Discipline: Biological Sciences
Subcategory: Cell and Molecular Biology
Alexandria Brooks - Alabama State University
Co-Author(s): Carrie Sanders, Rana Singleton, and Alain Bopda-Waffo
It is known that Au can be used in the treatment of cancer in nano form as a probe to both target and treat cancerous tissues. And although Au itself can be used in the treatment, an anti-cancer biodrug can be coated onto the Au. However, two problems exist. One, nanoparticles tend to aggregate in vivo and are cleared by the immune system. The only way to prevent aggregation is to keep the nanoparticles separated in vivo. The second problem arises with the nature of biodrugs themselves. Biodrugs are made from the same biomolecules that make up the body, as such; they are also subjected to the same enzymes that degrade biomolecules in the body. This leads to indiscriminate distribution, degradation, and a risk of under-medicating. To compensate, a larger dose of the biodrug is given; however, toxicity becomes the risk. Since it is known that certain peptides (nano-tags) bind Au, we hypothesize that displaying these nano-tags on the surface of our bacteriophage Qβ, and allowing them to bind Au will prevent aggregation. This Au can then be coated with an anti-cancer biodrug, and the Au will convey protection to the biodrug. To achieve this, the genes of three gold-binding peptides: Au0 (LKAHLPPSRLPS), Au1 (VSGSSPDS), and Au2 (TGTSVLIATPYV) were inserted separately into the genome of Qβ at the end of the A1 gene. The resulting recombinant phages, pQβAu0, pQβAu1 and pQβAu2, were transformed with HB 101 E. coli. The plaque assay provided the titer and phage morphology, and RT-PCR confirmed the tag gene size for each construct. A binding assay allowed different concentrations of Au to bind to the recombinant phage. Confirmation and visualization of the phage-nanoparticle complex was verified via Transmission Electron Microscopy (TEM). The next phase is to focus on coating the Au nanoparticles with chemotherapeutic biodrugs.
Funder Acknowledgement(s): NIS
Faculty Advisor: Alain Bopda-Waffo, email@example.com
Role: Transformation; PCR; Plaque assay; gel electrophoresis; amplification