validation guide · results of extractables level vs. volumetric throughput of purified water...

Doc. No. VGWA2012REV1.0

VALIDATION GUIDE

for WaterSep Hollow Fiber Cartridges

2 WaterSep Validation Guide

Terms of Sale

WaterSep sells all goods and services per the terms and conditions of sale as specified by the WaterSep sales agreement.

To receive a copy of these terms and conditions, or to comment on our products, contact us at:

WaterSep Technology Corporation

420 Maple Street, Suite 1

Marlborough, MA 01752

USA

Telephone: +1–508–970–0089, extension 204

Fax: 508-970-0146

Email: [email protected]

All third party trademarks are the property of their respective owners.

© 2013 by WaterSep Bioseparations. All rights reserved.

WaterSep Validation Guide 3

CONTENTS

List of Tables 5

List of Figures 5

Chapter 1—Overview

How this Validation Guide Can Help You 6

Intended Audience 6

Getting Help 7

What is Process Validation? 7

Chapter 2—Product Information

Lab-, Pilot- and Production-Scale Hollow Fiber Cartridges 9

Intended Applications 9

Cartridge Design 10

Cartridge Labeling and Catalog Numbers 11

Materials of Construction 14

Packaging 14

Product Specifications 15

Void Volume of Hollow Fiber Cartridges 15

Membrane Performance 20

Cartridge Performance 20

Storage of New and Used Cartridges 22

Preparing Your Cartridge for Use 22

Chapter 3—Validation Information

Membrane Water Flux 23

Membrane Solute Rejection 24

Cartridge Integrity Test 25

Non-Destructive Testing 25

Integrity test - Pressure Hold Test 25

Reference Values for WaterSep Factory Membrane diffusion test 27

Cartridge Water Flux 27

Cartridge Crossflow 30

Cartridge Chemical Compatibility Study 33

Cartridge Rinsing and Buffer Equlibration Study 35

Hollow Fiber Cartridge Scalability Studies 36


Chapter 4—Product Safety

Biocompatibility Studies 39

Chapter 5—Quality Assurance Information

Certificate of Compliance 40

Appendices

Appendix I—Additional Documentation 41

Appendix II—Test Procedures and Reports 42

Index

Index 43


LIST OF TABLES

Table 1. Overview of the hollow fiber cartridges described in this validation guide 9

Table 2. Physical characteristics of MiniDiscover Green, Discovery Green, and Explorer

Green HF Cartridges 16

Table 3. Physical characteristics of Investigator and BioProducer cartridges 17

Table 4. Void volumes of WaterSep Green hollow fiber cartridges 18

Table 5. Air flow specifications for WaterSep Green single use hollow fiber cartridges 21

Table 6. Water permeability of WaterSep membranes 23

Table 7. Results of solute passage through WaterSep membrane (% P – percent passage)

24

Table 8. Results of membrane integrity tests for Investigator12 Cartridges 27

Table 9. Water flux results for an Investigator12 hollow fiber cartridges 28

Table 10. Chemical compatibility list for WaterSep cartridges (R = recommended, L =

limited exposure, NR = not recommended, U = unknown) 33

Table 11. Results from the E. Coli lysate clarification scalability study 37

Table 12. Helpful information available at www.WaterSep.net 41

Table 13. Test reports available from www.WaterSep.net 42

LIST OF FIGURES

Figure 1. Main parts of a WaterSep hollow fiber cartridge 10

Figure 2. Cartridge showing hollow fibers encapsulated in the hollow fiber housing 11

Figure 3. Typical hollow fiber cartridge label show information helpful to users 12

Figure 4. Identifying cartridge properties by catalog number 13

Figure 5. Clean water flux for an Investigator12 Green, 30K MWCO, 1.0 mm ID 28

Figure 6. Clean Water flux for an Investigator12 Green, 300K MWCO, 1.0 mm ID 29

Figure 7. Clean Water flux for an Investigator12 Green, 0.2 um, 1.0 mm ID 29

Figure 8. Delta P versus crossflow with water for Producer12/24/41 Green cartridges 31

Figure 9. Delta P versus crossflow with water for Investigator12/24/41 Green cartridges 32

Figure 100. Results of Extractables Level vs. Volumetric Throughput of Purified Water

(Liters per m2) 35

Figure 111. Pressure and permeate flux profiles for the Explorer12 cartridge 36

Figure 122. Pressure and permeate flux profiles for the Explorer24 cartridge 37

Figure 133. Optimization results of scalability study using Explorer12 and Investigator12

cartridges 38

Figure 144. Example of a cartridge certificate of compliance 40

Overview


CHAPTER 1—OVERVIEW

HOW THIS VALIDATION GUIDE CAN HELP YOU

WaterSep created this validation guide to help scientists and engineers use our hollow

fiber cartridges properly and efficiently. You can benefit many ways from reading

and understanding the information in this validation guide:

You can save considerable time when setting up and using your hollow fiber cartridge.

You can obtain consistent results and extend the service life of your cartridge.

You can find the information you need to help validate your hollow fiber cartridge

system to meet FDA regulations.

WHAT YOU WILL LEARN

Designing and validating a hollow fiber cartridge system to meet FDA requirements

involves applying technical knowledge in an organized fashion. While the path to system

design and validation can take many directions, the performance and specifications of the

hollow fiber cartridge remain constant. You will need the following information to

validate your WaterSep cartridges within your application:

Product labeling, materials of construction, cartridge physical characteristics, and

product performance specifications

Results of our cartridge integrity, performance, compatibility, and scalability studies

Results of extractable, USP, hemolysis, and cytotoxicity tests performed by

independent testing laboratories

Operational, quality control, and regulatory support documents

INTENDED AUDIENCE

This validation guide was written for scientists and engineers who have laboratory

operation and process engineering skills. If you need assistance, or do not fully

understand the information in this guide, contact WaterSep Bioseparations for support.

Overview


GETTING HELP

WaterSep engineers are membrane filtration experts. If you have questions or need

specific product or application information, please contact our technical support team.




