Opportunities and challenges of the tag-assisted protein purification techniques: Applications in the pharmaceutical industry

Gomari, Mohammad Mahmoudi; Saraygord‐Afshari, Neda; Farsimadan, Marziye; Rostami, Neda; Aghamiri, Seyed Mahmoud Reza; Farajollahi, Mohammad M

doi:10.1016/j.biotechadv.2020.107653

Cited by 40 publications

(21 citation statements)

References 231 publications

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“…A common approach to overcome inclusion body formations is the use of solubility tags. This strategy may not only increase soluble protein recovery but may also aid in several purification methods [ 48 , 49 ]. Taking into account that 80% of the investments spent in this area are directly related to the recovery and purification of proteins, new solutions that simplify these processes, such as efficient solubility tags, are essential in biotechnological progress [ 15 ].…”

Section: Discussionmentioning

confidence: 99%

Strategies for the Production of Soluble Interferon-Alpha Consensus and Potential Application in Arboviruses and SARS-CoV-2

Grabarz

Lopes

Barbosa

et al. 2021

Life

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Biopharmaceutical production is currently a multibillion-dollar industry with high growth perspectives. The research and development of biologically sourced pharmaceuticals are extremely important and a reality in our current healthcare system. Interferon alpha consensus (cIFN) is a non-natural synthetic antiviral molecule that comprises all the most prevalent amino acids of IFN-α into one consensus protein sequence. For clinical use, cIFN is produced in E. coli in the form of inclusion bodies. Here, we describe the use of two solubility tags (Fh8 and DsbC) to improve soluble cIFN production. Furthermore, we analyzed cIFN production in different culture media and temperatures in order to improve biopharmaceutical production. Our results demonstrate that Fh8-cIFN yield was improved when bacteria were cultivated in autoinduction culture medium at 30 °C. After hydrolysis, the recovery of soluble untagged cIFN was 58% from purified Fh8-cIFN molecule, fourfold higher when compared to cIFN recovered from the DsbC-cIFN, which achieved 14% recovery. The biological activity of cIFN was tested on in vitro model of antiviral effect against Zika, Mayaro, Chikungunya and SARS-CoV-2 virus infection in susceptible VERO cells. We show, for the first time, that cIFN has a potent activity against these viruses, being very low amounts of the molecule sufficient to inhibit virus multiplication. Thus, this molecule could be used in a clinical approach to treat Arboviruses and SARS-CoV-2.

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Section: Discussionmentioning

confidence: 99%

Strategies for the Production of Soluble Interferon-Alpha Consensus and Potential Application in Arboviruses and SARS-CoV-2

Grabarz

Lopes

Barbosa

et al. 2021

Life

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“…Different biomolecules are separated because of their different binding forces with metal ions. 40 IMAC with Nickel ions has been used to separate angiotensin-I-converting enzyme (ACE) inhibitory peptides in protein hydrolysates 41 and to enrich special proteins and peptides. 42 Moreover, modification of the IMAC medium with methoxypolyethylene glycol can avoid non-specific adsorption of the chromatographic medium to proteins present in hydrolysates.…”

Section: Isolation and Purification Of Oyster Peptidesmentioning

confidence: 99%

A comprehensive review of oyster peptides: Preparation, characterisation and bioactivities

Hao

Wang

Cao

et al. 2021

Reviews in Aquaculture

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In 1980, bioactive peptides were first described in a study on peptide biosynthesis in the neurointermediate lobe of Xenopus laevis. 1 Since then, bioactive peptides with beneficial physiological properties to living organisms have been widely studied. At present, bioactive peptides, including those obtained from marine and terrestrial environments, are a promising area for development for the international food industry. Marine bioactive peptides have broad potential use in the development of functional foods and as new therapeutics and also play an important role in improving culture efficiency.Oysters, the most cultured shellfish worldwide, are an important marine biological resource to humans. According to statistics reported by the Food and Agriculture Organization of the United Nations (2018; http://www.fao.org/fishe ry/stati stics/ en), global oyster production was around 6.1 million tons. The chemical composition of oysters is (based on dry flesh weight) as follows: protein (39.1%-53.1%); glycogen (21.6%-38.9%); and fat (7.8%-8.7%). 2 Methods commonly used for protein extraction include salting out, enzymatic hydrolysis and organic solvent-or/and pH-based protocols. The most abundant amino acids in oysters are taurine (Tau), glycine (Gly), alanine (Ala), aspartic acid (Asp), glutamic acid (Glu) and proline (Pro). Moreover, since eight essential amino acids account for

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“…Since inactivated whole-cell vaccines are usually expressed in a complex biological matrix with a wide range of impurities such as cell debris, host-cell-derived contaminants (e.g., proteins, DNA, and endotoxins) as well as viral non-structural proteins, causing difficulties in distinguishing infected from vaccinated animals, downstream processing must comply with strict purity requirements [22]. Additionally, relatively simple purification steps should enormously benefit the vaccine product development process as process scale-up, manufacturing cost, and consistent product quality can be better controlled [23]. A highly desirable purification technology with the potential to meet high-purity criteria in a single step is immobilized metal affinity chromatography (IMAC), which is based on the interaction between an immobilized metal ion (Cu 2+ , Zn 2+ , Ni 2+ , Fe 3+ , or Co 2+ ) and electron donor groups located on the surface of proteins.…”

Section: Introductionmentioning

confidence: 99%

Engineering His-Tagged Senecavirus A for One-Step Purification of Viral Antigens

et al. 2022

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Senecavirus A (SVA) is a picornavirus that causes vesicular disease in swine, and the inactivated vaccine is used to prevent and control SVA infection. To develop a new chromatography strategy for the purification and concentration of SVA vaccine antigens, we inserted a 6×His-tag at the VP1 C-terminal of the SVA/HLJ/CHA/2016 in an infectious clone to rescue a His-tagged SVA. The constructed and rescued recombinant virus, named as rSVA-His, exhibited similar growth kinetics to that of its parental virus. In addition, the expression of a 6×His-tag on the surface of SVA showed genetic stability in cell passages in vitro, which allowed one-step purification of SVA antigens by Ni2+ affinity columns. Furthermore, the immunogenicity of the inactivated rSVA-His was evaluated by inoculating rabbits and detecting neutralizing antibodies. The animals receiving two doses of the inactivated rSVA-His emulsified with oil adjuvant developed a high titer of neutralizing antibodies, indicating that SVA VP1 is tolerant to His-tag insertion without detriment to its antigenicity. In summary, the constructed 6×His-tagged SVA may offer a feasible approach to the affinity purification and concentration of antigens in the process of SVA inactivated vaccine production.

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Opportunities and challenges of the tag-assisted protein purification techniques: Applications in the pharmaceutical industry

Cited by 40 publications

References 231 publications

Strategies for the Production of Soluble Interferon-Alpha Consensus and Potential Application in Arboviruses and SARS-CoV-2

Strategies for the Production of Soluble Interferon-Alpha Consensus and Potential Application in Arboviruses and SARS-CoV-2

A comprehensive review of oyster peptides: Preparation, characterisation and bioactivities

Engineering His-Tagged Senecavirus A for One-Step Purification of Viral Antigens

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