Automated Protein Dilutor and Interferometer
description
Transcript of Automated Protein Dilutor and Interferometer
![Page 1: Automated Protein Dilutor and Interferometer](https://reader035.fdocuments.us/reader035/viewer/2022062813/56816696550346895dda7dd1/html5/thumbnails/1.jpg)
Automated Protein Dilutor and Interferometer
Anthony J. CipollaUniversity of New HampshireECE 791/792 Proposal PresentationECE Faculty Advisor: Dr. Allen DrakeCAMIS Project Director: Dr. Thomas LaueDate: October 28, 2010Project Completion: May 2011
![Page 2: Automated Protein Dilutor and Interferometer](https://reader035.fdocuments.us/reader035/viewer/2022062813/56816696550346895dda7dd1/html5/thumbnails/2.jpg)
Project Introduction Definition Biotech Overview Objectives Budget Timeline Diagrams
Agenda
![Page 3: Automated Protein Dilutor and Interferometer](https://reader035.fdocuments.us/reader035/viewer/2022062813/56816696550346895dda7dd1/html5/thumbnails/3.jpg)
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)
Project Introduction
![Page 4: Automated Protein Dilutor and Interferometer](https://reader035.fdocuments.us/reader035/viewer/2022062813/56816696550346895dda7dd1/html5/thumbnails/4.jpg)
Design and construct a bench-top automated protein dilutor and interferometric device to
measure and monitor concentration.
Project Definition
![Page 5: Automated Protein Dilutor and Interferometer](https://reader035.fdocuments.us/reader035/viewer/2022062813/56816696550346895dda7dd1/html5/thumbnails/5.jpg)
Biotech Overview
![Page 6: Automated Protein Dilutor and Interferometer](https://reader035.fdocuments.us/reader035/viewer/2022062813/56816696550346895dda7dd1/html5/thumbnails/6.jpg)
Biotech Antibodies (proteins)
expressed by living cells Harder to develop Drug function highly
sensitive to small supplement variations
Primarily injectables
Biotech vs. Pharmaceutical Drugs
Pharma • Chemically synthesized
molecules• Easier to develop• Physical crystallized
molecular structure easy to “reverse engineer”
• Primarily ingestibles
![Page 7: Automated Protein Dilutor and Interferometer](https://reader035.fdocuments.us/reader035/viewer/2022062813/56816696550346895dda7dd1/html5/thumbnails/7.jpg)
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
![Page 8: Automated Protein Dilutor and Interferometer](https://reader035.fdocuments.us/reader035/viewer/2022062813/56816696550346895dda7dd1/html5/thumbnails/8.jpg)
Collision distance less than molecular diameter
Molecular Proximity = Big Issue
![Page 9: Automated Protein Dilutor and Interferometer](https://reader035.fdocuments.us/reader035/viewer/2022062813/56816696550346895dda7dd1/html5/thumbnails/9.jpg)
Energy exchanges Ionic strength effects pH dependence Predict protein
behavior
Bimolecular Collisions
![Page 10: Automated Protein Dilutor and Interferometer](https://reader035.fdocuments.us/reader035/viewer/2022062813/56816696550346895dda7dd1/html5/thumbnails/10.jpg)
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
Where does dilution take place in drug development process?
![Page 11: Automated Protein Dilutor and Interferometer](https://reader035.fdocuments.us/reader035/viewer/2022062813/56816696550346895dda7dd1/html5/thumbnails/11.jpg)
1. Remove aliquot into waste container 2. Add aliquot of buffer solution3. Stir media for 20 seconds 4. Light scattering - Wyatt Dawn HELEOS II5. Find activity coefficient with interferometer 6. Process repeats fifty times in 12 hours
Dilution Sequence
![Page 12: Automated Protein Dilutor and Interferometer](https://reader035.fdocuments.us/reader035/viewer/2022062813/56816696550346895dda7dd1/html5/thumbnails/12.jpg)
Dilutor Diagram
![Page 13: Automated Protein Dilutor and Interferometer](https://reader035.fdocuments.us/reader035/viewer/2022062813/56816696550346895dda7dd1/html5/thumbnails/13.jpg)
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
Interferometer
![Page 14: Automated Protein Dilutor and Interferometer](https://reader035.fdocuments.us/reader035/viewer/2022062813/56816696550346895dda7dd1/html5/thumbnails/14.jpg)
Interferometer Diagrams
![Page 15: Automated Protein Dilutor and Interferometer](https://reader035.fdocuments.us/reader035/viewer/2022062813/56816696550346895dda7dd1/html5/thumbnails/15.jpg)
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
Interferometer Processing
![Page 16: Automated Protein Dilutor and Interferometer](https://reader035.fdocuments.us/reader035/viewer/2022062813/56816696550346895dda7dd1/html5/thumbnails/16.jpg)
1. Program Hamilton dual syringe dilutor2. Hamilton to Spectrocell communication3. Retrofit Mini-Motor to drive PTFE mixing shaft 4. Secure mixing motor to cell cuvette5. Concentration calculation program6. Interferometer – concentration measurement7. Solid-state camera to pixels, TV 8. Algorithm to process fringe displacement
Objectives
![Page 17: Automated Protein Dilutor and Interferometer](https://reader035.fdocuments.us/reader035/viewer/2022062813/56816696550346895dda7dd1/html5/thumbnails/17.jpg)
Enable Motor Controller Communication from Syringe Dilutor
![Page 18: Automated Protein Dilutor and Interferometer](https://reader035.fdocuments.us/reader035/viewer/2022062813/56816696550346895dda7dd1/html5/thumbnails/18.jpg)
Item Vendor Cost
Dell Laptop UNH Computer Store $429.00
Laptop set-up cost UNH Computer Store $25.00
Stirring System Spectrocell, Inc. $614.00 + $34.00 shipping
*ML541C syringe dilutor Hamilton $3949.70
Total: $5051.70
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
![Page 19: Automated Protein Dilutor and Interferometer](https://reader035.fdocuments.us/reader035/viewer/2022062813/56816696550346895dda7dd1/html5/thumbnails/19.jpg)
Timeline
![Page 20: Automated Protein Dilutor and Interferometer](https://reader035.fdocuments.us/reader035/viewer/2022062813/56816696550346895dda7dd1/html5/thumbnails/20.jpg)
Diagrams
![Page 21: Automated Protein Dilutor and Interferometer](https://reader035.fdocuments.us/reader035/viewer/2022062813/56816696550346895dda7dd1/html5/thumbnails/21.jpg)
Diagrams (continued)
![Page 22: Automated Protein Dilutor and Interferometer](https://reader035.fdocuments.us/reader035/viewer/2022062813/56816696550346895dda7dd1/html5/thumbnails/22.jpg)
Q & A