Preparation of recombinant murine tumor necrosis factor-α in Escherichia coli: A rapid method to remove tags from fusion proteins by thrombin-cleavage and ion-exchange chromatography

Tsukamoto, Hiroki; Fukudome, Kenji; Kohara, Jun; Nakatake, Hiroshi; Kimoto, Masao

doi:10.1016/j.pep.2007.07.004

Cited by 12 publications

(9 citation statements)

References 22 publications

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“…Samples were then transferred to corresponding wells on L929 culture plates and incubated for additional 24 h in the presence of actinomycin D at 37 °C. 20,21 Cells were finally assayed for cell viability using the CellTiter Aqueous One Solution reagent (MTS, Promega) directly to the culture as previously described. Residual in vitro bioactivity was determined by normalizing OD values from each sample to the values obtained by cells cultured in the absence of TNF- α .…”

Section: Methodsmentioning

confidence: 99%

Complexation Hydrogels as Oral Delivery Vehicles of Therapeutic Antibodies: An in Vitro and ex Vivo Evaluation of Antibody Stability and Bioactivity

Carrillo-Conde¹,

Brewer²,

Lowman

et al. 2015

Ind. Eng. Chem. Res.

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Oral administration of monoclonal antibodies (mAbs) may enable the localized treatment of infections or other conditions in the gastrointestinal tract (GI) as well as systemic diseases. As with the development of oral protein biotherapeutics, one of the most challenging tasks in antibody therapies is the loss of biological activity due to physical and chemical instabilities. New families of complexation hydrogels with pH-responsive properties have demonstrated to be excellent transmucosal delivery vehicles. This contribution focuses on the design and evaluation of hydrogel carriers that will minimize the degradation and maximize the in vivo activity of anti-TNF-α, a mAb used for the treatment of inflammatory bowel disease (IBD) in the GI tract and systemically for the treatment of rheumatoid arthritis. P(MAA-g-EG) and P(MAA-co-NVP) hydrogels systems were optimized to achieve adequate swelling behavior, which translated into improved protein loading and release at neutral pH simulating the small intestine conditions. Additionally, these hydrogel systems preserve antibody bioactivity upon release resulting in the systemic circulation of an antibody capable of effectively performing its biological function. The compatibility if these hydrogels for mAb bioactivity preservation and release makes them candidates for use as oral delivery systems for therapeutic antibodies.

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Section: Methodsmentioning

confidence: 99%

Complexation Hydrogels as Oral Delivery Vehicles of Therapeutic Antibodies: An in Vitro and ex Vivo Evaluation of Antibody Stability and Bioactivity

Carrillo-Conde¹,

Brewer²,

Lowman

et al. 2015

Ind. Eng. Chem. Res.

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“…The activity of the recombinant proteins was determined using TNF-␣-sensitive L929 fibroblasts as described before (17). L929 cells were treated with serial dilutions of recombinant TNF-␣ for 24 h in the presence of actinomycin D (Sigma-Aldrich, St. Louis, MO, USA, 1 g/ml).…”

Section: Methodsmentioning

confidence: 99%

Development of a Recombinant Xenogeneic Tumor Necrosis Factor Alpha Protein Vaccine To Protect Mice from Experimental Colitis

Wan

Zhang

et al. 2015

Clin. Vaccine Immunol.

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dPrevious studies have highlighted the efficacy of tumor necrosis factor alpha (TNF-␣) inhibitors, including monoclonal antibodies and soluble receptors, in the treatment and management of intestinal bowel disease (IBD). However, because of the immunogenicity of xenogeneic TNF-␣ inhibitors, antidrug antibodies (ADAs) can be triggered after repeated administration. An alternative way to target TNF-␣ is active immunization to elicit the production of high titers of neutralizing antibodies. In this study, we prepared a xenogeneic TNF-␣ protein vaccine and studied the protective effects in experimental colitis models. The xenogeneic TNF-␣ protein vaccine could overcome self-tolerance and induce TNF-␣-specific neutralizing antibody. Moreover, the xenogeneic TNF-␣ protein vaccine could protect mice from acute and chronic colitis induced by dextran sodium sulfate (DSS). One possible explanation for this protective effect is the production of TNF-␣-specific neutralizing antibody, which absorbed the biological activity of mouse TNF-␣ (mTNF-␣) and failed to induce T lymphocyte apoptosis. In summary, use of the xenogeneic TNF-␣ protein vaccine may be a potent therapeutic strategy for IBD.

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“…Anti-TNF-α antibody functional stability was evaluated in a previously described potency assay based on the TNF-α-induced cytotoxicity of L929 murine fibroblast cells (O'Connell et al, 1997;Tsukamoto et al, 2007). In brief, L929 cells (ATCC, Manassas, VA) were cultured in Dulbecco's Modified Eagle Medium (DMEM, Invitrogen, Carlsbad, CA) supplemented with 10% heatinactivated FBS (Valley Biomedical, Winchester, VA), 100 IU/mL penicillin and 10 μg/mL streptomycin (Mediatech, Herndon, VA).…”

Section: Anti-tnf-α Functional Stabilitymentioning

confidence: 99%

“…Bioassay plates were then incubated an additional 24 h in the presence of actinomycin D at 37 1C (O'Connell et al, 1997;Tsukamoto et al, 2007). After incubation, cell viability was analyzed using an MTT assay.…”

Section: Anti-tnf-α Functional Stabilitymentioning

confidence: 99%

Sustained release and stabilization of therapeutic antibodies using amphiphilic polyanhydride nanoparticles

Carrillo-Conde

Darling

Seiler

et al. 2015

Chemical Engineering Science

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Preparation of recombinant murine tumor necrosis factor-α in Escherichia coli: A rapid method to remove tags from fusion proteins by thrombin-cleavage and ion-exchange chromatography

Cited by 12 publications

References 22 publications

Complexation Hydrogels as Oral Delivery Vehicles of Therapeutic Antibodies: An in Vitro and ex Vivo Evaluation of Antibody Stability and Bioactivity

Complexation Hydrogels as Oral Delivery Vehicles of Therapeutic Antibodies: An in Vitro and ex Vivo Evaluation of Antibody Stability and Bioactivity

Development of a Recombinant Xenogeneic Tumor Necrosis Factor Alpha Protein Vaccine To Protect Mice from Experimental Colitis

Sustained release and stabilization of therapeutic antibodies using amphiphilic polyanhydride nanoparticles

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