کد مقاله کد نشریه سال انتشار مقاله انگلیسی نسخه تمام متن
4752102 1415992 2017 12 صفحه PDF دانلود رایگان
عنوان انگلیسی مقاله ISI
Regular article“In cell” biotinylation and immobilization of hBMP-2 (human Bone Morphogenetic Protein 2) on polymeric surfaces
موضوعات مرتبط
مهندسی و علوم پایه مهندسی شیمی بیو مهندسی (مهندسی زیستی)
پیش نمایش صفحه اول مقاله
Regular article“In cell” biotinylation and immobilization of hBMP-2 (human Bone Morphogenetic Protein 2) on polymeric surfaces
چکیده انگلیسی


- Directed “in cell” biotinyliosis of the Bone Morphogenetic Protein 2 (BMP-2).
- Immobilization of BMP-2 on polymeric surfaces used in orthopedics.
- Differentiation of stem cells to osteoblasts on the bio-functionalized surfaces.

Bone Morphogenetic Protein 2 (BMP-2) is a very well-known factor that induces bone formation in vitro (Yamaguchi et al., 1991) [1] and in vivo (Wang et al., 1990) [2] and is widely used in orthopedics for the reconstitution of bone tissue. A full description of a purification and refolding (Long et al., 2006; Von Einem et al., 2010) [3,4] protocol for the “in cell” biotinylation of hBMP-2 is given in this paper: cloning of the mature hBMP-2 gene in vector pAN5 for the integration of the biotinylation sequence into the N-terminus and subsequently in vector pET29c for simultaneous overproduction and biotinylation of the protein. The biotinylation sequence codes for the specific 14mer - peptide with the unique recognizable lysine for biotin covalent interaction, ensuring the “in cell” biotinylation of hBMP-2 without functional distortion. In addition, the targeted biotinylation process does not affect the osteogenic properties of the protein when free in solution or after its functional immobilization onto “bone cement” (Vaishya et al., 2013) [5] materials. This is supported by the observation of the enhancement of differentiation of mesenchymal cells into osteoblasts on the “bio-functionalized” polymeric surfaces verified by the alizarin red staining and the alkaline phosphatase assay.

ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Biochemical Engineering Journal - Volume 123, 15 July 2017, Pages 1-12
نویسندگان
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