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

Project Introduction Definition Biotech Overview Objectives Budget Timeline Diagrams

Agenda

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

Design and construct a bench-top automated protein dilutor and interferometric device to

measure and monitor concentration.

Project Definition

Biotech Overview

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

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

Collision distance less than molecular diameter

Molecular Proximity = Big Issue

Energy exchanges Ionic strength effects pH dependence Predict protein

behavior

Bimolecular Collisions

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?

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

Dilutor Diagram

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

Interferometer Diagrams

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

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

Enable Motor Controller Communication from Syringe Dilutor

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

Timeline

Diagrams

Diagrams (continued)

Q & A