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Chinese Journal of Stomatology ; (12): 438-441, 2015.
Article in Chinese | WPRIM | ID: wpr-294687

ABSTRACT

<p><b>OBJECTIVE</b>To explore the experimental methods that the phage peptide library technology screening human osteoblast specificity polypeptide, which will provide the basis of the experiment of the Ti surface biolization modification.</p><p><b>METHODS</b>Human calvarial osteoblasts were used as the target cells for whole-cell biopanning from a 12-mer peptide phage-display library. Cell eluent and cell lysis buffer were cultivate and count respectively after washing. Then the target cells were analyzed by enzyme-linked immunosorbent assay (ELISA) and immunofluorescence detection to authenticate the positive phage clones by human gingival fibroblast as the absorber cells. The positive phage clones were deduced by DNA sequencing.</p><p><b>RESULTS</b>After four rounds of screening, twenty-two positive phage clones were found out from randomly selected phage monoclonals, whose single-strand DNA were extracted and sequenced. Amino acid sequence of the highest frequency peptide was MGWSWWPETWPM.</p><p><b>CONCLUSIONS</b>The specific peptide against human osteoblasts can be obtained from a phage-display peptide library for use as a new research approach and experimental basis of the biolization modification of the titanium surface.</p>


Subject(s)
Humans , Amino Acid Sequence , Base Sequence , Enzyme-Linked Immunosorbent Assay , Fluorescent Antibody Technique , Osteoblasts , Peptide Library , Peptides , Genetics , Sequence Analysis, DNA , Sequence Analysis, Protein , Titanium
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