Sartobind®

Membrane Adsorbers for Rapid Purification of Proteins

turning science into solutions

Sartobind – the pace maker in Membrane Adsorber technology…

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l Fig. 1: Cytochrome c is eluted from a Sartobind S 75 Membrane Adsorber

Unique microporous structureSartobind Membrane Adsorbers display a macroporous structure with a pore size of > 3 μm.

That’s orders of magnitudes larger thanconventional chromatographic gel matrices. Molecules are transported by convective flow to ligands.

Characteristics of Membrane Adsorbers (MA)– Ready-to-use units– Simple handling– Pore sizes > 3 and 0.45 μm– Negligible diffusion limitation– Low bed heights with 0.3 up to 16 mm– Scalable to process dimension with

4 mm Sartobind disposable capsules and 8 mm batch reusable | disposableSartobind

– Robust high performance separations– No bed cracking, channeling,

air entrapment– Flow rate of ion exchange membranes

5–30 membrane volumes per minutebelow 2 bar (30 psi, 0.2 MPa)

– Chemistries: strong and weak ionexchange, coupling, affinity and metalchelate ligands

Low unspecific adsorptionThe basis for all Sartobind membranes is astabilized reinforced cellulose. It is madefrom regenerated cellulose and during theproduction to Sartobind it runs through anumber of stabilization and grafting stepsuntil a chromatographic matrix is formedon the cellulose backbone. In principle anyligands known from conventional chro-matography can be bound covalently onthe matrix.

Speed upSartobind ready-to-use units are run at 5 to 30 membrane volumes per minute.This is at least one order of magnitudefaster than chromatographic columns. A typical speed up factor is about 25 mea -sured in direct comparison to conventionalcolumn technology (reference: Walter, J. K. in: Bioseparation and Bioprocessing,Strategies and Considerations for Advanced Economy in Downstream Pro-cessing of Biopharmaceutical Proteins. G. Subramanian (ed.) Wiley VCH, Vol. ll p.447–460 (1998). Flow rate does not affectbreak through performance (see fig. 2).

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Fig. 2: Break through performance of a Sartobind 35 ml cation exchange module at mono clonal antibodypurification with various flow rates. For other pub-lished data see also: H.L. Knudsen et al., Genentech Inc.,J. Chromatogr. A 907 (2001) 145–154 .

Sartobind membrane types– Sartobind S, Q, STIC PA and D ion

exchangers– Sartobind IDA (iminodiacetic acid)

metal chelate– Sartobind Aldehyde– Sartobind Protein A (recombinant)

Sartobind applicationsPurification and Concentration– Large proteins, blood factors, protein

conjugates, viruses, VLP, mAbs Contaminant removal – Host cell proteins, DNA, viruses, leached

ligands, endotoxins, aggregates

… for robust separations

Chemical compatibilityThe housing of Sartobind SingleSep capsules is polypropylene. The housing ofSartobind MA 15, 75 and 100 is poly sulfonewhich is stable to many standard solventsapplied in chromato graphy. Sartobind ion exchange membranes are compatiblewith alcohols such as ethanol, isopropanol,glycerol, and denaturing solvents such as8 M urea and 8 M guanidine HCl and canbe cleaned with 1 N sodium hydroxide.

Constant capacityThe robustness of Sartobind MembraneAdsorbers in ion exchange and affinitychromatography has been tested in consecutive runs of 1000 chromatographiccycles, (Fig. 3). Sartobind can be reusedmany times, and affinity membranes maintain their binding capacity over manycycles reflecting the chemical stability ofthe ligands.

Fig. 3: Purification cycles of bovine serum albuminon Sartobind Q 75 strong anion exchanger repeated 1000 times. Flow rate: 120 cm/h, cycletime: 10 min, equilibration buffer: 20 mM phos-phate buffer pH 7.0, sample: 5 ml bovine serumdiluted 1:20 with equilibratioin buffer, elution buffer: 20 mM phosphate buffer pH 7.0 + 1 N NaCl,regeneration after each 100 cycles with 1 N NaOH

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Fig. 4: There are no restrictions regarding ligands and applications.

Quality assuranceMembranes are tested for protein dynamicbinding capacity, charge density, thicknessand flow rate. Capsules are tested fordynamic binding capacity and flow rate.

