THE IMPACT OF MICROANALYTICAL INSTRUMENTATION ON PAT APPLICATIONS

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THE IMPACT OF MICRO-ANALYTICAL INSTRUMENTATION
ON PAT APPLICATIONS
M Koch
Center for Process Analytical Chemistry (CPAC)
University of Washington
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PAT What is it?

• Applications of
• Process analytical chemistry tools
• Process control strategies
• Information management tools
• Data analysis tools
• GOAL - PROCESS UNDERSTANDING

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ITS ORIGIN IS IN TRADITIONAL ANALYTICAL SCIENCES

• SPECTROSCOPY
• CHROMATOGRAPHY
• ELEMENTAL ANALYSIS
• MICROSCOPY
• THERMAL
• POLYMER CHARACTERIZATION
• MADE PORTABLE FOR PROCESS ANALYSIS (real time
  analysis)

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Real-time Data Acquisition . . .

• . . . has provided researchers with a video of
  a process.
• Deliverables
• Composition
• Reaction Pathways Kinetics
• Emission Monitoring
• Hazard Evaluation Issues
• On-line Feasibility Studies
• PROCESS UNDERSTANDING

Courtesy Dow Chemical
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What's Appropriate for PAT?

• Anything ...
• That makes a measurement that cant presently be
  made...
• Thats cheaper and more reliable
• Where more (measurement points) results in better
  control...
• Depart from traditional analytical sciences
  techniques if appropriate

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Tasks for Instrument Developers

• Create fully integrated analyzer systems
• Develop measurement approaches that are
• more information rich (inferential)
• Revisit underutilized techniques
• (e.g.. optical coherence, Raman, dielectrometry,
  surface plasmon resonance)

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Relevance ofOptical Low Coherence Reflectometry
(OLCR)

Lloyd Burgess, UW Chemistry Dept
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Optical Low Coherence Reflectometry

• Profile Depends On
• Sample Thickness
• Particle Size
• Particle Concentration
• Particle Shape
• Morphological Domains
• Change in RI
• Probe Geometry
• Source Coherence Length

• Used to Monitor
• Coke deposition on spent catalyst
• Coating thickness
• Membrane micropore analysis
• Attrition milling processes
• Polymer thickness in drug delivery patches
• Paint curing times
• E. coli and yeast at high concentrations

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Multiple Layer Polymer Materials
Scattering materials
Drug delivery patch
Distance in micrometers
Distance in micrometers
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Measurement of particle size and concentration
Polystyrene microspheres in deionized water
19nm 90nm 1
0.25 - 10 308nm
log10 ( I / I o)
log10 ( r R3)
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Evaluating Raman Spectroscopy for Process
Monitoring and Materials Characterization

• Brian J. Marquardt
• Dave Veltkamp, and Brian Dable
• Center for Process Analytical Chemistry
• University of Washington

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Advantages of Raman Spectroscopy

• Little or no sample preparation is required
• Water is a weak scatterer - no special
  accessories are needed for measuring
• aqueous solutions
• Fiber optics (up to 100's of meters in length)
  can be used for remote analyses
• Raman bands can be easily related to
• chemical structure (very good for
  fingerprinting)
• Ideal for both organic and inorganic species

Brian Marquardt UW CPAC
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Raman Sampling Applications
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Parameter Detection with Fringing Electric Field
Dielectrometry Sensors

• Alexander Mamishev, UW Electrical Engineering

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Sensing Possibilities

• Fringing electric fields can detect various
  characteristics of a sample.

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• Experimental Results
• Paper Pulp
• Pharmaceutical Products
• Food Products
• Composite Materials
• Plastics

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APPLICATIONS OF SURFACE PLASMON RESONANCE SENSORS

Electrical Engineering Department, Departments
of Genetics and Medicine, University of
Washington
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Target Applications for Real-Time Biosensors

• High throughput drug screening
• Automated protein purification
• Detection of toxins, pesticides, and food poisons
• Bioagent identification

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Protein purification system
Pump 1
Column 1
Column 2
Pump 2
Filter 1
Sensor 1
Fermentor
Sensor 2
Pump 3
Filter 2
Product Out
Waste
Pump 4
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Sensors in Bioprocess Control Harry
Lam Department of Manufacturing
Sciences Genentech, Inc
Genentech, Inc.
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The Need for BioProcess Control

• To maintain consistent process performance
  (productivity, quality) throughout the
  development cycle (RD to Manufacturing).

