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Automated Protein Dilutor and Interferometer

Automated Protein Dilutor and Interferometer . Anthony J. Cipolla University of New Hampshire ECE 791/792 Proposal Presentation ECE Faculty Advisor: Dr. Allen Drake CAMIS Project Director: Dr. Thomas Laue Date: October 28, 2010 Project Completion: May 2011. Agenda. Project Introduction

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Automated Protein Dilutor and Interferometer

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  1. Automated Protein Dilutor and Interferometer Anthony J. Cipolla University of New Hampshire ECE 791/792 Proposal Presentation ECE Faculty Advisor: Dr. Allen Drake CAMIS Project Director: Dr. Thomas Laue Date: October 28, 2010 Project Completion: May 2011

  2. Agenda • Project Introduction • Definition • Biotech Overview • Objectives • Budget • Timeline • Diagrams

  3. Project Introduction • Problem: Dilute and measure highly concentrated protein for biotech drug development • Solution: Construct unit to pump out waste, pump in buffer solution, aliquot volumes of 10 µL in 3.5 mL cuvette • Interferometer will be constructed to measure and monitor concentration using the refractive index in order to find the chemical activity coefficient • Center to Advance Molecular Interaction Sciences contracted by MedImmune • MedImmune was first company to launch a mAb for infectious disease in US (1998)

  4. Project Definition Design and construct a bench-top automated protein dilutor and interferometric device to measure and monitor concentration.

  5. Biotech Overview

  6. Biotech vs. Pharmaceutical Drugs Biotech • Antibodies (proteins) expressed by living cells • Harder to develop • Drug function highly sensitive to small supplement variations • Primarily injectables Pharma • Chemically synthesized molecules • Easier to develop • Physical crystallized molecular structure easy to “reverse engineer” • Primarily ingestibles

  7. What is an IgG protein? • Immunoglobin G are antibody molecules • Found in blood • Only antibody that can cross placenta to give immunity fetus • Apparent molar mass in range of 150kg/mol • Formulated in pH 6

  8. Molecular Proximity = Big Issue • Collision distance less than molecular diameter

  9. Bimolecular Collisions • Energy exchanges • Ionic strength effects • pH dependence • Predict protein behavior

  10. Where does dilution take place in drug development process? • Downstream processing • After cells are grown in a soup of growth media, supplements and oxygen • After proteins are filtered from cells • Help scientists: • Process drug for desired injection concentration • Find chemical activity coefficient

  11. Dilution Sequence 1. Remove aliquot into waste container 2. Add aliquot of buffer solution 3. Stir media for 20 seconds 4. Light scattering - Wyatt Dawn HELEOS II 5. Find activity coefficient with interferometer 6. Process repeats fifty times in 12 hours

  12. Dilutor Diagram

  13. Interferometer • A Rayleigh Interferometer • Two beams of light from a single source • Interference pattern viewed by a solid-state camera detector and shown on a television • True molecular weight can be determined based on the fringe displacement • Find the chemical activity coefficient

  14. Interferometer Diagrams

  15. Interferometer Processing • The solid-state television camera will have a resolution suitable for collecting an array of pixels that will be digitally processed • An algorithm will process the array of pixels and find the true molecular weight of the sample

  16. Objectives 1. Program Hamilton dual syringe dilutor 2. Hamilton to Spectrocell communication 3. Retrofit Mini-Motor to drive PTFE mixing shaft 4. Secure mixing motor to cell cuvette 5. Concentration calculation program 6. Interferometer – concentration measurement 7. Solid-state camera to pixels, TV 8. Algorithm to process fringe displacement

  17. Enable Motor Controller Communication from Syringe Dilutor

  18. Budget • Total budget is $10,000 • Dr. Thomas Laue, director of the Center to Advance Molecular Interaction Sciences (CAMIS) at the University of New Hampshire, is contracted by MedImmune, LLC to complete this project

  19. Timeline

  20. Diagrams

  21. Diagrams (continued)

  22. Q & A

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