Validated technologySartobind 4 mm and 8 mm capsules havemet the requirements of current USPPlastic Class V and VI, respectively. Valida-tion guides are available with detailedtechnical data and test results according toUSP. Additionally, extractables guide isavailable for modules with data of GC-MS,HPLC and FT-IR analysis from aqueous,ethanolic and sodium hydroxide extraction.

FDA approvedSartobind is the first Membrane Adsorberapplied in approved pharmaceutical production (approval of CAMPATH-1H®

by Food and Drug Administration, March 2001).

Validated technology

Formats from lab to process

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Centrifugal formatIf you don’t need flow control but you have to screen a large number of proteins in parallel, use the Vivapure® centrifugalunits.

Sartobind MA unitsYou may use Sartobind Membrane Adsorbers for any ion exchange or affinitychromato graphy which require high speedand simple operation. The small scale units for laboratory use is the MA 15 with0.8 mm bed height (3 membrane layers),MA 75 with 4 mm bed height (15 layers)and MA 100 with 1.4 mm (5 layers).These may be reused hundreds of times.

Fig. 10: Vivapure centrifugal units

Fig. 5: Sartobind MAs may be used by hand or with a chromatographic system via Luer adapters.

Protein AThe Sartobind Protein A is the ideal tool for ultra rapid quantification ofimmunoglobulins (IgG). You may achievemuch more sample throughput per ml thanwith any other Protein A matrix. You canuse the unit by a syringe and perform thequantification within 10 minutes. Theenclosed Luer adapters allow direct con-nection to a liquid chromatography system.

Fig. 7: Sartobind Protein A 2 ml runs at 10 ml/min with a dynamic binding of 10–15 mg IgG per unit

Fig. 6: Sartobind MA units for laboratory applications

Fig. 9: Sartobind pico

Sartobind pico 0.08 mlThis format keeps the bed height of larger4 mm capsules and allows for small scaletrials with minute amount of samples e.g.for virus removal studies.

Work horse Sartobind MA 75 for lab purification workThe Sartobind MA 75 format with 4 mmbed height is available with all membranetypes. It can be scaled straightly to the production format with the same bedheight. They are designed for highthroughput laboratory purification of biomolecules in the range of 5–50 mg.

8 strips in 96 well platesThis format is suitable for process developers in biopharma to screen a largenumber of buffer conditions in a short time.

Fig. 11: 8 strip | 96 well plates

Production: Sartobind Jumbo reusableand disposableThe Jumbo is a 5 liter membrane chromatography device for large scale capturing and impurity removal at highflow rates. The Sartobind nano 3 ml is the appropriate scale down model withsame bed height of 8 mm.

Fig. 8: Sartobind capsules with 8 mm bed height: for single-use or batch-usebind and elute or flow through applications

Void volume optimized constructionSartobind 8 mm capsules are designed toachieve smallest peak | pool volumes. That is the reason for sharp peak resolutionof this capsule family and the applicabilityfor bind and elute purification processes.

Fig. 12: Sartobind 150 ml capsule cross section displayssmall upstream and downstream channels

Sartobind 4 mm capsules for contaminant removal

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Downscale with Sartobind pico and nanoThe Sartobind pico is the smallest member of the 4 mm capsule family with 0.08 mlmembrane volume scalable up to thelargest capsules. It saves costs in sampleconsumption and virus spiking studies.Sartobind nano 1 ml is the smallestcylindric capsule with 4 mm bed, Sartobindpico displays a flat sheet design to obtainthe smallest scalable device.

Membrane ligands available with 4 mm capsules– Quaternary ammonium – anion

exchanger– Sartobind STIC primary amine – salt

tolerant anion – Sulfonic acid – cation exchanger

Efficient– Higher throughput for trace impurity

removal at 10 kg protein per liter membrane in flow through mode

High speed polishingSartobind 4 mm capsules are designed toremove charged contaminants from thera-peutic proteins at accelerated flow rates of10 to 30 membrane volumes per minute byion exchange membrane chromatography.The high throughput is a direct result ofnegligible mass transfer effects and is made possible by the >3 μm macroporousmembrane with 4 mm bed height.