USA

Telephone: +1–508–970–0089 x204

Fax: 508–970–0146

Email: [email protected]

WHAT IS PROCESS VALIDATION?

If you are new to process validation, the FDA offers the following introduction as quoted

from CPG Sec. 490.100 Process Validation Requirements for Drug Products and Active

Pharmaceutical Ingredients Subject to Pre-Market Approval.

"Validation of manufacturing processes is a requirement of the Current Good Manufacturing Practice (CGMP) regulations for finished pharmaceuticals (21 CFR 211.100 and 211.110), and is considered an enforceable element of current good manufacturing practice for active pharmaceutical ingredients (APIs) under the broader statutory CGMP provisions of section 501(a)(2)(B) of the Federal Food, Drug, and Cosmetic Act. A validated manufacturing process has a high level of scientific assurance that it will reliably produce acceptable product. The proof of validation is obtained through rational experimental design and the evaluation of data, preferably beginning from the process development phase and continuing through the commercial production phase. Refer also to the Guideline of General Principles of Process Validation (May 1987, originally published by CDER, CBER, and CDRH and presently recognized by CDER, CBER, and CVM)1. (Note: The guideline is under revision as of the date of this CPG.) Before commercial distribution begins, a manufacturer is expected to have accumulated enough data and knowledge about the commercial production process to support post-approval product distribution. Normally, this is achieved after satisfactory product and process development, scale-up studies, equipment and system qualification, and the successful completion of the initial conformance batches. Conformance batches (sometimes referred to as "validation" batches and demonstration batches) are prepared to demonstrate that, under

Overview


normal conditions and defined ranges of operating parameters, the commercial scale process appears to make acceptable product. Prior to the manufacture of the conformance batches the manufacturer should have identified and controlled all critical sources of variability."

Guidelines for validation of biological systems and processes can be found in publications

from the FDA.

This validation guide provides you with information that can facilitate validation of your

WaterSep products and systems. In addition, WaterSep can provide you with additional

membrane and product information specific to your particular needs. For assistance,

contact WaterSep at +1–508–970–0089, extension 204.

As you begin to validate your system, it is helpful to know that you must accomplish

three qualifications: 1—An installation qualification (IQ) that verifies that the hollow fiber

membrane process and supporting equipment can consistently operate within established

limits and specifications. 2—An operating qualification (OQ) that verifies that the process

can consistently reproduce operating results and that the process is effective.

3—A performance qualification (PQ) that verifies, through operational testing and data,

that the finished product produced by the specific process meets and conforms

to the product release specifications for functionality and safety.

Product Information


CHAPTER 2—PRODUCT

INFORMATION

LAB-, PILOT- AND PRODUCTION-SCALE HOLLOW FIBER

CARTRIDGES

WaterSep manufactures hollow fiber cartridges for lab-, pilot- and production-scale

separation operations (Table 1). The cartridges are designed for linear scale-up, offering

consistency in design elements such as materials of construction, fiber configuration, and

fiber length. Other product benefits include low fouling performance and easy cleaning.

This Validation Guide covers WaterSep’s Green Line, the single use glycerin free hollow

fiber product line.

Table 1. Overview of the hollow fiber cartridges described in this validation guide

Cartridge Type Applications Range of

Surface Areas

Range of Sample Volumes/Cartrid

ge Lengths (Inches)

MiniDiscovery Green Lab 0.019–0.038 ft2 (17.3–35.6 cm2)

10–125 ml 12 and 24

Discover Green Lab 0.056–0.115 ft2

(51.8–106.9 cm2) 10–400 ml 12 and 24

Explorer Green Lab 0.16–0.62 ft2

(155–580 cm2) 150–3000 ml 12, 24, and 41

Investigator Green Pilot 1.4–5.5 ft2

(0.13–0.50 m2) 1–25 L 12, 24, and 41

BioProducer Green Production 13.5–54 ft2

(1.25–5.0 m2) 10–300 L 12, 24, and 41

Maximizer Green Production 54–110 ft2

(5.0–10.0 m2) > 300 L 24 and 41

INTENDED APPLICATIONS

The intended applications in the biopharmaceutical/bioindustrial sectors include

crossflow microfiltration and ultrafiltration for:

Primary recovery/clarification

Vaccine purification and

concentration

Cell washing

Cell harvesting

Bacteria separation

in fermentation broths

Debris removal post-centrifugation

Primary recovery of recombinant

biopharmaceuticals

Macromolecule concentration

and diafiltration (enzymes, antibodies,

proteins, and viruses)

Product Information


CARTRIDGE DESIGN

Hollow fiber cartridges consist of a housing with an inlet port for feed, an outlet port

for retentate, and two outlet ports for the permeate (Figure 1).

The housings contains hollow fiber membrane made from an antifouling modified

polyethersulfone (PES) composition (Figure 2).

Fluid flows into the feed port, through the lumens of the hollow fibers and out of the

retentate port, returning to the feed tank. Fluid and solutes that pass through the walls

of the hollow fiber membrane are flow out the two permeate ports.

Cartridge size, port size, port type, and the number and size of hollow fibers vary

according to the cartridge type.

Figure 1. Main parts of a WaterSep hollow fiber cartridge

Outlet (retentate)

Inlet (Feed)

Outlets (permeate)

Housing

Product Information


Figure 2. Cartridge showing hollow fibers encapsulated in the hollow fiber housing

CARTRIDGE LABELING AND CATALOG NUMBERS

WaterSep hollow fiber cartridges include labels that provide useful information (Figure 3).

Cartridge labels include the following information:

Company name

Cartridge name

Membrane material and pore size

Inside diameter of the fibers

Membrane surface area

Cartridge nominal dimensions

Bar code

Maximum operating temperature

Maximum operating pressure

Catalog (part) number

Lot number

Encapsulated fibers Housing

Product Information


When ordering cartridges, the catalog number enables you to identify the cartridge

properties (Figure 4). The bar code enables you to identify each cartridge using a universal

bar code scanner. The lot number is also unique to each cartridge to ensure traceability.

