement of expensive resins as display supports, in addition to possible affinity tag removal. 29,30 N-terminal glutathione S-transferase (GST) is also commonly used as an affinity tag for the purification of recombinant antibodies.…”

Section: Affinity Chromatographymentioning

confidence: 99%

Technology advancements in antibody purification

Murphy¹,

Devine²,

O’Kennedy³

2016

ANTI

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Abstract:The separation of antibodies from complex mixtures can be achieved using chromatographic or non-chromatographic techniques. The purification of antibodies using chromatography involves the separation of antibodies or antibody-derived molecules present in complex mixtures by passing them through a solid phase (eg, silica resin or beads, monolithic columns, or cellulose membranes) and allowing the antibodies to bind or pass through depending on whether "bind-and-elute" or "flow-through" chromatographic methods are employed. Chromatographic methodologies can incorporate different separation techniques, such as affinity-tag binding, ion-exchange, size-exclusion chromatography, or immunoaffinity chromatography. However, in a concerted effort to become less reliant on chromatography-based separations owing to the high cost of large-scale production, non-chromatographic-based techniques such as precipitation, flocculation, crystallization, filtration, and aqueous two-phase partitioning are now becoming more popular. This review details current chromatographic and non-chromatographic methodo logies used for the purification of antibodies and expands on technological advancements and practical uses that have recently been reported.

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“…2 Co-expression with a fusion partner is one of the common strategies to improve the expression of some hard-to-express and hard-to-solubilize proteins in E. coli. 4,5 Many fusion partners such as E. coli maltose binding protein have the ability to increase the yield, enhance the solubility and even promote the proper folding of their fused proteins. As an agonist of innate immune system, Salmonella flagellin has been demonstrated to be a potent adjuvant for various antigens and widely tried in vaccine research and development.…”

Section: Introductionmentioning

confidence: 99%