Ultrafiltration/Diafiltration Process

Development of High Concentration/

Viscosity Applications
Frederic Sengler, Josselyn Haas, MilliporeSigma, Molsheim, France, Renato Lorenzi, MilliporeSigma, Milan, Italy

Screen Selection The mass transfer coefficient and Cw was determined by calculating
absolute slope of permeate flux versus bulk concentration as
Introduction We currently offer four different screens: (A, C, D and V) in the Pellicon® described by the stagnant film model equation taking into account
data range before pump turn down:
cassette format range.
Formulating a highly concentrated product
A screen: A tight screen that is used for dilute protein solution
that is suitable for subcutaneous delivery is
or low viscosity solutions. The tight weave is not suitable for highly
key to achieving injectable volumes, typically
concentrated protein solutions as the pressure drop is unacceptably high
around 1 mL. at greater viscosities. However, this screen typically provides superior
Newly designed TFF cassettes capable of flux performance.
processing high viscosity product streams Where
C screen: A coarse screen that is used for product streams with
were tested using human IgG (Seracare) as J = normalized permeate flux
viscosity up to 15 cP. The C screen provides good flux performance
Cw = protein concentration at the membrane
a model feed. The results showed that the but the pressure drop is above the limit of usage when the viscosity
Cb = protein concentration in the bulk
pressure drop along the feed channel was increases.
Cp = Protein concentration in the permeate (considered = 0)
reduced by more than 50% compared with V screen: A suspended C screen. The V screen gives lower pressure k = mass transfer coefficient
regular TFF devices while only sacrificing 15% drops than the C screen, but the flux performance is much lower
of the flux and the mass transfer coefficient. Initial and final protein concentration was determined by UV
compared to C screen.
Compared with more open channel TFF measurement.
D screen: This new A Screen
devices with a suspended feed screen, the 0.4 mm thick
screen has a coarser
new high viscosity TFF cassettes can reach mesh and altered Experimental results are summarized in the following table,
C Screen
the same final concentration with nearly weave compared to the 0.53 mm thick comparing D versus C screen performances:
double the mass transfer coefficient over the standard C screen. It
course of the process. was designed to offer V Screen
lower pressure drop
0.93 mm thick 3 LMM 5 LMM 7 LMM
In this poster, we focus on the various
screens that are available in the Pellicon® without the large flux
performance penalty D Screen Screen D C D C D C
cassette family, especially the D screen 0.61 mm thick
found when using a
which was developed to achieve formulations suspended screen k (LMH) 13,21 20,17 33,73 34,4 35,33 36,83
with a viscosity higher than 15 cP within an technology Figure 3: Pellicon Cassette Screens ®

acceptable pressure drop and with a high

Cgel (g/L) 236 230 230 209 229 217
flux. A robust, scalable comparison trial
between the different screens was performed
Max g/L 254 221 275 231 275 226
in order to compare these screens.
Comparison Study of C and
D Screen at High Protein Table 1. Process concentration summary: D and C
screen performances comparison
Concentrations
As the viscosity increases, so does pressure drop with a
proportional relationship. It has been verified that in a well An internal study was performed in order to evaluate screened module
The D screen flux along the concentration step is similar to the C
designed TFF UF system the component that has the greatest resistance (or pressure drop or DP) vs. protein (IgG) concentration.
screen one at feed flow higher than 5 L/min/m², but D screen was
effect on overall system resistance (pressure drop) is the TFF The pressure drop across the feed channel for cassettes with different able to reach a 20% higher final concentration.
cassette itself. (Figure 1). screens is shown in Figure 4. At a given feed flow, the pressure drops
D screen average flux is 75% compared to C screen at 3L/min/m²,
increase with the “tightness” of the screen. Since the pressure drop
but still a 10% higher final concentration was reached.
35 (dark blue and light blue lines) is related to the protein concentration
holder
and its viscosity (green line), the lower the feed channel resistance the In all the experiments, D screen was able to reach a higher final
30 90% elbow
concentration than C screen.
higher the protein concentration that can be achieved.
straight pipe
25 tank return
block valve
20 Module resistance and IgG viscosity as a function of
Figure 4:
psid

retentate valve concentration at 25°C - Buffer: Milli-Q® ultrapure water

15 cassette 50
45 Viscosity
90
80
D screen and C
screen cassette
Conclusion
D screen
10 40 High protein concentration by TFF UF is a challenging
C screen 70 resistance
Resistance (psi/LMM)

