DNA molecule
deciphered by
Watson & Crick
Enzyme
involved in the
synthesis of
nucleic acid
isolated
1950s
Harris &
Watkins fuse
mouse and
human cells
Automatic
protein
sequencer is
perfected
1960s
First generic
markers for
human disease
are found
Human Genome
Project
Recombinant
human insulin
Nearly 5 million
acres of biotech
crops grown
Commercial
biotech
companies
formed
Development of
leading
biotech
products
1970s
1980s
1990sToday
Stem cell
applications
Gene therapy
Probiotics
Efficient
bioconversion
of cellulose
into fuel
Tomorrow
Proteins
(including therapeutic antibodies)
MW: 180 Da
Acetylsalicylic Acid
(Aspirin)
Organic Chemistry
Biochemistry
Recombinant DNA Technology
Design the
Process
Design the
Plant
The Molecule
Challenges and Trends in Measurement
CH1
S
S
VL
S
S
S
S
N-terminal glutamine
cyclization to pyregluteic acid
S
S
CL
Disulphide scrambling
misfolds
CH2
CH3
S S
S S
N-linked glycoforms
S S
S S
Elimination of subjectivity
Visual inspection- presence of particles, color
Move toward automated instrument-based inspection
1990
- SDS-PAGE
- ELISA
- Chromatography
- Some MS
2020
MS based analysis:
- Host Protein Fingerprint
- Top Down Analysis
- Routine analysis of CQA
1990
100
1993
10
1997
2000
0.1
2003
0.01
2005
0.001
2008
0.0001
Dissociable Dimer
Subvisible Particles
Laser Diffraction*
DLS*
AUC
Microscope
SEC
nm
10
Visual Inspection
L.O. (HIAC)
100
monomers &
oligomers
10
100
subvisible particles
mm
cm (107 nm)
visible particles
* qualitative
Potency measurements
In Vitro Binding Assays
Target
SA biotin FcRn
Target
Target
Pro Luciferase
Developing the
Manufacturing Process
Mammalian Cells:
Core Manufacturing Technology
MOLECULAR CELL BIOLOGY 2ed by Lodish et al. (c) 1990 by Scientific American Books.
Used with permission by W.H. Freeman and Company. Illustration by Tomo Narashioma
Poolofstable
transfected cells
Individualencapsulation
intogelmicrobead
Captureandstainingof
secretedproducts
FACSquantitation
andsinglecell
sortingofhigh
producingsubclone
96wellplate
Day9Imaging
1020Clones
ScaleUp
24Clones
(HTRF)
T25
T75
25mLshake
flasks
24wellplate
Poorgrowers
eliminated
11DayFedBatchAssay
Gene
transcription
metabolic
engineering
ENVIRONMENTAL
DESIGN
bioreactor conditions
De-bottleneck
expression pathway
CELLS
MEDIA DESIGN
nutrients & metabolites
5 incubators
Temp. CO2, O2. Humidity, Mixing
1260 mini-reactors
8 sources for liquid addition ( 1.5ul)
pH control (< 0.1)
OD measurement
Integrated database
Link to other systems
TECAN, Guava etc.
Filterplates with
96 conditions
Batch-binding
Stack Filterplate
on ELISA plate
Centrifuge
/ Vacuum filter
Wells: 800 L
Resin: 100 L
Liquid: 365 L
Collect 300 L
Supernatant
Plant
Design
Process
Design
Process
Control
Quality/Safety
PLANT
Raw
Materials
PROCESS
Environment
Process Design
Productivity and Quality
Product
TMP, flow,
conductivity
UV
pH,
conductivity
C, time,
concn
Water
Drying time,
temperature
pH,
conductivity
UV, flow, TA
Raw materials
Process aids
On-line HPLC,
pH, OD, temp,
NIR, MS, CZE
pO2, pCO2
Viral detection
Product titre
metabolites
Viability,
Product variants
metabolites
HPLC/CZE
TMP,
Aggregates
conductivity
variants,
UV
contaminants
pH,
conductivity
HPLC/CE
C, time
Water
Environmental
Drying time,
NIR, Moisture
temperature
content
pH,
conductivity
HPLC/CZE/
UV,
MS
Biosensor
Plant Design
Capital Cost
engineering construction
materials
Operating Cost
utilities
maintenance
environmental control/monitoring
2.
REMOVABLE
PANEL
UTILITY
HEADER
Degree of Integration
Comprehensive
Prefab
integrated prefab structure /
facility / process
Box-in-a-Box
integrated pre-fab facility
within a stick-built structure
Partial Prefab
prefab assemblies for insertion
in stick-built structure and
facility
Resolves mixtures of
organisms
Quantitative
Sub-species
resolution (strain
genotyping)
Days
80
Confirm contaminant
ID contaminant
60
60
40
35
20
0
10
11
With Rapid
Methodology*
Without Rapid
Methodology**
With Universal
Pathogen Detector
1H
Riboflavin
1H
peaks.
20
water
D M SO
11
4
14
17,
26a
OH
OH
OH
OH
26b
22
24
18
44
T M S,
internal std
21
Riboflavin
1H NMR spectral comparison between riboflavin lot # 0010011629
and USP reference standard lot# N0C021
Riboflavin lot# 0010011629
Acknowledgments
IzydorApostol Analytical&FormulationSciences
ThomasArroll CellularResources
Pavel Bondarenko Formulation&AnalyticalResources
Houman Dehghani CellularResources
GregFlynn Analytical&FormulationSciences
AndyGoetze Analytical&FormulationSciences
DrewKelner Analytical&FormulationSciences
BrentKendrick AnalyticalSciences
DuncanLow ProcessDevelopment
JeffMcGrew CellSciences&Technology
LindaNarhi Formulation&AnalyticalResources
TimothyOsslund Analytical&FormulationSciences
SureshVunuum PurificationProcessDevelopment
Siowfong Wee Analytical&FormulationSciences
Zhongqi Zhang Formulation&AnalyticalResources