Abstract
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.
Highlights
Cancer is a leading cause of death, and the cancer incidence and cancer death rate are rising steadily [1]
The amount of bacteriophages M13 required for chemical modification was ampliThe amount of bacteriophages M13 required for chemical modification was amplified fied in cultures of E. coli strain ER2738
Phage modification can contribute to the development of the new molecular imaging probes and techniques for the early detection of cancer; bacteriophages themselves are particles with therapeutic potential
Summary
Cancer is a leading cause of death, and the cancer incidence and cancer death rate are rising steadily [1]. The filamentous bacteriophage, due to its ability to display or bind various molecules on the surface of the capsid and an innate capability to penetrate and traverse tissues and barriers, can be effectively used to deliver drug delivery vehicles for cancer treatment [3]. The mechanism of internalization, intracellular transport, and stability of tumor-targeting filamentous phage M13 inside a eukaryotic cell was described earlier [5]. 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. Results: We performed chemical phage modification using different concentrations of FAM-NHS dye
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