Fletcher, S.T., Basketter, D.A. Safety and Environmental Assurance Centre (SEAC), Unilever Colworth, Sharnbrook, Bedfordshire, UK.

This study by researchers at Unilever’s Safety and Environmental Assurance Centre (SEAC) demonstrated how MatTek’s EpiDerm in vitro human skin tissue equivalent was used to identify a number of proteins that could be used as general markers for skin irritation. Scientists at Unilever’s Safety and Environmental Assurance Centre (UK) performed an analysis of EpiDerm™ cultures treated with the known skin irritant sodium lauryl sulphate (SLS) using 2D-gel electrophoresis in order to understand the mechanism of action and thereby identify novel markers of skin irritation. A range of both broad and narrow pH gradient first-dimension gels were run (pH 4-7, 6-11, 4-5, 5-6 and 6-9) consistently followed by 12% SDS-PAGE in the second dimension. Following treatment of EpiDerm with SLS, 67 proteins of interest were identified, of which 8 were selected as interesting: calmodulin-like skin protein, involucrin, epithelial cell marker protein, HS1, per-oxiredoxin 1, serine protease inhibitor, KIAA0117 and ribosomal protein L17. Involucrin was confirmed as being up-regulated by both ELISA and Western blotting. The use of proteomics and EpiDerm human skin tissue equivalents identified a number of proteins which could be used as general markers for skin irritation, and which may in particular be of value for the development of in vitro predictive models.


2D-gel electrophoresis, Allergic reactions, Cytotoxicity, ECVAM, ELISA, EpiDerm, HS1, In vitro alternatives, Involucrin, MTT, MTT assay, Peroxiredoxin 1, Proteomic analysis, Proteomicis, Rapid test system, Serine protease inhibitor, Skin irritation, Skin irritation mechanisms, Sodium lauryl sulphate (SLS), Western blotting

Materials Tested

62387, ALEX1 protein, Actin, Albumin, Arginase, Calmodulin-like skin protein, Cytoskeletal tropomysin, Epithelial cell marker protein, GARS protein, GDP dissociation inhibitor 2, Galectin 7, Gap junction protein, HS1, HSP 27, Heat shock protein 27 kDa, Heat shock protein 60 k Da, Heterogeneous nuclear ribonucleoprotein, High mobility group protein, HsGCN1, Hypothetical protein FLJ21620, Involucrin, JM10 protein, KIAA0117, KIAA1505 protein, Keratin 1, Keratin 17, MarR family, Myosin heavy chain, non muscle form, Myosin heavy polypeptide OR Spectrin, NADH dehydrogenase, Peroxiredoxin 1, Phosphotriesterase related protein, Plectin, Probable mucin DKFZp434C196.1, Prohibitin, Replication factor C, Ribosomal protein L14, Ribosomal protein L17, Serine protease inhibitor, Sestrin 2, Similar to 60s ribosomal protein, Solute carrier family 25, Splicing factor arginine/serine rich, Squamous cell carcinoma antigen 1, T cell receptor delta chain, Transcription regulator, Trophoblast-endothelial activated lymphocyte, Urocortin preprotein, Zinc finger protein 221

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