Figure 3. Typical hollow fiber cartridge label show information helpful to users

Product Information


Figure 4. Identifying cartridge properties by catalog number

SU 910 05 DIS 12 LL

Product Code Designator

SU = Modified PES Membrane Glycerin Free

Fiber Lumen Diameter (mm)

20 = 2 mm (special production)

10 = 1 mm

05 = 0.5 mm

NMWC

001 = 1 K

003 = 3 K

005 = 5 K

010 = 10 K

030 = 30 K

050 = 50 K

100 = 100 K

300 = 300 K

500 = 500 K

750 = 750 K

910 = 0.1 µm

920 = 0.2 µm

945 = 0.45 µm

965 = 0.65 um

Cartridge Type

DIS = Discover Green

EXP = Explorer Green

INV = Investigator Green

PRO = BioProducer Green

MAX = Maximizer Green

Fiber Length

12 = 12 inches

24 = 24 inches

41 = 41 inches

Type of Connector

ML = Luer Lok (Mini Discover)

LL = Luer Lok (Discovery)

SO = 3/4-inch TC (Explorer)

SO = 1-inch TC (Investigator)

SG = 1.5-inch TC (BioProducer)

SH= 2-inch TC (Maximizer

SK = 3-inch TC (BioProducer)

Catalog number

Product Information


MATERIALS OF CONSTRUCTION

The materials of construction (and wetted components) of WaterSep hollow fiber

cartridges include these:

Membrane—modified glycerin free polyethersulfone (PES) composition

Encapsulant—FDA-compliant urethane

Housing—white polysulfone

Polypropylene Screen—made from 100% virgin polypropylene resin.

PACKAGING

New hollow fiber cartridges are packaged in a sealed polyethylene bag. The feed,

retentate, and permeate ports are double capped to retain moisture and prevent

contamination from external sources.

Depending on the cartridge size, packaging consists of different types of foam and

cardboard boxes. A label—similar to the label affixed to the cartridge—is affixed to the

upper-left side of each box.

The finished goods package also includes a certificate of analysis, operating instructions,

and an application guide.

Product Information


PRODUCT SPECIFICATIONS

You can purchase WaterSep Green Single Use hollow fiber cartridges in a variety of sizes

from lab scale to production scale (Table 2 and Table 3). By design, WaterSep Single Use

hollow fiber cartridges provide a linear and predictive scale-up process—from laboratory

to pilot-scale to manufacturing scale—by using matching materials, fluid-path length, and

performance characteristics. A typical scale-up path includes these products and volumes:

PRODUCT VOLUME

MiniDiscover/Discover Green 10–400 ml

Explorer Green 150–3000 ml

Investigator Green 1–25 L

BioProducer Green > 10 L

Maximizer Green > 300 L

For additional scale-up information, contact WaterSep at +1–508–970–0089,

extension 204, or email WaterSep at [email protected]

VOID VOLUME OF HOLLOW FIBER CARTRIDGES

The design of WaterSep hollow fiber cartridges maximizes surface area and minimizes

hold-up volume and system working volume. The design elements increase efficiency and

enable the highest possible concentration factors.

Void volumes and other product characteristics for MiniDiscover/Discover, Explorer,

Investigator, BioProducer Green and Maximizer Green hollow fiber cartridges are

presented in Table 2, Table 3, and Table 4.

Product Information


Table 2. Physical characteristics of MiniDiscover Green, Discovery Green, and Explorer

Green HF Cartridges

MiniDiscover Green HF Cartridges

Characteristic 12 Inch 24 Inch —

Dimensions in inches (mm) 3/8 x 12 (9.4 x 30) 3/8 x 24 (9.4 x 60) —

Membrane surface area in ft2 (cm2) 0.019 (17.3) 0.038 (35.6) —

Recommended batch volume per cartridge (mL) 10–80 10–125 —

Recommended permeate flow rate (mL/hr) Up to 70 Up to140 —

Membrane cut-off (k) NMWC 1, 3, 5, 10, 30, 50, 100, 300, 500, 750

Membrane pore size (µm) 0.1, 0.2, 0.45, 0.65

Feed/retentate connectors Luer Lok

Permeate connector Luer Lok

Discover Green HF Cartridges

Characteristic 12 Inch 24 Inch —

Dimensions in inches (mm) 3/8 x 12 (9.4 x 30) 3/8 x 24 (9.4 x 60) —

Membrane surface area in ft2 (cm2) 0.056 (51.8) 0.115 (106.9) —

Recommended batch volume per cartridge (mL) 10–250 50–400 —

Recommended permeate flow rate (mL/hr) Up to 200 Up to 400 —



Feed/retentate connectors Luer Lok

Permeate connector Luer Lok

Explorer Green HF Cartridges

Characteristic 12 Inch 24 Inch 41 Inch

Dimensions in inches (mm) 0.5 x 12.3 (13 x 312) 0.5 x 23.8 (13 x 605) 0.5 x 41.8 (13 x 1062)

Membrane surface area in ft2 (cm2) 0.16 (155) 0.34 (320) 0.62 (580)

Recommended batch volume per cartridge (mL) 150–175 250–1,500 300–3,000

Recommended permeate flow rate (mL/hr) Up to 600 Up to 1,280 Up to 2,300



Feed/retentate connectors ½-inch TC

Permeate connector Barbed hose

Product Information


Table 3. Physical characteristics of Investigator and BioProducer cartridges

Investigator Green HF Cartridges


Dimensions in inches (mm) 1.3 x 12 (33.4 x 305) 1.3 x 23.5 (33.4 x 597) 1.3 x 41 (33.4 x 1054)

Membrane surface area in ft2 (m2) 1.4 (0.13) 2.3 (0.27) 5.5 (0.50)

Recommended batch volume per cartridge (L) 1–6 2–12 3–25

Recommended permeate flow rate (L/hr) Up to 5.2 Up to 11 Up to 20



Feed/retentate connectors 1-inch TC

Permeate connector ½-inch TC

BioProducer Green HF Cartridges


Dimensions in inches (mm) 3.5 x 15.0 (89 x 381) 3.5 x 13.5 (89 x 344)

