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Microbot Preparation

Microbot Preparation. Joseph Lau and Bobby Wei. Vehicle for “ bacfection ”. Strepavidin and biotin. Streptavidin has one of the strongest non-covalent interactions known to chemistry with the vitamin biotin

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Microbot Preparation

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  1. Microbot Preparation Joseph Lau and Bobby Wei

  2. Vehicle for “bacfection”

  3. Strepavidin and biotin Streptavidin has one of the strongest non-covalent interactions known to chemistry with the vitamin biotin Biotin is a water-soluble B-complex vitamin that can be easily added to antibodies Mechanism: - Biotinylated antibodies latch onto receptor proteins on a bacteria’s surface and also bind to streptavidin-coated nanobots containing cargo

  4. Advantages to nanoparticle system • Many copies of a particular cargo can be carried on a single bacterium • Because bacteria do not need to be transformed or genetically engineered in any way, method is simpler and cheaper • Versatile sizes and types of cargo can be attached inside streptavidin-ated polystyrene nanoparticles

  5. Preparation of the biotinylated antibodies • Monoclonal Antibody C11E9 • Biotinylated-anti-Listeria antibody • Buy from Invitrogen • Make payment • Open package

  6. Preparation of Nanoparticles • Streptavidin or neutravidin-immobilized green (200nm, 40nm) or red (40nm) fluorescent polystyrene particles (1 and 0.5% solids content, respectively) • Purchase from a commercial source (FluoSpheres, Invitrogen, Carlsbad, CA) and were used as supplied by the manufacturer throughout the study

  7. Preparation of microbots • Mix L. monocytogenes with phosphate buffered saline (PBS), Monoclonal Antibody C11E9, and centrifuge 10,000 xg for 5 minutes • Mix bacterium with streptavidinated-nanoparticles and incubate for 15 minutes

  8. The reason we’re doing this: • A lot of projects contain existing technologies that are combined in novel ways. • This novel method of drug delivery and localization of bacteria will involve using the strepavidin-biotin extracellular system as well as ribozymes => iGEM victory

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