Capsule designThe Sartobind 4 mm looks like a standard filter capsule except that theadsorbing membrane is rolled up on a core to form a cylinder. The flow is fromtop into the outside channel and then perpendicular through the membrane layers to the center of the cylinder andleaves the capsule at the outlet.

pico nano mini 5” 10” 20” 30” mega

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Fig. 13: Sartobind 4 mm capsule scale up from ml to 10000 liter scale: Sartobind pico 0.08 ml to Sartobind mega 1.62 l

Economical– No hardware investment & maintenance– No column packing, testing,

regeneration– No re-use validation– Less labor and smaller foot print– Buffer consumption may be decreased

95% compared to columns– Sartobind STIC PA reduces tank invest-

ments and buffer usage as cationexchange pools are cleared from contam-inants without dilution

Easy to use– Disposable– Simple and fast set up– Handling like a filter capsule– Less complex

Unique for large proteins, DNA and virusesSartobind membranes are unique for purification of large proteins and virusparticles. Conventional gel beads possess95% of the binding capacity within thebead and large particles cannot enter their small pores. In contrast Sartobindmembranes display huge capacities forlarge molecules. E.g. for DNA 10 times more dynamic capacity than conventionalbeads (reference: Sartobind ApplicationNote DNA removal) and for virus purifica-tions gels cannot compete at all. Sartobindmembranes have been successfully appliedfor the purification of alphaherpesvirus, adenovirus and adenoassociated virus. The Vivapure AdenoPACK adenoviruspurification and concentration kits offerresearchers a convenient way to clean up within 1–2 hours. The LentiSELECT kitreduces the purification time of lenti viruses up to 8+106 particles below 1 hour.Both systems are scalable to productionformat.

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A standard for virus purification

Fig. 14: Viruses can bind to the internal membrane structure

Fig. 15: Adeno PACK for convenient adenovirus purification and concentration

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Technical Data

To connect to achromatography system or Luersyringe

pico MA 15 MA 75 | 2ml MA 100 nanoApplication Screening, applica- Screening, applica- Screening, applica- Down-scale

tion optimization, tion optimization, tion optimization, for biopharma reusable reusable reusable production

Membrane material Stabilized reinforced cellulose

Adsorption area [cm2] 2.9 15 75 | 100 100 36 | 110

Number of layers 15 3 15 | 20 5 15 | 30

Bed height [mm] 0.4 0.8 4.0 1.4 4 | 8

Bed volume* [ml] 0.08 0.41 2.1 | 2 2.8 1 | 3

Membrane diameter [mm] 5 25 25 50 –

Housing material Polypropylene Polysulfone Polysulfone Polysulfone Polypropylene

Inlet connector Female Luer Female Luer Female Luer Female Luer Female Luer

Outlet connector Female Luer Male Luer Male Luer Male Luer Female Luer

Typical dynamic binding 2.3 / Q pico 10.5 / S 15 52.5 / S 75 70 / S 100 29 / Q nano 1 mlcapacity 10% binding 4.0 / PA pico 12 / Q 15 60 / Q 75 80 / Q 100 25 / S nano 1 mlcapacity* [mg/unit] 2.0 / S pico 60 / D 75 60 / PA nano 1 ml

7.5 / IDA 75 88 / Q nano 3 ml15 / Protein A 77 / S nano 3 ml

44 / Phenyl nano 3 ml

Recommended flow rate 0.8–2.4 10–30 10–30 20–50 10–30 | 5–15[ml/min]

Maximum pressure [MPa] 0.6 0.6 0.6 0.6 0.4

* Reference proteins ion exchange: S: lysozyme, Q, D, PA: BSA, IDA: 6 + His-tagged protein, epoxy. IgG, Protein A: IgG, Phenyl: IgG

Overview Membrane Types and Formats

Mem- Description Pore Capacity* Sartobind® Lab Scale Process Scalebrane size (mg/ml) A4 96 well MA 15, MA pico Capsule FT Capsule FT/B&E**Type (μm) sheet 19 μl 100 75 0.08 ml 4 mm 8 mmS Sulfonic acid >3 25 • (•) • • • • •

Q Quaternary ammonium >3 29 • (•) • • • • •

PA Primary amine >3 60 (•) • •

D Diethylamine >3 22 • •

IDA Iminodiacetic acid >3 3.6 • •

HIC Phenyl >3 44 (•) (•) •

Aldehyde Aldehyde 0.45 1 •

Protein A Recombinant Protein A 0.45 5–7.5 •

* Typical dynamic binding capacity 10%** Flow through | Bind and elute• available, (•) coming soon

