Modification of a Tumor-Targeting Bacteriophage for Potential Diagnostic Applications
Background: Tumor-targeting bacteriophages can be used as a versatile new platform for the delivery of diagnostic imaging agents and therapeutic cargo. This became possible due to the development of viral capsid modification method. Earlier in our laboratory and using phage display technology, phage...
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MDPI AG
2021-10-01
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| Series: | Molecules |
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| Online Access: | https://www.mdpi.com/1420-3049/26/21/6564 |
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| author | Maya Alexandrovna Dymova Yaroslav Alexandrovich Utkin Maria Denisovna Dmitrieva Elena Vladimirovna Kuligina Vladimir Alexandrovich Richter |
| author_facet | Maya Alexandrovna Dymova Yaroslav Alexandrovich Utkin Maria Denisovna Dmitrieva Elena Vladimirovna Kuligina Vladimir Alexandrovich Richter |
| author_sort | Maya Alexandrovna Dymova |
| collection | DOAJ |
| description | Background: Tumor-targeting bacteriophages can be used as a versatile new platform for the delivery of diagnostic imaging agents and therapeutic cargo. This became possible due to the development of viral capsid modification method. Earlier in our laboratory and using phage display technology, phages to malignant breast cancer cells MDA-MB 231 were obtained. The goal of this study was the optimization of phage modification and the assessment of the effect of the latter on the efficiency of phage particle penetration into MDA-MB 231 cells. Methods: In this work, we used several methods, such as chemical phage modification using FAM-NHS ester, spectrophotometry, phage amplification, sequencing, phage titration, flow cytometry, and confocal microscopy. Results: We performed chemical phage modification using different concentrations of FAM-NHS dye (0.5 mM, 1 mM, 2 mM, 4 mM, 8 mM). It was shown that with an increase of the modification degree, the phage titer decreases. The maximum modification coefficient of the phage envelope with the FAM–NHS dye was observed with 4 mM modifying agent and had approximately 804,2 FAM molecules per phage. Through the immunofluorescence staining and flow cytometry methods, it was shown that the modified bacteriophage retains the ability to internalize into MDA-MB-231 cells. The estimation of the number of phages that could have penetrated into one tumor cell was conducted. Conclusions: Optimizing the conditions for phage modification can be an effective strategy for producing tumor-targeting diagnostic and therapeutic agents, i.e., theranostic drugs. |
| first_indexed | 2024-03-10T05:56:05Z |
| format | Article |
| id | doaj.art-b09de5a7df9e4c7994f631e3c7696a02 |
| institution | Directory Open Access Journal |
| issn | 1420-3049 |
| language | English |
| last_indexed | 2024-03-10T05:56:05Z |
| publishDate | 2021-10-01 |
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| record_format | Article |
| series | Molecules |
| spelling | doaj.art-b09de5a7df9e4c7994f631e3c7696a022023-11-22T21:23:13ZengMDPI AGMolecules1420-30492021-10-012621656410.3390/molecules26216564Modification of a Tumor-Targeting Bacteriophage for Potential Diagnostic ApplicationsMaya Alexandrovna Dymova0Yaroslav Alexandrovich Utkin1Maria Denisovna Dmitrieva2Elena Vladimirovna Kuligina3Vladimir Alexandrovich Richter4Institute of Chemical Biology and Fundamental Medicine, Siberian Branch of the Russian Academy of Sciences, 630090 Novosibirsk, RussiaInstitute of Chemical Biology and Fundamental Medicine, Siberian Branch of the Russian Academy of Sciences, 630090 Novosibirsk, RussiaInstitute of Chemical Biology and Fundamental Medicine, Siberian Branch of the Russian Academy of Sciences, 630090 Novosibirsk, RussiaInstitute of Chemical Biology and Fundamental Medicine, Siberian Branch of the Russian Academy of Sciences, 630090 Novosibirsk, RussiaInstitute of Chemical Biology and Fundamental Medicine, Siberian Branch of the Russian Academy of Sciences, 630090 Novosibirsk, RussiaBackground: Tumor-targeting bacteriophages can be used as a versatile new platform for the delivery of diagnostic imaging agents and therapeutic cargo. This became possible due to the development of viral capsid modification method. Earlier in our laboratory and using phage display technology, phages to malignant breast cancer cells MDA-MB 231 were obtained. The goal of this study was the optimization of phage modification and the assessment of the effect of the latter on the efficiency of phage particle penetration into MDA-MB 231 cells. Methods: In this work, we used several methods, such as chemical phage modification using FAM-NHS ester, spectrophotometry, phage amplification, sequencing, phage titration, flow cytometry, and confocal microscopy. Results: We performed chemical phage modification using different concentrations of FAM-NHS dye (0.5 mM, 1 mM, 2 mM, 4 mM, 8 mM). It was shown that with an increase of the modification degree, the phage titer decreases. The maximum modification coefficient of the phage envelope with the FAM–NHS dye was observed with 4 mM modifying agent and had approximately 804,2 FAM molecules per phage. Through the immunofluorescence staining and flow cytometry methods, it was shown that the modified bacteriophage retains the ability to internalize into MDA-MB-231 cells. The estimation of the number of phages that could have penetrated into one tumor cell was conducted. Conclusions: Optimizing the conditions for phage modification can be an effective strategy for producing tumor-targeting diagnostic and therapeutic agents, i.e., theranostic drugs.https://www.mdpi.com/1420-3049/26/21/6564MDA-MB 231 cellstumor-targeting bacteriophageschemical modification of bacteriophagesFAM–NHStheranostic drugs |
| spellingShingle | Maya Alexandrovna Dymova Yaroslav Alexandrovich Utkin Maria Denisovna Dmitrieva Elena Vladimirovna Kuligina Vladimir Alexandrovich Richter Modification of a Tumor-Targeting Bacteriophage for Potential Diagnostic Applications Molecules MDA-MB 231 cells tumor-targeting bacteriophages chemical modification of bacteriophages FAM–NHS theranostic drugs |
| title | Modification of a Tumor-Targeting Bacteriophage for Potential Diagnostic Applications |
| title_full | Modification of a Tumor-Targeting Bacteriophage for Potential Diagnostic Applications |
| title_fullStr | Modification of a Tumor-Targeting Bacteriophage for Potential Diagnostic Applications |
| title_full_unstemmed | Modification of a Tumor-Targeting Bacteriophage for Potential Diagnostic Applications |
| title_short | Modification of a Tumor-Targeting Bacteriophage for Potential Diagnostic Applications |
| title_sort | modification of a tumor targeting bacteriophage for potential diagnostic applications |
| topic | MDA-MB 231 cells tumor-targeting bacteriophages chemical modification of bacteriophages FAM–NHS theranostic drugs |
| url | https://www.mdpi.com/1420-3049/26/21/6564 |
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