Design, synthesis, and validation of an in vitro platform peptide-whole cell screening assay using MTT reagent
An in vitro platform to perform peptide screening against different cancer cell lines was designed. The strategy for this screening relied on the design and detection of high-affinity cancer-targeting peptides based on the sequences of NGR and P160. Evaluation of the best binding peptides was perfor...
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| Format: | Article |
| Language: | English |
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Taylor & Francis Group
2017-05-01
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| Series: | Journal of Taibah University for Science |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S1658365516300802 |
| _version_ | 1819070704874684416 |
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| author | Sahar Ahmed Kamaljit Kaur |
| author_facet | Sahar Ahmed Kamaljit Kaur |
| author_sort | Sahar Ahmed |
| collection | DOAJ |
| description | An in vitro platform to perform peptide screening against different cancer cell lines was designed. The strategy for this screening relied on the design and detection of high-affinity cancer-targeting peptides based on the sequences of NGR and P160. Evaluation of the best binding peptides was performed via incubation of the peptide array-bounded cells with MTT reagent, which is reduced to purple formazan in living cells and further quantified using an Elispot and Kodak imager. For proof of concept, a peptide library (132 spots, and 66 different peptides) was designed, synthesized, and screened against different cancer cell lines. The current strategy assists in the identification of positive and negative peptides as well as the relative binding between positive ones. Better binding peptide sequences of the NGR motif were demonstrated to show up to a 2.6-fold increase in CD13+ cell lines with insignificant binding to CD13− ones. Comparable results were observed for P160 peptide sequences, to which different peptides had increased binding, with an up to 3-fold increase relative to the native P160 peptide. Based on our results, new peptide sequences for cancer targeting were identified, and the developed strategy was applied to two different peptide libraries. |
| first_indexed | 2024-12-21T17:10:10Z |
| format | Article |
| id | doaj.art-af9ffce624d9432ea1a9555af42e8e6f |
| institution | Directory Open Access Journal |
| issn | 1658-3655 |
| language | English |
| last_indexed | 2024-12-21T17:10:10Z |
| publishDate | 2017-05-01 |
| publisher | Taylor & Francis Group |
| record_format | Article |
| series | Journal of Taibah University for Science |
| spelling | doaj.art-af9ffce624d9432ea1a9555af42e8e6f2022-12-21T18:56:25ZengTaylor & Francis GroupJournal of Taibah University for Science1658-36552017-05-0111348749610.1016/j.jtusci.2016.10.001Design, synthesis, and validation of an in vitro platform peptide-whole cell screening assay using MTT reagentSahar Ahmed0Kamaljit Kaur1Department of Pharmacognosy and Pharmaceutical Chemistry, College of Pharmacy, Taibah University, Al-Madinah Al-munawarah, P.O. Box 344, 41411, Saudi ArabiaChapman University School of Pharmacy (CUSP), Harry and Diane Rinker Health Science Campus, Chapman University, Irvine, CA 92618-1908, USAAn in vitro platform to perform peptide screening against different cancer cell lines was designed. The strategy for this screening relied on the design and detection of high-affinity cancer-targeting peptides based on the sequences of NGR and P160. Evaluation of the best binding peptides was performed via incubation of the peptide array-bounded cells with MTT reagent, which is reduced to purple formazan in living cells and further quantified using an Elispot and Kodak imager. For proof of concept, a peptide library (132 spots, and 66 different peptides) was designed, synthesized, and screened against different cancer cell lines. The current strategy assists in the identification of positive and negative peptides as well as the relative binding between positive ones. Better binding peptide sequences of the NGR motif were demonstrated to show up to a 2.6-fold increase in CD13+ cell lines with insignificant binding to CD13− ones. Comparable results were observed for P160 peptide sequences, to which different peptides had increased binding, with an up to 3-fold increase relative to the native P160 peptide. Based on our results, new peptide sequences for cancer targeting were identified, and the developed strategy was applied to two different peptide libraries.http://www.sciencedirect.com/science/article/pii/S1658365516300802PeptidesCancer targeting peptidesPeptide arraySpot synthesis |
| spellingShingle | Sahar Ahmed Kamaljit Kaur Design, synthesis, and validation of an in vitro platform peptide-whole cell screening assay using MTT reagent Journal of Taibah University for Science Peptides Cancer targeting peptides Peptide array Spot synthesis |
| title | Design, synthesis, and validation of an in vitro platform peptide-whole cell screening assay using MTT reagent |
| title_full | Design, synthesis, and validation of an in vitro platform peptide-whole cell screening assay using MTT reagent |
| title_fullStr | Design, synthesis, and validation of an in vitro platform peptide-whole cell screening assay using MTT reagent |
| title_full_unstemmed | Design, synthesis, and validation of an in vitro platform peptide-whole cell screening assay using MTT reagent |
| title_short | Design, synthesis, and validation of an in vitro platform peptide-whole cell screening assay using MTT reagent |
| title_sort | design synthesis and validation of an in vitro platform peptide whole cell screening assay using mtt reagent |
| topic | Peptides Cancer targeting peptides Peptide array Spot synthesis |
| url | http://www.sciencedirect.com/science/article/pii/S1658365516300802 |
| work_keys_str_mv | AT saharahmed designsynthesisandvalidationofaninvitroplatformpeptidewholecellscreeningassayusingmttreagent AT kamaljitkaur designsynthesisandvalidationofaninvitroplatformpeptidewholecellscreeningassayusingmttreagent |