Genentech, Inc.
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Sensors for Bioprocess Control

• Measurements for process or system analysis
• Search for underlying functional relationships
• In depth analysis of the interaction of the
  organisms with their environment
• Provide capabilities for process control
• Setting up and maintaining the optimum
  environmental conditions for growth and/or
  formation of product

Genentech, Inc.
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Measurements

• Biological
• Chemical
• Physical

Genentech, Inc.
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Biological Measurements

• Cell Density
• Cell Viability
• Cell Size
• Morphology
• Cellular assays
• Molecular/Genetic Assays

Genentech, Inc.
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Chemical Measurements

• Media
• Carbohydrates - e.g. glucose, galactose
• Complex medium protein hydrolysates, yeast
  extracts, etc.
• Amino acids
• Salts
• Lipids - Linoleic Acid
• Hormones, growth factors (serum, insulin)
• Vitamins
• Trace elements - e.g. metals (Fe, Mn, etc.)
• Antifoam
• F-68
• Antibiotics
• Methotrexate

Genentech, Inc.
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Chemical Measurements (cont.)

• Product
• Concentration
• Quality
• By-products
• Organic acids acetate, lactate, etc
• Proteins
• Ammonia
• Chemical environment
• pH
• Dissolved O2, CO2 (pO2, pCO2)
• Osmolality
• Off-gas
• O2 (OUR)
• CO2 (CER)

Genentech, Inc.
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Physical Measurements

• Temperature
• Agitation
• Pressure
• Level (volume)
• Weight
• Broth density
• Viscosity

Genentech, Inc.
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What can be routinely measured on-line today in
cell culture systems?

• Cell density (OD)
• Temperature
• Agitation
• Pressure
• Culture Volume
• pH
• Dissolved O2
• Off-gas composition

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Requirements for Process Sensors

• Fully cleanable and sterilizable
• Long-term stability and accuracy
• Fast response
• No flow dependence
• No interference
• Air bubbles (O2 and CO2) or by microbes
• Complex media
• No fouling
• Low maintenance
• Small size

Genentech, Inc.
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REVOLUTIONARY ADVANCES THAT ARE AFFECTING
CHEMICAL MEASUREMENTS

• MINIATURIZATION
• ENGINEERED MATERIALS
• ELECTRO-OPTICS
• MEASUREMENT INFORMATION PROCESSING

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CPAC is a Consortium of Industrial Sponsors,
National Laboratories and Government Agencies
? Established in 1984, the Objective of the
Center is to Address Challenges in Real-Time
Analysis and Process Control
? Provides a Forum for Multidisciplinary
Participation in Process Analytical Technology
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Multidisciplinary
? Chemistry ? Chemical Engineering ? Electrical
Engineering ? Bioengineering ? Forestry ?
Mechanical Engineering ? Food Science
Engineering ? Health Sciences
Presently supporting 20 projects at 5 Universities
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CPAC INITIATIVES

• OPEN
• NEW SAMPLING AND SENSOR INITIATIVE (NeSSI)
• CHEMOMETRICS ON-LINE INITIATIVE (COPA))
• CLOSED
• MICRO-INSTRUMENTATION FOR HIGH THROUGHPUT
  EXPERIMENTATION PLATFORM
• FERMENTATION MONITORING AND CONTROL PLATFORM
  (FermI)

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Improving Process Optimization using
Miniaturization Technology

• MICRO-INSTRUMENTATION AND MICRO-REACTORS

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SOME RELEVANT CPAC RESEARCH EFFORTS IN
MINI-INSTRUMENTATION

• MICRO-LC
• SURFACE TENSION DETECTION
• GCxGC
• FRINGE FIELD SENSORS
• VAPOCHROMIC SENSORS
• RAMAN
• GRATED LIGHT REFLECTIVE SPECTROSCOPY
• REFLECTOMETRY
• FLOW-THROUGH PARTICLE ANALYZER
• SURFACE PLASMON RESONANCE
• MINI-MASS SPECTROMETER
• MINI-NMR
• ULTRASOUND

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Process Liquid Chromatography and
Microfabricated Liquid Analyzers
Colin D. Costin, Adam D. McBrady, Ana K.
Torgerson, Robert E. Synovec Department of
Chemistry, University of Washington Collaboratin
g Personnel Dr. Milton E. McDonnell
(Honeywell)
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?-Refractive Index Gradient(?-RIG) Detection
Mechanism
Mobile Phase Inlet
Sample Inlet
Position Sensitive Detector
?
Deflected Beam Is Measured

