GMp

1. Merck KGaA
Darmstadt, Germany
Akshat Gupta and Elizabeth Goodrich
October 10, 2019
An Efficient and cGMP-
Friendly Solution to
Diafiltration for
Intensified or Continuous
BioProcessing

2. The life science business of
Merck KGaA, Darmstadt, Germany
operates as MilliporeSigma
in the U.S. and Canada.

3. Agenda
01 The Case for Continuous Diafiltration
02 An Efficient Solution to Continuous Diafiltration
03 24-Hour Process Demonstration
04 Conclusions

4. The Case for
Continuous
Diafiltration
01

5.  Deliver target molecule in final formulation buffer and at specified concentration
 Removes low MW solutes
 Typical mass loadings: 150 g mAb/m2-hr or ~ 0.5 kg/m2 for typical 3-hr processing time
 Largest systems of 6 high holder with 120m2 limited to 60 kg batches
 Large tank & footprint, high capital and long lead times
 High working volume & potential product recovery dilution
 Long setup (install, flush, IT) and turnaround (flush, clean, NWP, sanitize, store)
General mAb process:
Bioreactor Clarification Protein A
Capture
Cation
Exchange
Purification
Anion
Exchange
Polishing
Viral
Inactivation
Virus
Filtration
Concentration
Diafiltration
Formulation
Sterile Filtration
Final Fill
An Efficient and cGMP-Friendly Solution to Diafiltration for Intensified or Continuous BioProcessing | 10 October 2019
Background: Batch Formulation Step
5

6. Evolution to Continuous Processing
Intensified Upstream
& harvest
Intensified
Capture
In-Line VI
In-Line Buffer
Flow Through
Polishing
SPTFF &
Continuous
DF
• Manufacturers pursuing continuous processes are starting to address this step
• Existing solutions present implementation challenges
• Can we do better than batch UF/DF?
An Efficient and cGMP-Friendly Solution to Diafiltration for Intensified or Continuous BioProcessing | 10 October 2019
6

7.  Typically comprised of 3 steps (UF, DF, UF) plus Product Recovery
 Diafiltration occurs at constant volume and constant (specified) concentration
DF consumes the majority of process time (membrane area) of the overall operation
Final Formulation Batch UF/DF Operation
An Efficient and cGMP-Friendly Solution to Diafiltration for Intensified or Continuous BioProcessing | 10 October 2019
7
Initial concentration
Diafiltration
Final concentration
0
20
40
60
80
100
120
140
160
0% 20% 40% 60% 80% 100%
g/L
concentration
process time
UF UF
DF
~10 to 70g/L ~8 to 10 Diavolumes @
Copt (70g/L)
~70 to
150g/L

8. Standard batch DF optimizes membrane area and buffer usage
• Diafiltration at constant volume (i.e level control) results in
more efficient buffer exchange versus a series of UF
concentrations followed by buffer dilution
• Diafiltration at the optimal protein concentration provides the
best balance between flux during DF and volume needed to
be diafiltered
• Results in lowest required membrane area
An Efficient and cGMP-Friendly Solution to Diafiltration for Intensified or Continuous BioProcessing | 10 October 2019
8
DF Optimization Parameter = CbDF * JDF
Optimum Cb
for DF