Recombinant expression and purification of the antimicrobial peptide magainin‐2

Ramos, Reinaldo Rodrigues; Moreira, Susana Margarida Gomes; Rodrigues, Ana Cristina; Gama, Miguel; Domingues, Lucı́lia

doi:10.1002/btpr.1650

Cited by 40 publications

(28 citation statements)

References 38 publications

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“…It is possible to make fully synthetic peptides by chemical synthesis [51] or by using recombinant expression systems [52,53]. These artificial sources of AMPs are useful for modification of existing AMPs and for designing new synthetic AMPs.…”

Section: Structure and Major Activities Of Ampsmentioning

confidence: 99%

Antimicrobial Peptides

2013

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The rapid increase in drug-resistant infections has presented a serious challenge to antimicrobial therapies. The failure of the most potent antibiotics to kill “superbugs” emphasizes the urgent need to develop other control agents. Here we review the history and new development of antimicrobial peptides (AMPs), a growing class of natural and synthetic peptides with a wide spectrum of targets including viruses, bacteria, fungi, and parasites. We summarize the major types of AMPs, their modes of action, and the common mechanisms of AMP resistance. In addition, we discuss the principles for designing effective AMPs and the potential of using AMPs to control biofilms (multicellular structures of bacteria embedded in extracellular matrixes) and persister cells (dormant phenotypic variants of bacterial cells that are highly tolerant to antibiotics).

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Section: Structure and Major Activities Of Ampsmentioning

confidence: 99%

Antimicrobial Peptides

2013

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“…Fusion partners can also be of use when producing toxic proteins. An example is the production of antimicrobial peptides (AMPs) by E. coli using cellulose binding modules as fusion partner (Guerreiro et al, 2008; Ramos et al, 2010, 2013). The use of carbohydrate-binding modules (CBMs) as fusion partner has also been applied for targeting peptides and/or functionalizing specific supports/biomaterials for biomedical applications (Moreira et al, 2008; Andrade et al, 2010a,b; Pértile et al, 2012).…”

Section: Soluble Protein Production In Escherichia Colimentioning

confidence: 99%

Fusion tags for protein solubility, purification and immunogenicity in Escherichia coli: the novel Fh8 system

et al. 2014

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Proteins are now widely produced in diverse microbial cell factories. The Escherichia coli is still the dominant host for recombinant protein production but, as a bacterial cell, it also has its issues: the aggregation of foreign proteins into insoluble inclusion bodies is perhaps the main limiting factor of the E. coli expression system. Conversely, E. coli benefits of cost, ease of use and scale make it essential to design new approaches directed for improved recombinant protein production in this host cell. With the aid of genetic and protein engineering novel tailored-made strategies can be designed to suit user or process requirements. Gene fusion technology has been widely used for the improvement of soluble protein production and/or purification in E. coli, and for increasing peptide’s immunogenicity as well. New fusion partners are constantly emerging and complementing the traditional solutions, as for instance, the Fh8 fusion tag that has been recently studied and ranked among the best solubility enhancer partners. In this review, we provide an overview of current strategies to improve recombinant protein production in E. coli, including the key factors for successful protein production, highlighting soluble protein production, and a comprehensive summary of the latest available and traditionally used gene fusion technologies. A special emphasis is given to the recently discovered Fh8 fusion system that can be used for soluble protein production, purification, and immunogenicity in E. coli. The number of existing fusion tags will probably increase in the next few years, and efforts should be taken to better understand how fusion tags act in E. coli. This knowledge will undoubtedly drive the development of new tailored-made tools for protein production in this bacterial system.

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“…[217,218] The most used CBMs are those that have affinity for cellulose, which can be used as affinity tags for purification and immobilization of peptides and proteins. [219–221] Ofir et al prepared a series of scFv antibody fragments coupled with cellulose-binding domains and used them to construct a CBM-based microarray for HIV serodiagnosis. [222,223]…”

Section: Antibody Immobilization Chemistriesmentioning

confidence: 99%

Site-selective orientated immobilization of antibodies and conjugates for immunodiagnostics development

Shen

Rusling

Dixit

2017

Methods

157

109

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Immobilized antibody systems are the key to develop efficient diagnostics and separations tools. In the last decade, developments in the field of biomolecular engineering and crosslinker chemistry have greatly influenced the development of this field. With all these new approaches at our disposal, several new immobilization methods have been created to address the main challenges associated with immobilized antibodies. Few of these challenges that we have discussed in this review are mainly associated to the site-specific immobilization, appropriate orientation, and activity retention. We have discussed the effect of antibody immobilization approaches on the parameters on the performance of an immunoassay.

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Recombinant expression and purification of the antimicrobial peptide magainin‐2

Cited by 40 publications

References 38 publications

Antimicrobial Peptides

Antimicrobial Peptides

Fusion tags for protein solubility, purification and immunogenicity in Escherichia coli: the novel Fh8 system

Site-selective orientated immobilization of antibodies and conjugates for immunodiagnostics development

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