Sterilized Recombinant Spider Silk Fibers of Low Pyrogenicity

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2010
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Hedhammar, My
Bramfeldt, Hanna
Widhe, Mona
Askarieh, Glareh
Nordling, Kerstin
Johansson, Jan
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Biomacromolecules. American Chemical Society (ACS). 2010, 11(4), pp. 953-959. ISSN 1525-7797. eISSN 1526-4602. Available under: doi: 10.1021/bm9014039
Zusammenfassung

We have recently shown that it is possible to recombinantly produce a miniature spider silk protein, 4RepCT, that spontaneously self-assembles into mechanically stable macroscopic fibers (Stark, M.; Grip, S.; Rising, A.; Hedhammar, M.; Engstrom, W.; Hjalm, G.; Johansson, J. Macroscopic fibers self-assembled from recombinant miniature spider silk proteins. Biomacromolecules2007, 8 (5), 1695−1701). When produced as a soluble fusion protein (with thioredoxin) in Escherichia coli, the spider silk protein can be subjected to several purification steps without aggregating. Here, combined purification and endotoxin removal is achieved using a simple cell wash procedure, protein affinity purification, and LPS depletion. No toxic chemicals were included in the process and the protein retained its ability to self-assemble into fibers. With this method, fibers with pyrogenicity corresponding to less than 1 EU/mg could be recovered. Moreover, the fibers could be sterilized through autoclaving with retained morphology, structure, and mechanical properties. This implies that this recombinant silk is suitable for usage as biomaterial, which is further supported by data showing that the fibers allow growth of human primary fibroblasts.

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ISO 690HEDHAMMAR, My, Hanna BRAMFELDT, Teodora BARIS, Mona WIDHE, Glareh ASKARIEH, Kerstin NORDLING, Sonja von AULOCK, Jan JOHANSSON, 2010. Sterilized Recombinant Spider Silk Fibers of Low Pyrogenicity. In: Biomacromolecules. American Chemical Society (ACS). 2010, 11(4), pp. 953-959. ISSN 1525-7797. eISSN 1526-4602. Available under: doi: 10.1021/bm9014039
BibTex
@article{Hedhammar2010-04-12Steri-52706,
  year={2010},
  doi={10.1021/bm9014039},
  title={Sterilized Recombinant Spider Silk Fibers of Low Pyrogenicity},
  number={4},
  volume={11},
  issn={1525-7797},
  journal={Biomacromolecules},
  pages={953--959},
  author={Hedhammar, My and Bramfeldt, Hanna and Baris, Teodora and Widhe, Mona and Askarieh, Glareh and Nordling, Kerstin and Aulock, Sonja von and Johansson, Jan}
}
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