Introduction to filter

sizing
Vmax and Pmax method
Presentation Overview

 How a Filter Works

 Key Membrane and Depth Filter Characteristics

 Filter Fouling Mechanisms: An introduction to Filter Sizing and

Scaling Models

 Gradual Pore Plugging Model and Vmax method

 Pmax/Tmax Method

 Safety Factor consideration

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How a Filter
Works
 Microporous Membranes

Virus Membranes

Ultrafiltration Membranes

Reverse Osmosis Membranes

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Retention Mechanisms

 Describe how filters capture (retain)

particles
 Mechanisms can be affected by:
• Fluid characteristics
• Operating conditions
• Particle type
• Filter type
 Retention mechanisms form the
foundation of filter fouling models

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Particle Retention Mechanisms

 Size Exclusion - Plugging

• Sieving (surface)
• Entrapment (depth)
• Size-dependent

 Adsorption
• Attraction forces between particles
and filter material
• Molecular and/or electrical
• Not size-dependent

 Depends on
• Particle type
• Solution properties
• Filter material and structure
Key Membrane and
Depth Filter
Characteristics
What do Membrane Filters look like?
 Mainly made by casting membrane
 Can be either hydrophilic or hydrophobic
 Rated on the size of the smallest particle it
retains
 Very thin (100 - 260 um)
 Adsorption depends on materials
• Not the primary retention mechanism
 Examples
• Cellulose ester
• Regenerated cellulose
• Nylon
• Polysulfones
• PVDF

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Key Membrane Filter Characteristics

 Strong, Rigid NOT brittle

 Tortuous pathway
 Very high internal area
 65-75% porosity
 Size exclusion - particle
retention does not change
with flow or pressure
 Sterilizing filters must have
> 99.99999% removal
and sterile filtrate
 Integrity testable (diffusion &/or bubble
point)

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What do Surface (Pre-) Filters look like?

 Fibers locked together by heat or

membrane coating
 Given a nominal rating or rated by the filter
it protects
 Thin (1 mm or less) & Slightly Adsorptive
 Give a percentage (90 - 99.9%) particle
reduction
 Examples
• Cellulose ester coated cellulose or
polyester web
• Heat-treated polyproplylene filters

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 What do Depth Filters look like?

 Fibrous (can shed fibers)

 Difficult to give an accurate pore size
rating
 Thick (3 - 30 mm) & often adsorptive
 Give a percentage (i.e. 30 - 70%)
particle reduction
 Have the greatest capacity
 Examples
• Microfiberglass
• String-wound filters
• Sheet / pad filters

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 Depth filter composition

 Cellulose fibers base matrix

• Highly refined
 DE (Diatomaceous Earth) Diatomaceous Earth

• Refined grade
• Large surface area
• Entrapment sites
 Resin binder
• Positive charge & hydrophobicity
for adsorption

Filter Matrix Cross-Section @ 3656 X

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Filter Fouling
Mechanism:

An introduction to filter
sizing
Types of Particles in Biological Fluids
 Non-deformable types
• Resin beads or fines
• Drug crystals
• Carbon fines
• Diatomaceous Earth (D.E.)
• Form porous permeable
cakes.

 Deformable types
 Proteins

 Lipids

 Sugar/protein complexes

 Can move through the filter,

break-up and compress into

impermeable cakes.
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Cake Formation

 Happens with hard particles

 Particles build up on the surface of the
filter
 If particles are rigid, resistance
increases linear with cake thickness

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Complete Pore Blocking

 Happens with deformable particles

 Pressure forces particles to completely
block the "pore"
 Common when there is poor or no
prefiltration OR when soft particles
slightly larger than the filter rated pore
size

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Gradual Pore Plugging

 Happens with hard or deformable

particles
 Particles build up on the "pore" throat
or opening
 Filter slowly blocks
 Most common with biological fluids

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Impact on Filter Behavior
 Gradual blockage most common

 "Everything was going alright,

then all of a sudden the filter
plugged"

Constant flow
filtration –
∆p increases as
filter fouls
Gradual and complete blocking do
not have a linear relationship
between ΔP and capacity
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Filter Performance Characterization

