OBJECTIVETo 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.

METHODSHuman 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.

RESULTSAfter 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.

CONCLUSIONSThe 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.

Amino Acid Sequence ; Base Sequence ; Enzyme-Linked Immunosorbent Assay ; Fluorescent Antibody Technique ; Humans ; Osteoblasts ; Peptide Library ; Peptides ; genetics ; Sequence Analysis, DNA ; Sequence Analysis, Protein ; Titanium