Team:Munich/Results

Phactory

Results

First Blick of Phactory


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BIG BROTHER IS WATCHING YOU

Optimizing a Cell-Free System

Cultivation

Lysis

Processing

Quality Control

Bacteriophage Assembly

Home-Made Systems Can Be Compared To Commercial Systems

Cell-Free Systems Allow Host Independent Bacteriophage Assembly

Quality Control Of Bacteriophages

Assembly Of Clinically Relevant Bacteriophages

Sequ-Into Complements Continuous Engineering Cycles in Phactory

Modular Bacteriophage Composition

Isolation

Phenol-Chloroform Precipitation Achieves High Titers

Encapsulation

Bacteriophages Encapsulated In Alginate Can Withstand Gastric Acid

Phactory yields phages with toxicity levels that allow oral administration to the patient. However, oral delivery requires protection of the phages from rapid degradation in the acidic gastric juice, while direct intravenous application requires additional purification steps. To overcome these hurdles, we prototyped two 3D-printed fluidic devices that can be assembled for less than $5. For oral application, we built a nozzle to encapsulate the phages in monodisperse calcium-alginate microspheres protecting them in the stomach. The alginate solution, ejected from a dispenser needle with a syringe pump, was sheared off by a parallel stream of air. Our results show that after 1 hour incubation in simulated gastric fluid, active phages are successfully released in simulated intestinal fluid. For intravenous administration, we can purify the bacteriophages from the remaining cell-extract via fractionation in a pressure-driven size-exclusion filter system. Additionally, we built microfluidic hardware for our human practice project OraColi.

- Alginat für lokale pH wert erhöhung im magen und auflösung in chelatoren(darm), da Ca2+ Ionen quervernetzen

- Enkapsulierung durch Co-Flow System. Druckluft schert Tropfen von Spritzennadel ab, Düse == Hardware  Quervernetzung 1h in CaCl2 Lsg  Waschen  Verifikation durch Mikroskopie  Fertig für Einsatz (insg. Ca 1.5-2h)

Experimente 1. 3D Modell von SYBR Gold gelabelten Phagen im Droplet durch z-Stack (BF/GFP) 2. Darkfield Bilder von gelabelten Droplets (bild) 3. Droplet Zusammensetzung (Viskosität zweier Alginate) 4. Droplet Größen bei Flussrate und Druck 5. Verhalten im Magen bzw. saurem Millieu (pH: 1) und Pepsin (1h @37°C, 10h @ RT) -> Phagen Degradation (barplot) 6. Verhalten im Darm bzw. pH 7 und Pankreatin (2h @ 37°C) ->Phagen relase über Zeit (plot)

References

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