3.5 x 26.5 (89 x 673) 3.5 x 25.0 (89 x 636)

3.5 x 44.5 (89 x 1130) 3.5 x 43.0 (89 x 1093)

Membrane surface area in ft2 (m2) 13.5 (1.25) 29.3 (2.72) 54.0 (5.0)

Recommended batch volume per cartridge (L) 10–60 20–140 50–300

Recommended permeate flow rate (L/hr) Up to 50 Up to 110 Up to 200



Feed/retentate connectors 1.5-inch TC or 3-inch TC

Permeate connector 1-inch TC

Maximizer Green HF Cartridges

Characteristic 24 Inch 41 Inch

Dimensions in inches (mm) 4.62 x 28.5

(117.3 x723.9) 4.62 x 46.5

(117.3 x 1181.1)

Membrane surface area in ft2 (m2) 54.0 (5.0) 110 (10)

Recommended batch volume per cartridge (L) > 300 > 300

Recommended permeate flow rate (L/hr) Up–200 Up–400



Feed/retentate connectors 2-inch TC

Permeate connector 1-inch TC

Product Information


Table 4. Void volumes of WaterSep Green hollow fiber cartridges

Product

Performance Properties Hold Up Volume

Fiber ID

(mm) No. of Fibers

Surface Area (cm2)

Feed Side Holdup Volume

(ml)

Permeate Side Holdup Volume

(ml)

MiniDiscover12 Green

0.5 4

17

0.34 2.42

1.0 2 0.57 2.13

2.0 1 1.04 1.69


0.5 4

36

0.56 4.93

1.0 2 1.03 4.35

2.0 1 1.96 3.35


0.5 4

64

0.92 8.87

1.0 2 1.75 7.82

2.0 1 3.39 6.01

Discover12 Green

0.5 12

52

0.81 6.19

1.0 6 1.51 5.33

2.0 3 2.93 3.87

Discover24 Green

0.5 12

107

1.49 12.79

1.0 6 2.89 11.04

2.0 3 5.68 8.04

Discover41 Green

0.5 12

193

2.57 23.12

1.0 6 5.04 19.96

2.0 3 9.98 14.56

Explorer12 Green

0.5 36

155

2.7 14.4

1.0 18 4.8 11.8

2.0 9 9.1 7.4

Explorer24 Green

0.5 36

321

4.8 28.0

1.0 18 9.0 24.2

2.0 9 17.3 15.2

Explorer41 Green

0.5 36

579

8.0 50.5

1.0 18 15.4 43.5

2.0 9 30.3 27.3

Investigator12 Green

0.5 320

1,316

20 94

1.0 160 39 79

2.0 80 76 41


0.5 320

2,785

38 204

1.0 160 75 171

2.0 80 150 93


0.5 320

5,083

67 376

1.0 160 133 315

2.0 80 265 173

Product Information


Product

Performance Properties Hold Up Volume

Fiber ID

(mm) No. of Fibers

Surface Area (cm2)

Feed Side Holdup Volume

(ml)

Permeate Side Holdup Volume

(ml)

BioProducer12 Green

0.5 3200

12,514

348 94

1.0 1600 537 79

2.0 800 914 41

BioProducer24 Green

0.5 3200

27,198

532 204

1.0 1600 904 171

2.0 800 1,647 93

BioProducer41 Green

0.5 3200

50,183

819 376

1.0 1600 1,478 315

2.0 800 2,796 173

Maximizer24 Green

0.5 6400 51,842

1,194 3,126

1.0 3200 1,937 2,502

2.0 1600 51,868 3,425 1,032

Maximizer41 Green

0.5 6400 97,811

1,768 5,701

1.0 3200 3,086 4,522

2.0 1600 97,837 5,722 1,769

Product Information


MEMBRANE PERFORMANCE

The WaterSep Green single use hollow fiber membranes are manufactured using a

unique combination of modified polyethersulfone (PES) and a proprietary spinning

process that results in an asymmetric, void free low binding hollow fiber membrane with

narrow pore size distribution and excellent antifouling and process flow properties.

The manufacturing process for the Green single use hollow fiber membrane is free of

glycerin or any other humectant, which guarantees a very low level of extractables.

CARTRIDGE PERFORMANCE

The exceptional uniformity of WaterSep hollow fiber membrane and the precision of the

hollow fibers cartridge assembly process results in hollow fiber cartridges that perform

consistently and show high lot-to-lot consistency (Table 5)

INHERENTLY SUPERIOR CHARACTERISTICS

WaterSep Green Line hollow fiber cartridges are characterized by:

Antifouling modified glycerin free polyethersulfone (m-PES) composition for

regulatory satisfaction

Void-free structure for sustained reliable performance

Optimization for specific applications

Glycerin free membrane for very low extractables level.

Ready to use with no pre-rinse required. Just a low volume buffer equilibration.

PROVEN PERFORMANCE ADVANTAGES

Low fouling and anti-dead spot design

High flow rate and total capacity

High batch-to-batch consistency

Low cost of ownership and better economics due to no pre-treatment or post cleaning

Product Information


Table 5. Air flow specifications for WaterSep Green single use hollow fiber cartridges

Product Name Membrane NMWC

Air Flow per Cartridge ml/min/7 barg

(ml/min/10 psig)


1, 3, 5, 10, 30, 50, 100, 300,

500, 750 KD

< 0.2

MiniDiscover24 Green < 0.4

Discover12 Green < 0.6


Explorer12 Green < 1.6



Investigator12 Green < 14



BioProducer12 Green < 135



Maximizer24 Green < 540


Product Name Membrane

Air Flow per Cartridge ml/min/0.35 barg (ml/min/5 psig)


0.1, 0.2, 0.45, 0.65 µm

< 0.2

MiniDiscover24 Green < 0.4














Product Information


STORAGE OF NEW AND USED CARTRIDGES

NEW CARTRIDGES

Store new hollow fiber cartridges unopened in their original packaging in a protected

location and out of direct sunlight. The temperature of the storage area should remain

between 4°C to 37°C (39°F to 99°F). Under these conditions, a new cartridge can be stored

for 24 months without influencing performance or leading to product alterations.

