Expression, Purification, and Characterization of Recombinant Human Interleukin-13 from NS-0 Cells

Cannon-Carlson, Susan; Varnerin, Jeff; Tsarbopoulos, Anthony; Jenh, Chung‐Her; Cox, Mary Ann; Chou, Chuan‐Chu; Connelly, Nancy A.; Zavodny, Paul J.; Tang, John C.-T.

doi:10.1006/prep.1997.0835

Cited by 11 publications

(15 citation statements)

References 30 publications

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“…4). The spectrum is similar to that reported previously for hIL-13 (34,37). The CD spectral features persisted at high temperature (Fig.…”

Section: Biochemical and Biophysical Analysis Of Refolded Hil-13supporting

confidence: 87%

“…This finding resolves an earlier confusion based on molecular modeling (35). Previous reports of the production of recombinant hIL-13 include expression in murine NS-O cells (34) and E. coli (37). In terms of NMR studies, the NS-O expression system would not be amenable to isotopic labeling, and the reported E. coli system produces a two-to fourfold lower yield than that reported here, even in rich media.…”

Section: Discussionsupporting

confidence: 54%

“…hIL-13 has previously been expressed in murine NS-O cells (34). However, in contrast to the predictive model of Bamborough et al (35) which contained one disulfide and two free sulfhydryls, no free sulfhydryls were found.…”

Section: Il-13mentioning

confidence: 87%

“…However, in contrast to the predictive model of Bamborough et al (35) which contained one disulfide and two free sulfhydryls, no free sulfhydryls were found. Far-UV circular dichroism measurements indicated high helical content and bioactivity was found independent of glycosylation (34,36). A recent report described production of hIL-13 in Escherichia coli (37); however, the yield was rather low.…”

Section: Il-13mentioning

confidence: 95%

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Expression, Purification, Refolding, and Characterization of Recombinant Human Interleukin-13: Utilization of Intracellular Processing

Eisenmesser

Kapust

Nawrocki

et al. 2000

Protein Expression and Purification

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“…4). The spectrum is similar to that reported previously for hIL-13 (34,37). The CD spectral features persisted at high temperature (Fig.…”

Section: Biochemical and Biophysical Analysis Of Refolded Hil-13supporting

confidence: 87%

Section: Discussionsupporting

confidence: 54%

Section: Il-13mentioning

confidence: 87%

Section: Il-13mentioning

confidence: 95%

See 2 more Smart Citations

Expression, Purification, Refolding, and Characterization of Recombinant Human Interleukin-13: Utilization of Intracellular Processing

Eisenmesser

Kapust

Nawrocki

et al. 2000

Protein Expression and Purification

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“…All of these make the use of E. coli for large-scale production of rhIL-13 at low cost unrealistic. The production of rhIL-13 in murine NS-O cells is also inefficient, as NS-O cell-derived rhIL-13 exists in both monomeric and trimeric forms (Cannon-Carlson et al, 1998). As the latter form has no biological activity, it must be separated from the biologically active monomeric form of rhIL-13 through a multi-step process, lowering production efficiency and raising costs.…”

Section: Cytokinesmentioning

confidence: 99%

Tobacco, a highly efficient green bioreactor for production of therapeutic proteins

Tremblay

Wang

Jevnikar

et al. 2010

Biotechnology Advances

162

123

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Molecular farming of pharmaceuticals in plants has the potential to provide almost unlimited amounts of recombinant proteins for use in disease diagnosis, prevention or treatment. Tobacco has been and will continue to be a major crop for molecular farming and offers several practical advantages over other crops. It produces significant leaf biomass, has high soluble protein content and is a non-food crop, minimizing the risk of food-chain contamination. This, combined with its flexibility and highly-efficient genetic transformation/regeneration, has made tobacco particularly well suited for plant-based production of biopharmaceutical products. The goal of this review is to provide an update on the use of tobacco for molecular farming of biopharmaceuticals as well the technologies developed to enhance protein production/purification/efficacy. We show that tobacco is a robust biological reactor with a multitude of applications and may hold the key to success in plant molecular farming.

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Mass spectrometric mapping of disulfide bonds in recombinant human interleukin-13

Tsarbopoulos¹,

Varnerin²,

Cannon-Carlson³

et al. 2000

J. Mass Spectrom.

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Interleukin 13 (IL‐13), a member of the α‐helical family of cytokines, has ∼30% primary sequence homology with IL‐4 and shares a common receptor component. The biologically active rhIL‐13 is monomeric and non‐glycosylated, and contains two disulfide bonds as determined by comparative electrospray mass spectrometric (MS) analysis of the protein before and after reduction with dithiothreitol–dithioerythritol. A trypsin‐resistant core peptide of rhIL‐13 was isolated and analyzed by plasma desorption (PD) MS, identifying a disulfide‐linked core peptide. Subsequent digestion of this core peptide by pepsin, followed by PDMS analysis of the resulting cystine‐containing peptic fragments, provided rapid determination of the existing disulfide bonds between cysteine residues 28–56 and 44–70. This disulfide arrangement is similar to that observed for the analogous four internal cysteine residues in hIL‐4. The conservation of disulfide bond arrangements between hIL‐13 and hIL‐4, coupled with their α‐helical structure and sequence homologies, confirms that IL‐13 and IL‐4 are structural homologues. It is also consistent with their reported similarities in biological function and receptor binding kinetics. Copyright © 2000 John Wiley & Sons, Ltd.

show abstract

Expression, Purification, and Characterization of Recombinant Human Interleukin-13 from NS-0 Cells

Cited by 11 publications

References 30 publications

Expression, Purification, Refolding, and Characterization of Recombinant Human Interleukin-13: Utilization of Intracellular Processing

Expression, Purification, Refolding, and Characterization of Recombinant Human Interleukin-13: Utilization of Intracellular Processing

Tobacco, a highly efficient green bioreactor for production of therapeutic proteins

Mass spectrometric mapping of disulfide bonds in recombinant human interleukin-13

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