Filter performance is defined by two key attributes

Capacity
• Volume that can be process per filter area (L/m2)
• How much?
 Flowrate
• Volume processed per time per area (L/m2/hr = LMH)
• How fast?
Performance depends on:
• Filter selection- the correct filter for the application
• Process parameters
• Optimizing pressure, flowrate, time, area

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Small-Scale Test Methodologies
Constant Pressure (Vmax)
 Fluid is held at constant
pressure and forced through
filter media
 Filter plugging is observed by
the decrease in flow rate over
time
 Classically, based on
gradual pore plugging
model
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Constant Flow Rate
(Pmax/Tmax)
 Fluid is pumped at a constant
flow rate through the filter
media
 Filter plugging is observed by
an increase in differential
pressure over time
OR
 Filter plugging is observed by
an increase in filtrate turbidity
over time
 Based on a small-scale
process simulation with
empirical data fitting
Choosing a Filter Sizing Technique:
Fouling Mechanism Basis

Constant
Sizing Method Name:
Mode of Testing

Pressure Vmax
Volume Endpoint Vmax

Constant Tmax Pmax Pressure Endpoint Pmax

Flow
∆P does ∆P increases during Turbidity Endpoint Tmax
not change filtration

Size Exclusion Mechanisms

Choosing a Filter Sizing Technique:
Filter Type Basis

Tmax

Pmax Pmax (Applicable,

(Application Focus) but less common)

Vmax

Depth Membrane and Non-woven Sterilizing-grade

Filters Prefilters Membrane Filters

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Gradual Pore
Plugging
and Vmax
Method
Available Test Methodologies for Sizing Filters
Constant Pressure (Vmax)
 Measures decrease in flow as a
function of throughput
 Endpoint is determined by flow rate or
volume
flow rate
throughput
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Constant Flow Rate (Pmax)
 Measures increase in pressure and
decrease in filtrate quality as a
function of throughput
 Endpoint is determined by pressure
limit or desired filtrate quality
pressure
turbidity
throughput
Filter Plugging Models

Mechanism of filter plugging:

• d2t/dV2 = k(dt/dV)n
where:
t = filtration time
V= cumulative volume at time t
k = constant whose dimensions are dependent on n
n = 1.5 for gradual plugging

• H.P. Grace, "Structure and Performance of Filter Media,"

AICHE Journal 2(3), 307-336 (1956)
In practical terms:

• t/v = t/Vmax + 1/Qi

where:
Vmax = maximum volume that can be filtered at time infinity
Qi = instantaneous initial flow

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The Vmax (Constant Pressure) Test
• Accelerated screening technique to estimate scaled-up filter size requirements
• Helps to optimize filtration train rapidly

• Estimates the maximum fluid volume filterable

through a filter
– Predicts Capacity, Vmax [=] L/m2 (@ t=∞)
– Predicts Flux Decay Profile , Q [=] L/min
• Vmax Characteristics
– Based on the gradual pore plugging model
• Vmax Implementation
– Plot t/V versus t at constant ∆P
– Vmax = 1/Slope, Qi = 1/Intercept

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Vmax: Result Analysis

 Typical Curve
• Highly linear region r2 > 0.99
− r2 > 0.99 1/Qi
t/V 1/Vmax
 What happens when r2 < 0.99?
• Prediction of Capacity (Vmax) based on
10 min test becomes less reliable
• Remove earlier points to see if fit is
t
improved,
− Need at least 6 points in the
straight line for reliable correlation r2 < 0.99

 Run test to 80% plugging (flow t/V

decay) 1/Vmax

t
Vmax: Approaches to Filter Sizing

Three process scenarios or cases are usually relevant:

• Case 1: Batch Volume of fluid to be filtered is given
• Case 2: Batch Volume of fluid to be filtered at a maximum allowable
process time is given
• Case 3: Batch Volume of fluid to be filtered with a specified minimum
allowable flow rate is given

Largest surface area that fulfills all process requirements is selected, Amin

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Vmax: Sizing Equations

Case 1. Only VB (Batch Volume is given; No batch time, minimum flow)