PREPARING YOUR CARTRIDGE FOR USE

The WaterSep Green Line single use hollow fiber cartridges require a minimum of pre-

conditioning, prior to use.

Extractables are approximately 80X less than a conventional glycerin conditioned hollow

fiber or cassette product. Prior to use there is no need for any extensive rinsing of the

hollow fiber membrane with WFI or purified water. A quick buffer equilibration is all that

is needed. This is a significant saving of WFI/purified water, cleaning chemicals and

valuable facility time.

Because the Green Line hollow fiber cartridges are all single use there is no need for

developing cleaning regimen or cleaning validation documentation.

WaterSep recommends integrity testing of the hollow fiber cartridges pre – and post use.

The preferred method is a pressure hold test at either 10 or 5 psi, (see ….

Validation Information


CHAPTER 3—VALIDATION

INFORMATION

The various product lines of WaterSep hollow fiber cartridges are constructed of the same

materials and use identical design elements and fluid path length. Hence hollow fiber

cartridge performance is scalable, and validating a cartridge at one size normally suffices

for validating cartridges of other sizes.

MEMBRANE WATER FLUX

A common way to measure clean membrane performance and determine the level

membrane recovery after cleaning is to measure the water permeability of the membrane

under controlled conditions. [However, membrane water flux does not necessarily have a

direct correlation to process flux in a biological fluid stream.]

The water permeability of WaterSep membranes, normalized to water viscosity at the

testing temperature, are provided in Table 6.

Table 6. Water permeability of WaterSep membranes

Ultrafiltration Membrane

NMWL Normalized Water Permeability (LMH/psi) 1

3 K 0.1–1

5 K 0.5–3

10 K 7–13

30 K 12–30

50 K 15–40

100 K 18–40

300 K 20–50

500 K 20–50

750 K 25–60

Microfiltration Membrane

0.1 um > 30

0.2 um > 100

0.45 um > 200

1 These numbers are based on Explorer-size quality control cartridges.



MEMBRANE SOLUTE REJECTION

WaterSep’s hollow fiber membranes have very well defined and narrow

retention/passage specifications. Every manufacturing batch is tested for quality

with known solutes and must conform to highly-defined limits for rejection and passage.

Each membrane cut-off is tested with two to three solutes (membrane markers).

Table 7 lists the retention passage data for different membrane cut-offs.

A narrow pore size distribution, guarantees consistent membranes batches over time

and minimizes membrane batch-to-batch variations. Note that there are no overlap

in retention/passage specifications, for various cut-offs/pore sizes.

If a membrane does not have a clear cut-off point between particle retention and

passage, the membrane may pass your product of interest or provide low product yield.

There will be also significant performance variations between membrane batches (wide

lot-to-lot variations).

By testing the retention and passage of known solutes through the membrane under

controlled conditions, and measuring the concentration of the solute in the retentate

and permeate in comparison to the concentration of the original sample, you can

determine the performance of the membrane.

Table 7. Results of solute passage through WaterSep membrane (% P – percent passage)

NMWL

PVP-K15

PVP-K30

PVP-K60S PVP-K90

Blue Dextran Bubble Point

MW = 9700

MW = 66,800

MW = 396,000

MW = 1,570,000

MW = 2,000,000 IPA H2O

%P %P %P %P %P psig

3 K < 20

5 K 20–40

10 K 40–75 < 20

20 K > 70 < 25

30 K > 85 20–40

50 K > 90 40–70 < 10

100 K

> 70 < 20

300 K

20–35 < 20

500 K

35–65 < 30

750 K

> 65 25–50 < 10 > 60

0.1 um

> 50 10–50 35–60 60–100

0.2 um

> 50 22–35 45–60

0.45 um

> 75 10–22 30–45



CARTRIDGE INTEGRITY TEST

NON-DESTRUCTIVE TESTING

Non-destructive integrity testing can be completed on hollow fiber cartridges pre-

and post-operation in order to prevent process failure and product loss. Detecting a failed

hollow fiber cartridge can eliminate process delays and allow for rapid re-processing

of the batch. There are two types of non-destructive testing: the bubble point test

and the diffusion test.

WaterSep recommends using bubble point test for both UF and MF membranes.

(1K – 750K and 0.1 um – 0.65 um)

Pressure hold, forward flow, and pressure decay are variations of the diffusion test.

INTEGRITY TEST - PRESSURE HOLD TEST

The pressure hold test, also known as pressure decay or pressure drop test, is an

alternative to the diffusion test and is based on similar principles. In the pressure hold

test, an accurate pressure gauge is used to monitor upstream pressure changes due to gas

diffusion through the wetted hollow fiber membrane.

The pressure hold value is dependent on the diffusional flow and upstream volume. It can

be calculated using the following equation:

Where:

D = diffusion rate (cc/min)

T = time (minutes)

Pa = atmosphere pressure (1 Atm. or 14.7 psi)

Vh = upstream volume of apparatus (cc)

DP = pressure drop (bar or psi)



PROCEDURE

1. Circulate water through the hollow fiber cartridge for 10 minutes. Ensure fluid

flows from both permeate ports.

2. Attach an external pressure source to the feed/retentate side of the hollow fiber

system.

3. Close the permeate side and open feed and retentate ports.

4. Pressurize the system slowly to the specified pressure. 10 psi for UF membranes,

5 psi for MF membranes.

5. Drain the system of any liquid upstream.

6. Close the retentate valve

7. Open the permeate valve.

8. Let the system equilibrate for 5 minutes, while any remaining liquid will pass

through the HF cartridge.

9. Close the feed port, and monitor any pressure decay in the system.

10. The system/cartridge is integral if the pressure gauge reads a positive upstream

pressure after 1 minute.

