Peptide Production and Characterization

Chemically synthesized AMP peptide (LDALVKEKKLQGKGPGGPPPK), a scrambled peptide (GKPLGQPGKVPKLDGKEPLAK), and recombinant human (rh)AMP-18 were prepared by GenScript (Piscataway, NJ) as described previously [13 (link)]. Briefly, rhAMP-18 is expressed and purified as a His₆-tagged fusion protein. The coding sequence for full-length human AMP-18 was cloned into an E. coli expression vector, pGSE3, and the expressed protein was purified from 5 L of culture medium by affinity column chromatography. AMP peptide and rhAMP-18 were found to be equally effective in triggering cellular responses as previously reported [13 (link)–15 (link)]. Cell culture medium, fetal bovine serum (FBS), penicillin and streptomycin were obtained from Gibco BRL, Life Technologies (Gaithersburg, MD). Tumor necrosis factor (TNF)-α was purchased from PeproTech (Rocky Hill, NJ), and other reagents from Sigma-Aldrich unless otherwise specified.

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Chen P., Mancini M., Sonis S.T., Fernandez-Martinez J., Liu J., Cohen E.E, & Toback F.G. (2016). A Novel Peptide for Simultaneously Enhanced Treatment of Head and Neck Cancer and Mitigation of Oral Mucositis. PLoS ONE, 11(4), e0152995.

Publication 2016

fetal bovine serum (FBS), penicillin streptomycin Tumor necrosis factor (TNF)-α

Affinity chromatography Cell Cell culture Coding sequence Culture medium E coli Fetal bovine serum Human Penicillin Peptide Protein Protein a Streptomycin Tumor necrosis factor α Vector

Corresponding Organization : Universidad de Oviedo

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Variable analysis

independent variables

Chemically synthesized AMP peptide (LDALVKEKKLQGKGPGGPPPK)
Scrambled peptide (GKPLGQPGKVPKLDGKEPLAK)
Recombinant human (rh)AMP-18

dependent variables

Cellular responses

control variables

Cell culture medium
Fetal bovine serum (FBS)
Penicillin and streptomycin
Tumor necrosis factor (TNF)-α

positive control

AMP peptide and rhAMP-18 were found to be equally effective in triggering cellular responses as previously reported [13 (link)–15 (link)].

negative control

Not explicitly mentioned.

Annotations

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