VB
Amin =
Vmax
• Eq. gives the minimum area required (no safety factor is included)
• Ensure that Amin leads to respectable batch times
Case 2. VB (Batch Volume) and tB (Batch time) are given
VB VB
Amin = +
Vmax Qi × t B
Case 3. VB (Batch Volume), tB (Batch time) & Qmin (minimum flow rate) are given

Qmin VB
1− =
Qi × Amin Vmax × Amin
• Using an Iterative Method, Solve Eq. For ‘Amin’
•‘Case 2’ can give a reasonable initial guess
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Vmax: Advantages/Benefits

Simple, rapid & easy to use

Experimental basis for filter train selection
• Establishes optimized filtration train and preliminary information for
scale-up, confirmatory pilot scale trials
Results have simple interpretations:
• Vmax - maximum ‘filterable’ fluid volume before plugging, L/m2
• Qi - initial filtrate flowrate; L/min/m2
Requires only limited fluid volume to perform the test.
• < 1 liter

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Vmax: Limitations

It does not tell you which filter to test

• Experience & historical records are useful
Does not tell you anything about filtrate quality
• Indirectly Vmax, with a tighter filter, on the filtrate is a measure of filtrate
quality
Does not simulate the entire process
• Need for intermediate pilot trials
Only applies to gradual pore plugging model

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Pmax/Tmax
method
Large scale  Small scale

Golden rule:

“ During PD, in small scale experiments,

mimic large scale operation
as close as possible”

 Process parameters
 Fluid characteristics

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Challenges to Clarification
Process Optimization

Three process challenges when developing a clarification scheme:

1. Understanding fluid complexity and characteristics
2. Understanding how to select optimal separation technology
3. Integrating clarification technologies to achieve the optimal clarification
scheme

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First Process Challenge: Understanding Fluid Complexity

Biological Fluid Constituents

– Solids (> 1 µm)
– Soluble entities
• Easily removed by depth entrapment,
• Protein
cake build-up or centrifugation
– Cell culture fluids
– Cells and organisms (0.2 - 50 µm)
– Lysates
• Mammalian cells
– Plasma fractions
– Delicate, can lyse under stress
and release plugging materials • Salts

• Others – Buffers

– Bacteria, yeast, insect, plant • Preservatives

– Colloids (0.01 - 1.0 µm) – Ophthalmics

• Deformable and “sticky” • Anti-foams

• Usually negatively charged – Cell culture fluid

• Plug downstream steps • Plant hydrolysate& serum

– Cell culture media

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Second Process Challenge: Optimal Technology Selection

What Technologies Are Available?

Separation Application
Benefits Limitations
Technology Scope

Simple scale up and

No filter re-use, High
Clarification, implementation, Multiple media
NFF expendables, Limited by solid
Prefiltration options (Surface, depth,
load
charged)
Clarification, Efficient and robust process,
High capital cost, Difficult to
TFF Concentration, Can handle high solid loads,
operate/validate
Cell recovery Re-usable filters
Cheap operation, No
Mechanical Clarification High Capital cost, Source of
expendables, Robust, Re-
Separators Only yield loss, Difficult scaleablility
usable, Easy to clean

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 Third Process Challenge: Optimizing Technology Integration

Cell Density
TFF or centrifuge
High TFF
+ 1-stage charged
TFF or centrifuge depth filter
+ 2-stage charged
depth filter
Med Depth filter
1 or 2-stage
charged
Low TFF or centrifuge depth filter
+ 1-stage charged Surface filter
depth filter
Cell Viability

Low Med High

Integrating Multiple Technologies Will Achieve

The Optimal Clarification Scheme
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Filter Selection Rules of Thumb

Fluid Fluid Components/ Recommended Filters

Turbidity Characteristics

< 20 NTU Colloids, small Protected sterile filter (0.45/0.22 µm

particulates Durapore, Express SHC)

20-100 NTU Colloids, small Membrane, cartridge-style pre-filter

particulates (Milligard, PolySep II)

100-300 NTU Colloids, cell debris, Smaller pore depth filter

particulates (X0HC, A1HC, B1HC)
(2nd clarification)
> 300 NTU Whole cell, hard Larger pore depth filters
particles, cell debris (D0HC, C0HC, DE50)
(1st clarification)

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Capacity

Goal:
• Maximize the amount of fluid that can be processed through a filter.