Average Integrity Test Results Delta P/min @ 0.7 barg (10 psig)

Membrane NMWC (KD) Batch 1 Batch 2 Batch 3

1



REFERENCE VALUES FOR WATERSEP FACTORY MEMBRANE DIFFUSION

TEST

Table 8. Results of membrane integrity tests for Investigator12 Cartridges

CARTRIDGE WATER FLUX

Cartridge water flux for a specific cartridge and molecular weight cut off can provide

valuable information when it is compared to membrane water flux data. The correlation

factor for water flux between a membrane and a specific hollow fiber cartridge can also

be used to correlate water flux for other sizes of hollow fiber cartridges.

A cartridge water flux study was conducted on membranes with different molecular

weight cut off values. Included in the study were multiple cartridges for each molecular

weight cut-off. The goal was to determine the average water flux values for the cartridge.

Average Airflow Integrity Test Results cc/min/ft2 @ 0.7 barg (10 psig)

Membrane NMWC (KD) Batch 1 Batch 2 Batch 3

1

10

10

10

3

5

10

30

50

100

300

500

750

Average Airflow Integrity Test Results ml/min/ft2 at 0.35 barg (5 psig)

Membrane NMWC (µm) Batch 1 Batch 2 Batch3

0.1

10

10

10

0.2

0.45

0.45



Table 9 shows the average water flux for Investigator12 cartridges with different

molecular weight cut off values (membrane pore size). All the cartridges had 1,300-cm2

of membrane surface area.

The cartridge water flux results are normalized to surface area and reported as LMH

(liter/m²/hour)/psig (

Figure 5, Figure 6, and Figure 7).

Table 9. Water flux results for an Investigator12 hollow fiber cartridges

Membrane Type

Average Water Flux LMH/psi at .68 barg (10 psig) of TMP

m-PES 30K MWCO 17.4

m-PES 300K MWCO 36.3

M-PES 0.2 um 333.7

Figure 5. Clean water flux for an Investigator12 Green, 30K MWCO, 1.0 mm ID



Figure 6. Clean Water flux for an Investigator12 Green, 300K MWCO, 1.0 mm ID

Figure 7. Clean Water flux for an Investigator12 Green, 0.2 um, 1.0 mm ID



CARTRIDGE CROSSFLOW

The crossflow rate is a critical process parameter for crossflow filtration processes. An

optimized crossflow rate minimizes gel layer formation on the membrane surface, leading

to optimum process flux and maximum transmission of ions and low molecular weight

substances. Optimized crossflow, in combination with low transmembrane pressure

(TMP), is particularly important for cell clarification processes where transmission of

proteins, antibodies, and other high-molecular-weight target products is crucial.

Insufficient crossflow can lead to increased gel layer formation. The result is a decrease

in process flux and an increase in the rejection of low molecular solutes, both of which

lower yield.

The crossflow rate of a hollow fiber cartridge is proportional to the differential pressure

(delta P) along the cartridge (i.e. the pressure difference between the feed and retentate

port). The optimum crossflow rate is a function of the number of fibers in a hollow fiber

cartridge, the inside diameter of the fibers, and the fluid stream characteristics.

While the optimum cross flow rate for a family of hollow fiber cartridges does not vary

with path length, the delta P needed to achieve the optimum cross flow rate can vary.

For example, the optimum crossflow for a BioProducer12/24/41 is the same, although

the delta P to achieve that flow rate varies as a function of the path length of the hollow

fiber cartridge.

WaterSep hollow fiber cartridges are designed for intra-cartridge consistency for delta P

and the crossflow rate, and this design feature ensure consistent performance. Intra-

cartridge inconsistency between delta P and crossflow results in process variances and

potential product yield decay.

Figure 8 and Figure 9 show the measured crossflow vs. delta P forBioProducer12/24/41

Green and Investigator12/24/41 Green HF cartridges.



Figure 8. Delta P versus crossflow with water for Producer12/24/41 Green cartridges

Delta P vs. Cross Flow for Producer24

0

0.2

0.4

0.6

0.8

1

1.2

1.4

1.6

1.8

2

20 25 35 40 50

Delta P (psi)

Delta P

Cross FlowL/min


0

0.1

0.2

0.3

0.4

0.5

0.6

0.7

0.8

0.9

1

20 25 35 40 50

Delta P (psi)

Delta P

Cross FlowL/min


0

0.5

1

1.5

2

2.5

3

3.5

20 25 35 40 50

Delta P (psi)

Delta P

Cross FlowL/min



Figure 9. Delta P versus crossflow with water for Investigator12/24/41 Green cartridges

Delta P vs. Cross Flow for Investigator12

0

0.1

0.2

0.3

0.4

0.5

0.6

0.7

0.8

0.9

1

2 2.5 3.5 4 5

Delta P (psi)

Delta P

Cross FlowL/min


0

0.2

0.4

0.6

0.8

1

1.2

1.4

1.6

1.8

2

2 2.5 3.5 4 5

Delta P (psi)

Delta P

Cross FlowL/min


0

0.5

1

1.5

2

2.5

3

3.5

2 2.5 3.5 4 5

Delta P (psi)

Delta P

Cross FlowL/min



CARTRIDGE CHEMICAL COMPATIBILITY STUDY

GENERAL CONSIDERATIONS

Certain chemicals can damage and degrade the membrane structure, or ultimately

dissolve the membrane polymer. It is therefore important to be aware of the

membrane’s chemical compatibility and use compatible process and cleaning chemicals.

Under normal operating conditions, WaterSep cartridges are resistant to commonly-used

aqueous solutions within ph 1–14, bio-chemicals and most solvents, with the exception of

aromatic compounds. Certain operating conditions and elevated temperature, can affect

the membrane’s compatibility. WaterSep advises you to complete your own chemical

compatibility studies under you specific operating conditions and temperatures

(Table 10).

