Cognate peptide-receptor ligand mapping by directed phage display
<p>Abstract</p> <p>Background</p> <p>A rapid phage display method for the elucidation of cognate peptide specific ligand for receptors is described. The approach may be readily integrated into the interface of genomic and proteomic studies to identify biologically relev...
| Main Authors: | , |
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| Format: | Article |
| Language: | English |
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BMC
2005-06-01
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| Series: | Proteome Science |
| Online Access: | http://www.proteomesci.com/content/3/1/7 |
| _version_ | 1818381564357115904 |
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| author | Stratmann Thomas Kang Angray S |
| author_facet | Stratmann Thomas Kang Angray S |
| author_sort | Stratmann Thomas |
| collection | DOAJ |
| description | <p>Abstract</p> <p>Background</p> <p>A rapid phage display method for the elucidation of cognate peptide specific ligand for receptors is described. The approach may be readily integrated into the interface of genomic and proteomic studies to identify biologically relevant ligands.</p> <p>Methods</p> <p>A gene fragment library from influenza coat protein haemagglutinin (HA) gene was constructed by treating HA cDNA with DNAse I to create 50 – 100 bp fragments. These fragments were cloned into plasmid pORFES IV and in-frame inserts were selected. These in-frame fragment inserts were subsequently cloned into a filamentous phage display vector JC-M13-88 for surface display as fusions to a synthetic copy of gene <it>VIII</it>. Two well characterized antibodies, mAb 12CA5 and pAb 07431, directed against distinct known regions of HA were used to pan the library.</p> <p>Results</p> <p>Two linear epitopes, HA peptide 112 – 126 and 162–173, recognized by mAb 12CA5 and pAb 07431, respectively, were identified as the cognate epitopes.</p> <p>Conclusion</p> <p>This approach is a useful alternative to conventional methods such as screening of overlapping synthetic peptide libraries or gene fragment expression libraries when searching for precise peptide protein interactions, and may be applied to functional proteomics.</p> |
| first_indexed | 2024-12-14T02:36:35Z |
| format | Article |
| id | doaj.art-a46f537807a545e69a3ee663238a456d |
| institution | Directory Open Access Journal |
| issn | 1477-5956 |
| language | English |
| last_indexed | 2024-12-14T02:36:35Z |
| publishDate | 2005-06-01 |
| publisher | BMC |
| record_format | Article |
| series | Proteome Science |
| spelling | doaj.art-a46f537807a545e69a3ee663238a456d2022-12-21T23:20:07ZengBMCProteome Science1477-59562005-06-0131710.1186/1477-5956-3-7Cognate peptide-receptor ligand mapping by directed phage displayStratmann ThomasKang Angray S<p>Abstract</p> <p>Background</p> <p>A rapid phage display method for the elucidation of cognate peptide specific ligand for receptors is described. The approach may be readily integrated into the interface of genomic and proteomic studies to identify biologically relevant ligands.</p> <p>Methods</p> <p>A gene fragment library from influenza coat protein haemagglutinin (HA) gene was constructed by treating HA cDNA with DNAse I to create 50 – 100 bp fragments. These fragments were cloned into plasmid pORFES IV and in-frame inserts were selected. These in-frame fragment inserts were subsequently cloned into a filamentous phage display vector JC-M13-88 for surface display as fusions to a synthetic copy of gene <it>VIII</it>. Two well characterized antibodies, mAb 12CA5 and pAb 07431, directed against distinct known regions of HA were used to pan the library.</p> <p>Results</p> <p>Two linear epitopes, HA peptide 112 – 126 and 162–173, recognized by mAb 12CA5 and pAb 07431, respectively, were identified as the cognate epitopes.</p> <p>Conclusion</p> <p>This approach is a useful alternative to conventional methods such as screening of overlapping synthetic peptide libraries or gene fragment expression libraries when searching for precise peptide protein interactions, and may be applied to functional proteomics.</p>http://www.proteomesci.com/content/3/1/7 |
| spellingShingle | Stratmann Thomas Kang Angray S Cognate peptide-receptor ligand mapping by directed phage display Proteome Science |
| title | Cognate peptide-receptor ligand mapping by directed phage display |
| title_full | Cognate peptide-receptor ligand mapping by directed phage display |
| title_fullStr | Cognate peptide-receptor ligand mapping by directed phage display |
| title_full_unstemmed | Cognate peptide-receptor ligand mapping by directed phage display |
| title_short | Cognate peptide-receptor ligand mapping by directed phage display |
| title_sort | cognate peptide receptor ligand mapping by directed phage display |
| url | http://www.proteomesci.com/content/3/1/7 |
| work_keys_str_mv | AT stratmannthomas cognatepeptidereceptorligandmappingbydirectedphagedisplay AT kangangrays cognatepeptidereceptorligandmappingbydirectedphagedisplay |