Removal of Endotoxin from Recombinant Protein Preparations

Liu, Shigui; Tobias, Rowel B.; McClure, Shannon; Styba, Garth; Shi, Qing; Jackowski, George

doi:10.1016/s0009-9120(97)00049-0

Cited by 306 publications

(235 citation statements)

References 12 publications

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“…In the human body, the LPS, also known as endotoxin, is a potent activator of inflammatory responses as it promotes the release of several of pro-inflammatory cytokines, which may cause high fever, severe tissue damage and death (Erridge et al 2002). Therefore, many procedures for removal of LPS from recombinant protein preparations have been developed and adapted to specific processes to obtain products with different levels of purity (Magalhaes et al 2007;Liu et al 1997;Petsch and Anspach 2000). However, elimination of LPS requires considerable efforts and adds significant cost to downstream purification steps of proteins.…”

Section: Discussionmentioning

confidence: 99%

Production of functional inclusion bodies in endotoxin-free Escherichia coli

Rueda

Cano‐Garrido

Mamat

et al. 2014

Appl Microbiol Biotechnol

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Section: Discussionmentioning

confidence: 99%

Production of functional inclusion bodies in endotoxin-free Escherichia coli

Rueda

Cano‐Garrido

Mamat

et al. 2014

Appl Microbiol Biotechnol

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“…Polyhistidine-tagged TRAIL was purified from Escherichia coli by using standard nickel affinity chromatography as per the manufacturer (Qiagen, Chatsworth, CA). Preparations were made endotoxin-free by sequential detergent extraction (Triton X-114) (29,30). Detergent was removed from the final preparation by using Bio-Rad SM-2 beads (30).…”

Section: Methodsmentioning

confidence: 99%

Combined effect of tumor necrosis factor-related apoptosis-inducing ligand and ionizing radiation in breast cancer therapy

Chinnaiyan

Prasad

Hamstra

et al. 2000

Proc. Natl. Acad. Sci. U.S.A.

480

323

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Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) is a potent endogenous activator of the cell death pathway and functions by activating the cell surface death receptors 4 and 5 (DR4 and DR5). TRAIL is nontoxic in vivo and preferentially kills neoplastically transformed cells over normal cells by an undefined mechanism. Radiotherapy is a common treatment for breast cancer as well as many other cancers. Here we demonstrate that ionizing radiation can sensitize breast carcinoma cells to TRAIL-induced apoptosis. This synergistic effect is p53-dependent and may be the result of radiation-induced up-regulation of the TRAIL-receptor DR5. Importantly, TRAIL and ionizing radiation have a synergistic effect in the regression of established breast cancer xenografts. Changes in tumor cellularity and extracellular space were monitored in vivo by diffusion-weighted magnetic resonance imaging (diffusion MRI), a noninvasive technique to produce quantitative images of the apparent mobility of water within a tissue. Increased water mobility was observed in combined TRAIL-and radiationtreated tumors but not in tumors treated with TRAIL or radiation alone. Histological analysis confirmed the loss of cellularity and increased numbers of apoptotic cells in TRAIL-and radiationtreated tumors. Taken together, our results provide support for combining radiation with TRAIL to improve tumor eradication and suggest that efficacy of apoptosis-inducing cancer therapies may be monitored noninvasively, using diffusion MRI.

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“…Eluted proteins were subsequently dialyzed against cPBS, and endotoxins were measured by end-point chromogenic Limulus amebocyte lysate test (Charles River Laboratories). If necessary, further removal of endotoxin was achieved by Triton X-114 phase separation (63). All preparations contained Ͻ0.015 EU/ml.…”

Section: Recombinant Carp Tnf-␣mentioning

confidence: 99%

Receptor-Mediated and Lectin-Like Activities of Carp (Cyprinus carpio) TNF-α

Forlenza

Magez²,

Scharsack³

et al. 2009

The Journal of Immunology

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Functional characterization of TNF-α in species other than mammalian vertebrates is limited, and TNF-α has been studied in a limited number of fish species, primarily in vitro using recombinant proteins. Studies on TNF-α from different fish species so far pointed to several inconsistencies, in particular with respect to some receptor-mediated activities of fish TNF-α, such as the ability to directly activate phagocytes. In the present study a comprehensive analysis of in vitro as well as in vivo biological activities of two isoforms of carp TNF-α was performed. Our results show that carp TNF-α directly primes carp phagocytes and indirectly promotes typical receptor-mediated activities such as phagocyte activation by acting via endothelial cells. Additionally, for the first time in nonmammalian vertebrate species, the lectin-like activity of fish TNF-α homologs was investigated. Our results show an evolutionary conservation of function of this receptor-independent activity of TNF-α not only in cyprinid fish, but also in perciform and salmonid fish. The role of TNF-α in vivo, during infections of carp with the blood parasite Trypanoplasma borreli, was examined using three fundamentally different but complementary approaches: (1) inhibition of TNF-α expression, (2) overexpression of TNF-α, and (3) inhibition of shedding of membrane-bound TNF-α. Our results show that, also in fish, a tight regulation of TNF-α expression is important, since depletion or excess of TNF-α can make an important difference to survival of infection. Finally, we demonstrate a crucial protective role for membrane-bound TNF-α, which has a yet unexploited function in fish.

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Removal of Endotoxin from Recombinant Protein Preparations

Cited by 306 publications

References 12 publications

Production of functional inclusion bodies in endotoxin-free Escherichia coli

Production of functional inclusion bodies in endotoxin-free Escherichia coli

Combined effect of tumor necrosis factor-related apoptosis-inducing ligand and ionizing radiation in breast cancer therapy

Receptor-Mediated and Lectin-Like Activities of Carp (Cyprinus carpio) TNF-α

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