Table 10. Chemical compatibility list for WaterSep cartridges (R = recommended,

L = limited exposure, NR = not recommended, U = unknown)

Chemical Compatibility

Acetic acid (diluted-5%) R

Acetic acid (med conc-25%) L

Acetic acid (glacial) NR

Acetone NR

Acetonitrile NR

Alconox (1%) R

Ammonium hydroxide R

Amyl acetate NR

Amyl alcohol L

Aniline NR

Benzene NR

Benzyl alcohol NR

Boric acid R

Brine R

Butyl acetate NR

Butyl alcohol R

Butylaldehyde NR

Carbon tetrachloride NR

Chloroacetic acid NR

Chloroform NR

Chromic acid NR

Citric acid (2%) R

Cresol NR

Cyclohexane L


Cyclohexanone NR

Diacetone alcohol NR

Dichloromethane L

Dimethyl formamide NR

Dimethylsulfoxide (50%) L

1,4 Dioxane L

Ethers NR

Ethyl acetate NR

Ethyl Alcohol R

Ethyl alcohol (15%) R

Ethyl alcohol (95%) L

Ethylene dichloride NR

Ethylene glycol R

Ethylene oxide R

Formaldehyde (2%) R

Formaldehyde (30%) R

Formic acid (25%) R

Formic Acid (50%) R

Freon® R

Gasoline L

Glycerine / Glycerol R

Hexane R

Hexanol R

Hydrochloric acid (diluted- R

http://www.spectrumlabs.com/generic/tms.html?LiFrom=%2Fdialysis%2FCompatibility.html;FrName=Chemical+Compatibility;rtm=Freon;#FREON




5%)

Hydrochloric acid (conc-25%)

R

Hydrochloric acid (conc-37%)

R

Hydrofluoric acid (25%) L

Hydrogen peroxide (30%) L

Iodine solutions NR

Isobutyl alcohol R

Isopropanol R

Isopropyl acetate NR

Isopropyl alcohol / Isopropanol

R

Isopropyl ether R

Lactic acid R

Methyl acetate NR

Methyl alcohol R

Methyl alcohol (98%) L

Methyl cellosolve R

Methyl chloride NR

Methyl ethyl ketone NR

Methyl formate NR

Methyl isobutyl ketone NR

Methylene chloride NR

N-methyl-2-pyrrolidone NR

Mineral spirits R

Monochlorobenzene NR

NALCON 7647 (



CARTRIDGE RINSING AND BUFFER EQULIBRATION STUDY

A total of 650 ml of purified water was first flushed through a BioProducer12, hollow fiber

cartridge (1.25 m2, 13.5 ft2 membrane surface area) at a delta P of 0.2 barg (3 psig) from

feed to retentate port with the permeate port closed.

The retentate port was then closed. And an additional 650 ml of purified water was

flushed through the permeate ports at 0.33 barg (5psig) of TMP. Throughout the rinsing,

water samples were collected and analyzed for extractables in PPM.

After rinsing a total of 650 ml saline buffer ml was first flushed through the cartridge at a

delta P of 0.2 barg (3 psig) through the retentate port to drain with the permeate port

closed.

The retentate port was then closed, and an additional 650 ml of saline buffer were flushed

through the permeate port at 0.33 barg (5 psig) of TMP, to equilibrate the Bioproducer12.

Results shows that a minimum of 0.65 liters of purified water (0.5 L/m2 of membrane) was

required for rinsing the hollow fiber lab cartridge (Figure 10). This rinsing volume should

be proportionally adjusted for larger/smaller cartridges. The minimum rinsing volume—

0.65 liters for1.25 m2 (13.5 ft2) cartridges—should be proportionally increased when

larger cartridges with more membrane surface areas are used ( ≈ 0.5 L/m2 of membrane).

Figure 100. Results of Extractables Level vs. Volumetric Throughput of Purified Water

(Liters per m2)



HOLLOW FIBER CARTRIDGE SCALABILITY STUDIES

A key benefit of WaterSep hollow fiber cartridges is the capability to linearly and

consistently scale your process up or down from lab-scale to pilot-scale to production-

scale. Linear and consistent scaling reduces the costs and complexity of product

development and troubleshooting. The similarity in flow geometry between the different

product groups enables this capability.

WaterSep performed these two studies to demonstrate the scalability characteristics of

our cartridges:

1. An E. Coli lysate clarification study with Explorer12 and Explorer24 cartridges

2. A protein concentration study with Explorer12 and Investigator12 cartridges

E.COLI STUDY WITH EXPLORER12 AND EXPLORER24 CARTRIDGES

This study demonstrates cartridge scalability and provides data to assist with scalability.

The E. Coli lysate clarification study was conducted using Explorer12 and Explorer24

cartridges. The results of the study are illustrated in Figure 11, Figure 12, and Table 11.

CONCLUSIONS

The results indicate that TMP and flux data for production Explorer12 and Explorer24

have a scalability factor close to 1.0 in this application.

Figure 111. Pressure and permeate flux profiles for the Explorer12 cartridge



Figure 122. Pressure and permeate flux profiles for the Explorer24 cartridge

Table 11. Results from the E. Coli lysate clarification scalability study

Process Parameter

Explorer12 NMWL = 750 KD

Fiber Diameter = 1 mm

Explorer24 NMWL = 750 KD

Fiber Diameter = 1 mm

Fluid path length (inches) 12 24

Membrane surface area (cm2) 155 321

P feed (psi) 6–8 9.5–11.5

P retentate (psi) 1–2 1

TMP (psi) 3–4 2–4

Flux clarification (LMH) 18–20 21

Flux diafiltration (LMH) 24–31 24–33



PROTEIN CONCENTRATION STUDY WITH EXPLORER12

AND INVESTIGATOR12 CARTRIDGES

This study demonstrates cartridge scalability and provides data to assist with scalability.

A protein concentration study was conducted using Explorer12 and Investigator12

cartridges. The results of the study are illustrated in Figure 13.