The question is:

"At what point does the filter resistance (pressure) or filtrate quality
negatively impact the process?"
• excessive pressure
• excessive turbidity breakthrough

Capacity
The volume filtered up to the point when the maximum pressure or
turbidity is achieved
• defined in volume/area (L/m2)

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The Pmax/Tmax (Constant Flow) Test

Using a peristaltic (or other positive displacement) pump, a constant flowrate is

maintained

Process simulation technique (scale-down)

Estimates the maximum fluid volume filterable through a filter

− Pressure Limited Capacity (Pmax)
− Filtrate Quality Capacity (Tmax)

Calculate resistance as a function of volumetric throughput

Sizing is based on an empirical model

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Methodology: Pmax
constant flow, P limitation

Constant flow filtration – pressure increases as pores are blocked

Flow rate through membrane will depend on installed area

Maximal pressure

Pressure
(psi)

Time(h) ~Volume (L)

Area of filter 1 < Area of filter 2

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Methodology : Pmax

 Maximum pressure P and

process time & volume
defined as end point.
Pressure
(psi)
 Define resistance
(psi/LMH) as pressure per
flow rate (J) per area Time(h) ~Volume (L)

 Define throughput as
volume per area (L/m2)

Resistance Pmax (L/m2)

(psi/(L/m2/h))

Throughput (L/m2)

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 Analyzing Test Results

Fit the data to a 2nd or 3rd order

polynomial
− Choosing the “best fit” comes with
experience—it is a judgment call
− In general:
− Polynomial must have the correct
concavity when projected forward
− Polynomial must accurately model
the “pressure rise” region

Use the Pmax sizing algorithm for a

known batch volume (VB), batch time (tB)

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 Applying the Pmax Sizing Algorithm

Pmax Sizing Algorithm: • Example Sizing (VB=1000L tB = 4 hr):

1. Guess an area & select ∆Plimit Fig. 1 - Resistance vs. Normalized Volume

for process endpoint Resistance

Permeate NTU
0.70 1.2

2. Calculate Flux, Javg = VB/AtB 0.60

1
y = 2.20E-08x3 - 6.13E-06x2 + 7.44E-04x - 2.53E-02

Resistance (psi/LMH)
and Capacity = VB/A 0.50

Permeate NTU
0.8
0.40

0.30 0.6
3. Calculate Resistance, Rcalc. = 0.20

∆P/Javg.
0.4
0.10
0.2
0.00
0 50 100 150 200 250 300 350 400 450
4. Determine Actual Resistance -0.10
Capacity (L/m²)
0

from the graph or Polynomial 1. Guess A=3.0 m2, Plimit = 15 psid

fit, Ractual
2. Javg = 83 LMH, V/A = 333 L/m2
5. Check to see if Rcalc = Ractual 3. Rcalc = 15/83 = 0.18 psid/LMH
• If yes, selected A is correct
4. Ractual = 0.36 psid/LMH
• If Rcalc < Ractual, increase
5. Rcalc < Ractual ∴ increase A & reiterate
Area (A) & iterate until Rcalc
= Ractual
• Amin = 3.5 m2
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Pmax/Tmax: Advantages/Benefits

Applicability is independent of the plugging model

− Uses an empirical data analysis method to analyze constant flow data
− not based on any mathematical model

Results have simple interpretations and provide basis for implementation

(process simulation):
− Pmax
− maximum ‘filterable’ fluid volume before pressure limit, L/m2
− Tmax
− maximum ‘filterable’ fluid before filtrate quality limit, L/m2

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How Accurate is the Pmax
Sizing Method?

 Pmax can be very

accurate when
process parameters

resistance (LMH/psi)
between large and
small-scale are
carefully controlled
 Example at right
shows a 130X scale-
up/scale-down
filter loading (l/m2)

Millistak Mini (2X flux) Millistak Mini (1X flux)

Millistak Pod (1X flux) sizer prediction
Pmax/Tmax: Limitations

• Uses an empirical data analysis method to analyze constant flow

data—not based on any physical model

• Requires longer test times

– Close to process times
– No extrapolations to higher loadings

• Checks need to be put in place to guarantee adequate fit of

resistance curve

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Safety Factors
Considerations
Safety Factors:
Introduction

General rule is 1.5X but…

It depends!
 Amount and quality of data
 Expected variability of feed
 Expected variability of filter
 Existing equipment
 Scaling factor

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 Safety factors

“Variability is typically normal”

To allow for variability in your process, a safety factor is applied.

