Dedicated Analytical Solutions Near-Infrared Spectroscopy as a Process Analytical Technology in the Biotech Industry Presented to the Advisory Committee for Pharmaceutical Science Food and Drug Administration April 13, 2004 By Robert A. Mattes Foss
Jan 17, 2018
Dedicated Analytical Solutions
Near-Infrared Spectroscopy as a Process Analytical Technology
in the Biotech IndustryPresented to the
Advisory Committee for Pharmaceutical Science
Food and Drug AdministrationApril 13, 2004
ByRobert A. Mattes
Foss
Dedicated Analytical Solutions
Near-infrared Spectroscopy Spectral Region
UV VisibleNear-Infrared
Mid-Infrared
25004,000
Functional Group Analysis
Constituent AssayAppearance
Elemental
70014,285
40025,000
200 50,000
25,000 nm400cm-1
Dedicated Analytical Solutions
Pharmaceutical ManufacturingNIR Applications
Incoming Raw
Materials
Material Dispensary
In-Process: Fermentation
and Cell Culture Monitoring Warehouse
Shipping
Lyophilization
Dedicated Analytical Solutions
BioPharm Process Reactor
Control feedback
On-line monitoring of critical process
parameters
Process control
Raw MaterialsProcess output
Closed loop control (Real-Time)
TemperatureSparging
NIR monitoringNutrient levels
Control Capabilities with NIR/PAT
Moisture level
Melt detection
In Lyo
Process Improvement
Dedicated Analytical Solutions
Organism Supplier Product Fluid Behavior
Analyte (s)
Escherichia coli (unicellular)
Eli Lilly BST (bovine somatotropin)
Non-viscous Newtonian
biomass, substrates, acetate (by-product), product
Streptomyces fradiae
(filamentous)
Eli Lilly Tylosin Viscous non-Newtonian
biomass, substrates, product
Pichia pastoris (unicellular)
British Biotechnology,
PLC
Human Protein
Viscous Newtonian
biomass, substrates, product
CHO-K1 (animal cell-line)
British Biotechnology,
PLC.
Human Protein
Non-viscous Newtonian
Biomass, substrates, product
Industrial Organisms Studied NIR Bioprocess Method Development
Dedicated Analytical Solutions
NIR Spectral Acquisition and Analysis for Bioprocesses
0
1
2
3
400 700 1000 1300 1600 1900 2200 2500Wavelength (nm)
Log(
1/T)
Dedicated Analytical Solutions
NIR Spectral Analysis Analytical Region for Key analytes
0
1 6 00
0
1600 1625 1650 1675 1700 1725 1750Wavelength (nm)
Seco
nd-d
eriv
ativ
e of
Log
(1/T
)
Glycerol, acetate, biomass
t=0, t=27 hr
Dedicated Analytical Solutions
NIR Bioprocess Trends Chart
0
1
2
3
4
0 2 4 6 8 10Time (hours)
Gly
cero
l (g/
L)
0
1
2
3
4
Bio
mas
s (g
/L)
Dedicated Analytical Solutions
NIR Bioprocess Analytical ResultsAnalyte Strategy Ref Error Model SEC/SEP
Biomass (g/L) At-Line 0.7 g/L PLS 1.2/1.4
Acetate (g/L) At-Line 0.7 g/L MLR 1.2/0.7
Glycerol (g/L) At-Line 0.7 g/L MLR 1.1/0.7
Ammonium (mmol/L) At-Line 8 mmol/L PLS 13/7
Carbon Nutrient (%) In-Line 0.05% MLR 0.06/0.06
Glycerol (g/L) Lab 0.3 g/L MLR 0.3/0.3
Biomass Lab 0.2-0.4 g/L MLR 0.2/0.2
Dedicated Analytical Solutions
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).
Dedicated Analytical Solutions
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 <1
minute Monitoring and closed-loop
control Pilot plant/scale-up operations
and manufacturing
Direct Sample Interface
Dedicated Analytical Solutions
Summary of NIR Results for CHO cell work
Analyte Range SEC R2 SEP
Ammonia 0.5-2.0mM
0.032 0.99 0.036
Glucose 0.7-3.5g/L
0.07 0.99 0.07
Lactate 0.3-1.2g/L
0.008 0.99 0.014
Glutamine 2-6mM 0.12 0.99 0.31
Multifactor PLS calibrations developed on second derivative spectra with 217 samples
Dedicated Analytical Solutions
Enlarged View of Second Derivative Spectra
Used for Analysis of Lyophilized Product
-0.10
-0.08
-0.06
-0.04
-0.02
0.00
0.02
0.04
0.06
0.08
0.10
1800 1850 1900 1950 2000 2050 2100 2150 2200
Wavelength (nm)
2nd D
eriv
ativ
e Sp
ectra
Dedicated Analytical Solutions
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
NIR Benefits for the PAT Biotech Initiative