NearInfrared Spectroscopy as a Process Analytical Technology in the Biotech Industry

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Near-Infrared Spectroscopy as a Process
Analytical Technologyin the Biotech Industry
Presented to the Advisory Committee for
Pharmaceutical Science Food and Drug
Administration April 13, 2004 By Robert A.
Mattes Foss
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Near-infrared Spectroscopy Spectral Region
Near-Infrared
UV
Visible
Mid-Infrared
200
700
400
2500
25,000 nm
14,285
25,000
50,000
4,000
400cm-1
Functional Group Analysis
Constituent Assay
Elemental
Appearance
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Pharmaceutical ManufacturingNIR Applications
Incoming Raw Materials
Material Dispensary
Lyophilization
In-Process Fermentation and Cell Culture
Monitoring
Warehouse Shipping
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Control Capabilities with NIR/PAT
Control feedback On-line monitoring of critical
process parameters
Process control
Process Improvement
Closed loop control (Real-Time)
BioPharm Process Reactor
Temperature Sparging NIR monitoring Nutrient
levels
Moisture level Melt detection In Lyo
Process output
Raw Materials
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Industrial Organisms Studied NIR Bioprocess
Method Development
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NIR Spectral Acquisition and Analysis for
Bioprocesses
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NIR Spectral Analysis Analytical Region for Key
analytes
t0, t27 hr
Second-derivative of Log(1/T)
Glycerol, acetate, biomass
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NIR Bioprocess Trends Chart
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NIR Bioprocess Analytical Results
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Near IR applied to Mammalian Cell Cultivation

• Industrial CHO-K1 cell line
• In-situ NIR using transflectance measurements
  (1.2 mm pathlength)
• 2 Liter bioreactor run for 168 240 hours,
  including a temperature change
• Developed NIR calibrations for
• Glucose
• Glutamine
• Lactate
• Ammonia

Ref SA Arnold, J Crowley, N Woods, LM Harvey,
and B McNeil, Biotech and Bioeng 84(1), 13-19
(2003).
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Direct In-Line Analysis in Mammalian Cell
Cultivation

• Direct fermenter interface
• Standard Ingold ports
• Optimized for specific analytes/matrices
• Dedicated analysis on defined media
• Simultaneous results lt1 minute
• Monitoring and closed-loop control
• Pilot plant/scale-up operations and manufacturing

Direct Sample Interface
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Summary of NIR Results for CHO cell work
Multifactor PLS calibrations developed on second
derivative spectra with 217 samples
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Enlarged View of Second Derivative Spectra Used
for Analysis of Lyophilized Product
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NIR Benefits for the PAT Biotech Initiative

• NIR offers the advantage of real-time analysis in
  biotech sterile environments and the possibility
  of closed-loop feed back process control
• Benefits in process
• Non-invasive analysis
• Process optimization
• Waste reduction
• Better understanding of processes and
• Safer, improved, more consistent product