Figure 133. Optimization results of scalability study using Explorer12 and Investigator12

cartridges

CONCLUSIONS

The Flux vs. TMP curves for an Explorer12 and an Investigator12 are almost identical and

suggest that the scalability factor is close to 1.0 in this application.

Product Safety


CHAPTER 4—PRODUCT

SAFETY

BIOCOMPATIBILITY STUDIES

WaterSep submitted samples of hollow fiber cartridges to Toxikon for biocompatibility

analyses. The following summarizes the results of the testing.

USP Class VI Test – GLP Compliance

Study Summary: “Therefore, the test article, WaterSep hollow fiber cartridge, meets the

requirements of USP guidelines, for Class VI Plastics – 70°C.”

USP Physicochemical Test for Plastics – GLP Compliance

Study Summary: “The purified water extract of the test article, WaterSep hollow fiber

cartridge, meets the test criteria described in the USP Physicochemical Test for Plastics

guidelines.”

You can review the Toxikon reports cited above by clicking on the links in Table 12.

Quality Assurance Information


CHAPTER 5—QUALITY

ASSURANCE INFORMATION

CERTIFICATE OF COMPLIANCE

WaterSep supplies a Certificate of Analysis with each cartridge. The Certificate of Analysis

verifies that your new cartridge was tested and complied with WaterSep’s quality

assurance standards (Figure 14).

Figure 144. Example of a cartridge certificate of compliance

Appendices


APPENDICES

APPENDIX I—ADDITIONAL DOCUMENTATION

You can learn more about WaterSep’s standard technology and obtain additional

information such as operating procedures and product sheets by visiting the WaterSep

website or by clicking a link below (Table 12).

Table 12. Helpful information available at www.WaterSep.net

Standard Operating Procedures

Hollow Fiber Cleaning and Storage Procedure

Preconditioning for Use Procedure

Product Sheets

MiniDiscover 12

MiniDiscover 24

Discover 12

Discover 24

Explorer12

Explorer24

Explorer 41

Investigator 12

Investigator 24

Investigator 41

BioProducer 12

BioProducer 24

BioProducer 41

http://www.watersep.net/http://watersep.net/shop/validation/HF%20SOP%20-%20CIP%20&%20Storage%20Procedure.pdfhttp://watersep.net/shop/validation/HF%20SOP%20-%20Preconditioning.pdfhttp://watersep.net/shop/pdf/WaterSep_MiniDiscover12.pdfhttp://watersep.net/shop/pdf/WaterSep_MiniDiscover24.pdfhttp://watersep.net/shop/pdf/WaterSep_Discover12.pdfhttp://watersep.net/shop/pdf/WaterSep_Discover24.pdfhttp://watersep.net/shop/pdf/WaterSep_Explorer12.pdfhttp://watersep.net/shop/pdf/WaterSep_Explorer24.pdfhttp://watersep.net/shop/pdf/WaterSep_Explorer41.pdfhttp://watersep.net/shop/pdf/WaterSep_Investigator12.pdfhttp://watersep.net/shop/pdf/WaterSep_Investigator24.pdfhttp://watersep.net/shop/pdf/WaterSep_Investigator41.pdfhttp://watersep.net/shop/pdf/WaterSep_BioProducer12.pdfhttp://watersep.net/shop/pdf/WaterSep_BioProducer24.pdfhttp://watersep.net/shop/pdf/WaterSep_BioProducer41.pdf

Appendices


APPENDIX II—TEST PROCEDURES AND REPORTS

You can obtain the reports cited in this validation guide from the WaterSep website by

clicking on the links below (Table 13).

Table 13. Test reports available from www.WaterSep.net

Title of Report

WaterSep Membrane and Cartridge Regulatory and Bio-Compatibility Summary Rev. 0 - AH

Toxikon Final GLP Report: 09-0302-G1 Class VI Test – USP

Toxikon Final GLP Report: 09-0302-G2 Physicochemical Test for Plastics – USP

http://www.watersep.net/http://watersep.net/shop/validation/BioCompatibility%20Summary.pdfhttp://watersep.net/shop/validation/WaterSep%20USP%20Class%20VI%20results%2009-0302-G1.pdfhttp://watersep.net/shop/validation/WaterSep%20USP%20physicochemical%20test%20results%2009-0302-G2.pdf

Index


INDEX

INDEX

Appendices, 41

Applications, 9

Audience for this guide, 6

Biocompatibility, 39

Biocompatibility studies, 39

Cartridge

additional information, 41

biocompatibility, 39

certificate of compliance, 40

chemical compatibility study, 33

cross-flow, 30, 35

design, 10

information, 9

inlets and outlets, 10

materials of construction, 14

packaging, 14

quality assurance, 40

storage, 22

Test procedures and reports, 42

water flux, 27

Cartridge scalability study, 36

Cartridges

Protein concentration, 38

scalablity, 36

Certificate of compliance, 40

Chemical compatibility, 33

Contacting WaterSep, 7

Cross-flow rate, 30

Customer support, 7

Documentation

additional information, 41

E-mailing WaterSep, 7

Extractables, 39

GLP compliance, 39

Help, 7

List of figures, 5

List of tables, 5

Materials of construction, 14

Membrane solute rejection, 24

Membrane water flux, 23

Non-destructive testing, 25

Packaging, 14

Performance

advantages, 20

Performance characteristics, 20

Preconditioning, 22

Pressure and permeate flux profiles, 36

Pressure hold test, 25

Process validation, 7

Protein concentration, 38

Quality assurance, 40

Rinsing

pH and conductivity reduction, 35

Rinsing cartridges, 34

Scalability, 36

Specifications, 15

materialls of construction, 14

physical properties, 15

water flux, 23

Storage, 22

Support, 7

Test procedures and reports, 42

Test reports, 42

USP Class VI Test, 39

USP Physicochemical Test for Plastics, 39

Validation, 7

Validation guide purpose, 6

Validation information, 23

Void volume, 15

Water flux, 27

Water permeability, 23

Website, 7




USA

Telephone: 508-970-0089 x204

Fax: 508-970-0146

Email:[email protected]

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