Laser Beam
To Outlet
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Application as a Micro-scale Molecular Mass
Sensor (?-MMS)
Outlet
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m-MMS, Measuring m-RIG Signal at Two
PositionsReal-Time, Dual-Beam Detection
Configuration
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Typical Molecular Mass Calibration for Linear PEGs
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Process Monitoring with the ?-MMSCan readily
monitor peptide or protein synthesis

• Amino Acid and Peptide solutions for monitoring
  protein synthesis or digestion
• Quickly able to detect addition or subtraction of
  amino acids

Gly-Gly-Tyr-Arg
Gly-Gly-His
Gly-Gln
Gly
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Sandia microChemLab Modules
Preconcentrator/ Thermal Desorber
4 SAW Array
One Meter GC Column
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A fully autonomous hand-held micro analyzer

Containing Multiple coated/packed GC columns
w/multiple coated SAW detectors
and Multiple LC columns with fluorescence
electrochemical detection
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Synovec CPAC UW
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The ILD-50 Industrial Mass Detector
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Development of a Micro NMR System

• UCD NMR Facility
• Micro Instruments and Systems Laboratory
• Department of Electrical and Computer Engineering
• Department of Biological and Agricultural
  Engineering
• University of California, Davis

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Initial Results
NMR spectrum of a 3 micro liter water sample
using a RF micro coil
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ADVANCES IN SENSORS AND CONTROLSHIGHLIGHT THE
NEED FOR IMPROVED SAMPLING TECHNOLOGIES
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(New Sampling/Sensor Initiative) NeSSI
ExxonMobil, Dow, CPAC
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Todays Typical Production Sampling Technology
Courtesy of a major petrochemical facility
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What is NeSSI?
Picture courtesy Swagelok
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Evolution of NeSSI
Micro Analytical (enabled by Technology Advances)
End User Value
Gen III Circa 200?
Smart Devices (enabled by Communication Standard)
Gen II Circa 2005
We are Here
Miniature/Modular (enabled by Mechanical
Standard)
Gen I Now
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NeSSI Sandwich concept
Standard Connectivity Interface
Ethernet LAN
DevelopmentFocus can beon Sensors !
CANbus
Analyzer Controller
SAM
Standard Sampling Interface
Substrate
PDA
R. Dubois, Dow
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The NeSSI Platform Will Also Become the Base for
a Micro-Analytical Lab
NMR MS NIR RAMAN LC GC SIA Particle
Size Viscosity Moisture Conductivity Mass
Flow Etc.
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Raman Sensor Module
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Miniature Surface Plasmon Resonance (SPR) Sensor
Platform
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mSequential Injection-Lab On Valve System
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NeSSI with an Array of Micro-Analytical
Techniques will Impact

• Process Control
• Process Optimization

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Fermentation Platform Initiative (FermI)
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Goals of FermI

• Application and analysis of current fermentation
  tools
• Adaptation and use of known analytical techniques
  to fermentation problems
• What other existing tools might be useful?
• Development and application of new fermentation
  analytics
• What can emerging tools offer?
• Provide training and experience in fermentation
  operations and issues.
• What new skills and considerations are important?
  

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CPAC Fermentation Platform

• Instrumentation
• pH, DO, Temp., agitation
• NIR reflectance
• Raman BallProbe
• Visible (Optical Density).
• Dielectric spectroscopy
• Mass spectroscopy
• Reflectrometry
• Grated Light Reflective Spectroscopy
• Surface Tension
• Micro-LC, SIA
• Evaluate Sampling Technologies

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Fermentation Sampling Considerations

• Representative sample
• Sample size
• Sterility requirements
• Utilities considerations
• Disposal considerations
• Liquid versus vapor samples
• Sample preservation

Genentech, Inc.
63
Sterile Sampling Designs

• Sample location
• Sample withdrawal position
• Method of connection
• fluid velocity profile
• Containment considerations
• Sampler design
• Materials of construction
• Process and sample variables

Genentech, Inc.
64
Future Work

• Continue with scoping activities to evaluate
  analytical monitoring tools
• Implement and evaluate NeSSI systems as sampling,
  sensor, and process control interfaces
  (broth and vapor)
• Begin to look at chemometrics tools to model
  batch variability and data fusion approaches

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Chemometric Analysis of Production Data

• Goal
• To develop automated tools to evaluate
  production data.
• Implementation of chemometrics to quantify
  process performance.
• Application of Principal Component Analysis (PCA)
  to perform automated pattern recognition.

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IT IS GOING TO BE AN EXCITING TIME
IMPLEMENTING A NEW TECHNOLOGY INFRASTRUCTURE IN
PHARMACEUTICAL AND BIOLOGICAL MANUFACTURING