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AssigneeEidgenossische Technische Hochschule Zurich
InventorsJeffrey A. Hubbell, Jason C. Schense, Shelly E. Sakiyama
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Heparin-binding regions of several proteins, such as neural cell adhesion molecule, fibronectin, laminin, midkine, and anti-thrombin III have been shown to promote neurite extension on two-dimensional surfaces. The effect of heparin-binding peptides on neurite extension through three-dimensional matrices was investigated by culturing embryonic chick dorsal root ganglia (DRG) within fibrin gels containing chemically attached heparin-binding peptide (HBP). The length of neurites within fibrin gels containing cross-linked HBP was increased by more than 70% over extension through fibrin gels containing no peptide. The HBP sequence of antithrombin III was incorporated into the fibrin gel as the C-terminal domain of a bidomain, chimeric peptide; the N-terminal second domain of this peptide contained the α2-plasmin inhibitor substrate for Factor XIIIa. Factor XIIIa, a transglutaminase, was used to chemically attach the HBP-containing chimeric peptide to the fibrin gels during polymerization. The amount of HBP cross-linked into the fibrin gels was determined, after degradation by plasmin using gel permeation chromatography, to be approximately 8 moles of peptide per mole fibrinogen. A peptide (HBP), where the cross-linking glutamine was replaced with glycine, showed no increase in extension in comparison with fibrin gels. The addition of heparin to the gel precursors resulted in no increase in neurite extension in comparison with fibrin gels. HBPs promote neurite extension by binding to cell surface proteoglycans on the DRG.
A protein matrix comprising fibrin having at least one peptide in a neurite growth and extension promoting amount, when said peptide is cross-linked to said fibrin.
A protein matrix comprising at least one peptide selected from the group consisting of HAV, IKVAV, RNIAEIIKDI, YIGSR, DGEA, a combination thereof, a combination of peptide fragments comprising HAV, IKVAV, RNIAEIIKDI, YIGSR, and DGEA, or a fusion peptide comprising said peptides.
A fibrin gel matrix comprising a mixture of peptides cross-linked thereto, wherein said mixture of peptides comprise IKVAV, RGD, YIGSR and RNIAEIIKDI, wherein said fibrin gel matrix is essentially HAV-free.
A protein gel comprising a neurite extension promoting amount of a ratio of peptide HAV and peptide YIGSR of about 1:3.
A protein gel comprising a neurite extension promoting amount of a ratio of peptide HAV and peptide RNIAEIIKDI of about 1:3.
A protein gel comprising a neurite extension promoting ratio of a first peptide RGD and a second peptide YIGSR.
A protein gel comprising a neurite extension promoting ratio of a peptide RGD and a peptide DEGEA of about 1:3.
A protein gel comprising a neurite extension promoting ratio of a peptide RGD, a peptide YIGSR, and a peptide RNIAEIIKDI of about 1:1:1.
A fibrin gel comprising a neurite extension promoting ratio of a mixture of peptides, wherein said peptides are cross-linked to said fibrin gel.
A protein gel comprising other than fibrin, wherein said protein gel has cross-linked thereto a mixture of peptides at a neurite extension promoting ratio.
A bi-domain peptide comprising a first domain of a heparin-binding domain and a second domain consisting essentially of a Factor XIIIa substrate or a bioactive peptide.
The bi-domain peptide of claim 25 wherein the first domain is further defined as a peptide KβAFAKLAARLYRKA, YKKIIKKL, KHKGRDVILKKDVR, or a mixture thereof.
Fibrin comprising a heparin binding domain from ATIII.
Fibrin comprising a heparin binding domain peptide that elutes from a heparin affinity column at [NaCl]>0.34 Mol.
A protein gel comprising a chemically bound protease inhibitor.
A protein gel comprising a peptide having a protease binding site covalently incorporated to said gel.
A three-dimensional matrix capable of supporting neural cell extension comprising a neurite cell extension promoting amount of a mixture of peptides, wherein each of said peptides comprise a sequence that defines a heparin-binding sequence.
A chimeric peptide comprising:
Dependencies are inferred from the claim text.