35 application. From an operational point of view, the increase

5 60 vs. IgG
Viscosity (cP)

30
50 of viscosity and module resistance with the increase
25 concentration of protein concentration obliges the operator to find a
0 40
5 cp 10 cp 15 cp 20 cp 20 (internal data). compromise between pressure drop (module resistance)
30
15 and flux at final concentration.
20
Figure 1: TFF System components pressure drop 10
5 10 A variety of TFF cassette designs exist and the choice of
0 0 the most appropriate TFF cassette is key for an efficient
0 50 100 150 200 250 300 and high performance process.
The pressure drop of ultrafiltration cassettes (module IgG Concentration (mg/mL)
resistance) due to the increased viscosity of the protein The new Pellicon® 3 cassettes with D screen, when
solution during concentration, is the most important factor accompanied by the best working procedure, are able
affecting the ability to achieve high final product concentrations The new D screen cassette has a pressure drop about 50% of the to provide higher final product concentration with similar
in TFF UF applications for protein. C screen pressure drop. A comparative concentration step was also flux performance of a tighter screened cassette. This
run at 3 different feed flows (3 – 5 and 7 L/min/m²) post flux vs. performance similarity is dependent on being able to
Different module designs exhibit different pressure drops; provide a reasonable feed flow rate.
Figure 2 demonstrates how utilizing a conventional screen TMP excursion with a 20 g/L solution of human IgG to determine the
(C screen) and a suspended screen (V) impact on cassette optimum working parameters for this specific product. A summary of the performances of Pellicon® 3 cassette
resistance to fluid flow. The IgG solution was concentrated as much as possible according to the with D screen in comparison with C screen is presented in
following process strategy: Table 2.
Taking into account the typical maximum inlet pressure
for module, by simple interpolation, the maximum protein • Maintain the TMP set-point as long as possible by opening the back
concentration achievable for a process utilizing a V screen pressure valve until retentate pressure reached 2 – 3 psig;
device is expected to be much higher than C screen. When Product Attribute Performance Outcome
• Next, maintain the feed flow rate set-point as long as possible until a
dealing with materials at a high concentration/viscosity, the
max Pinlet = 60 psig is reached; Final viscosity > 40 cP with IgG
selection of the most appropriate screen for the UF cassette is
critical for process success. • Slowly ramp down the feed pump to maintain Pfeed at the max Pinlet
= 60 psig until the minimum achievable flow is reached (the pump Feed channel DP - proteins 50% of C screen at ≥ 25 cP
cannot be turned down any further) or the flux drops to zero;
140.0 Mass transfer k ≥ 75% of C screen
V Screen
120.0 C Screen Max protein concentration ≈ 1.25 X of C screen
100.0
Membrane area ≈ 1.25 X of C screen
Figure 5 shows the flux vs. concentration curves for the various
80.0
dP (psi)

screens. By the stagnant film model, the slopes represent the mass
60.0 transfer coefficient for each device. Table 2. Performances comparison D screen vs.
40.0 C screen
Flux vs. concentration at different feed flow Figure 5:
20.0 90
D screen 3 LMM Cassettes From internal and external experience, the D screen
80
0.0 D screen 5 LMM permeate is suitable for high concentration TFF ultrafiltration
0.0 50.0 100.0 150.0 200.0 250.0 70 D screen 7 LMM
applications and can process 2 to 3 higher viscosities than
Concentration of BgG (g/L)
C screen 3 LMM flux vs. IgG
60 C screen 5 LMM the C screen at typical process cross flow rates.
concentration at
Flux (LMH)

50 C screen 7 LMM

Figure 2: C and V screen modules resistance 3 different feed In addition, the D screen is consistently able to achieve
40 4-25% higher final product concentrations compared to
versus protein concentration flows
30 the C screen at the processing limits. It showed a 0-24%
20 reduction in mass transfer compared to the C screen. In
MilliporeSigma, Pellicon and Milli-Q are registered trademarks of addition, the D screen required 5-25% more membrane
Merck KGaA, Darmstadt, Germany. 10
Lit. No. PS6423EN00 Ver. 2.0 0
area (or process time) compared to the C screen for a
Copyright © 2017 EMD Millipore Corporation. All Rights Reserved. 10 100 1000 similar process (Significant improvement over V screen
Concentration (g/L) which requires 2 times more area). 
The life science business of Merck KGaA, Darmstadt, Germany
operates as MilliporeSigma in the U.S. and Canada.