Depending on your unit operation and information on typical variability, this safety factor should be
smaller or larger.
- i.e. control over cell culture consistency, centrifuge operation can reduce required safety factor
and make secondary clarification very economical, lack of control might lead to large installations.

Risk/cost based approach

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 Safety Factors:
Using a cost benefit analysis

 Analysis based on a sample

of sterile filters $10,000
$9,000
 Lower area yields lower $8,000 fail batch
overall cost but increases $7,000 yield loss
risk of a failed batch $6,000

$/batch
labor
$5,000
total buffer
 Higher membrane area $4,000
capital
$3,000
increases overall cost but $2,000
consumable

risk of lost batch is $1,000

decreased $-

4" Opti
MPAK20
MPAK40

MPAK60
MPAK100

MPAK200

1
2
3
4
5
6
7
8
9
12

15
18

21
38
# 10" elements

• Existence of a minimum value for a failed batch vs. overall cost is the base case
for determining a safety factor
Safety factor:
Choosing a robust safety factor
1000.00

Each unit operation has its

Safety 1.1
Factor
1.2
own range of 1.4
100.00
• Cost ratios Sterile Bulk Clarification 1.6
1.8
or media or Virus
• Variability

Cost Ratio
2
10.00 2.2
Similarly, each unit 2.4

operation has its own range Sterile 2.6

Buffer
of safety factors
2.8
1.00
3

“Standard” safety factors 3.2

3.4
may be insufficient 0.10
0% 10% 20% 30% 40% 50% 60%
Coefficient of Variation of Area

Cost of batch Safety

Application failure Area Variable cost Cost ratio COV Factor
Final Bulk sterilization $500-1500K 1-7 sqm $0.4-0.6 K/sqm 400-800 10-30% 1.4-2.0
Buffer sterilization $5-50K 1-7 sqm $0.4-0.6 K/sqm 5-25 5-10% 1.1-1.3
Growth media
sterilization $50-500K 1-7 sqm $0.4-0.6 K/sqm 50-250 10-30% 1.3-2.0
Clarification $100-1000K 1-30 sqm $0.3-0.5 K/sqm 50-250 30-60% 1.8-2.8
Virus filtration $500-1500K 1-7 sqm $2.8-7.5 K/sqm 30-250 40-60% 2.0-3.0

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Safety Factors for Depth Filtration

Variable Failure Safety

Application $/g $/g CR COV Factor
Sterile Bulk 0.03-0.10 100-300 1000- 15-20% 1.6-1.9
Buffer 0.2-0.4 1-2 2-10 15-20% 1.3-1.5
Media 0.6-2.1 10-100 15-50 10-30% 1.3-1.9
Series 0.1-0.3 100-300 3000-7500 15-20% 1.6-1.9
Clarification 1.4-5.3 20-200 5-150 25-40% 1.5-2.3
Retrovirus 0.4-0.7 100-300 150-750 15-25% 1.5-1.9
Parvovirus 3.2-32.0 100-300 3-90 25-35% 1.4-2.1
UF 0.1-0.3 100-300 300-3000 5-25% 1.2-2.1

• Depth filtration has a relatively high cost ratio due to a relatively high variable
cost ($/g)
• The cost-benefit analysis suggests that safety factors of 1.5-2.3 are justified
• Historically, safety factors as high as 3.0 have been employed in depth filtration
operations (depends on control over feed variability)
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 Wrap-Up

Good filter sizing starts with well-designed small-scale trials

− Multiple experiments
− The right test methodology
− Proper understanding of sizing techniques
Intermediate (pilot-scale) trials will ensure smooth scale-up
Implementing the “right” filter involves more than capacity; other
considerations include:
− The right safety factor
− Process limitations
− Hardware constraints
− SIP requirements
− Robust IT operations

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