In planta production of ELPylated spidroin-based proteins results in non-cytotoxic biopolymers

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• 作者：Valeska Hauptmann (1)
  Matthias Menzel (2)
  Nicola Weichert (1)
  Kerstin Reimers (3)
  Uwe Spohn (2)
  Udo Conrad (1)
  
  1. Leibniz Institute of Plant Genetics and Crop Plant Research ; Corrensstrasse 3 ; 06466 ; Stadt Seeland ; OT Gatersleben ; Germany
  2. Fraunhofer Institute for Mechanics of Materials ; Walter-H眉lse-Strasse 1 ; 06120 ; Halle/Saale ; Germany
  3. Department of Plastic ; Hand and Reconstructive Surgery ; Hannover Medical School ; Podbielskistr. 380 ; 30659 ; Hannover ; Germany
  
• 关键词：Spider silk ; Immunogenicity ; Synthetic spidroin ; Biomaterial ; Biocompatibility ; Atomic force microscopy
• 刊名：BMC Biotechnology
• 出版年：2015
• 出版时间：December 2015
• 年：2015
• 卷：15
• 期：1
• 全文大小：2,530 KB
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• 刊物类别：Chemistry and Materials Science
• 刊物主题：Biotechnology
  Life Sciences
  Plant Breeding/Biotechnology
  Stem Cells
  Transgenics
  
• 出版者：BioMed Central
• ISSN：1472-6750

文摘

Background Spider silk is a tear-resistant and elastic biopolymer that has outstanding mechanical properties. Additionally, exiguous immunogenicity is anticipated for spider silks. Therefore, spider silk represents a potential ideal biomaterial for medical applications. All known spider silk proteins, so-called spidroins, reveal a composite nature of silk-specific units, allowing the recombinant production of individual and combined segments. Results In this report, a miniaturized spidroin gene, named VSO1 that contains repetitive motifs of MaSp1 has been synthesized and combined to form multimers of distinct lengths, which were heterologously expressed as elastin-like peptide (ELP) fusion proteins in tobacco. The elastic penetration moduli of layered proteins were analyzed for different spidroin-based biopolymers. Moreover, we present the first immunological analysis of synthetic spidroin-based biopolymers. Characterization of the binding behavior of the sera after immunization by competitive ELISA suggested that the humoral immune response is mainly directed against the fusion partner ELP. In addition, cytocompatibility studies with murine embryonic fibroblasts indicated that recombinant spidroin-based biopolymers, in solution or as coated proteins, are well tolerated. Conclusion The results show that spidroin-based biopolymers can induce humoral immune responses that are dependent on the fusion partner and the overall protein structure. Furthermore, cytocompatibility assays gave no indication of spidroin-derived cytotoxicity, suggesting that recombinant produced biopolymers composed of spider silk-like repetitive elements are suitable for biomedical applications.