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For centrifuge

Mini MaxiApplication Screening, application Screening, application

optimization, disposable optimization, disposable

Membrane ligands Q, D, S Q, S

Membrane diameter [mm] 6.4 22.5

Housing material Polypropylene Polypropylene

Minimum static binding H: 4 H: 48–64capacity* [mg/unit]

Maximum volume [ml] H: 0.4 H: 19(device inner volume)

Spin speed [g] H: 2000 H: 500

Storage before use at Room temperature Room temperature

M: medium, H: high* Reference proteins ion exchange: S: lysozyme, Q, D: BSA

For centrifuge | vacuum For use with a syringe or liquid chromatography system

96 well plates equipped with 8 strips Sartobind pico Application Screening for process development work Screening and scale up for trials

Membrane ligands S, Q, STIC PA and HIC Phenyl HIC Phenyl

Housing material Polypropylen Polypropylen

Number of layers 3 15

Bed height 0.8 mm 4 mm

Membrane volume 0.019 ml 0.08 ml

Membrane diameter 5.5 mm 5 mm

Maximum well volume [ml] 0.5 ml –

Collection plate volume [ml] 2 ml –

Upcoming products

Ordering Information

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Luer Lock units|sheets|kits

Order No. Sartobind MA Quantity

93IEXQ42GB-12--A Sartobind Q 15 4

93IEXS42GB-12--A Sartobind S 15 4

93IEXQ42DB-12--V Sartobind Q 75 2

93IEXS42DB-12--V Sartobind S 75 2

93IEXD42DB-12--V Sartobind D 75 2

93IEXQ42BC-12 Sartobind Q 100 1

93IEXS42BC-12 Sartobind S 100 1

93IDA426DB-12--V Sartobind IDA 75 2

93PRAP06HB-12--A Sartobind Protein A 2 ml 4

Order No. Sartobind A4 Sheet Quantity

94IEXS42-001 Sartobind S A4 Sheet 1

94IEXQ42-001 Sartobind Q A4 Sheet 1

94IEXD42-001 Sartobind D A4 Sheet 1

94IDA-42-001 Sartobind IDA A4 Sheet 1

94ALD-06-001 Sartobind Aldehyde A4 Sheet 1

Sartobind capsules 4 mm bed height

Order No. Sartobind Q Quantity

92IEXQ42DD-11--D Sartobind Q pico 0.08 ml 10

92IEXQ42DN-11 Sartobind Q SingleSep 1nano 1 ml

92IEXQ42DN-11--A Sartobind Q SingleSep 4nano 1 ml

92IEXQ42D4-OO--A Sartobind Q SingleSep 4mini capsule, hose barb

92IEXQ42D4-SS--A Sartobind Q SingleSep 4mini capsule, sanitary

92IEXQ42D9-OO--A Sartobind Q SingleSep 45” capsule, hose barb

92IEXQ42D9-SS--A Sartobind Q SingleSep 45” capsule, sanitary

92IEXQ42D1-SS Sartobind Q SingleSep 110”capsule

92IEXQ42D2-SS Sartobind Q SingleSep 120”capsule

92IEXQ42D3-SS Sartobind Q SingleSep 130”capsule

92IEXQ42DC3SS Sartobind Q SingleSep 1mega capsule

Order No. Sartobind S Quantity

92IEXS42DD-11--D Sartobind S pico 0.08 ml 10

92IEXS42DN-11 Sartobind S SingleSep 1nano 1 ml

92IEXS42DN-11--A Sartobind S SingleSep 4nano 1 ml

92IEXS42D4-OO--A Sartobind S SingleSep 4mini capsule, hose barb

92IEXS42D4-SS--A Sartobind S SingleSep 4mini capsule, sanitary

92IEXS42D9-OO--A Sartobind S SingleSep 45” capsule, hose barb

92IEXS42D9-SS--A Sartobind S SingleSep 45” capsule, sanitary

92IEXS42D1-SS Sartobind S SingleSep 110”capsule

92IEXS42D3-SS Sartobind S SingleSep 130"capsule

Order No. Sartobind STIC Quantity

92STPA42DD-11--D Sartobind STIC PA 10pico 0.08 ml

92STPA42DN-11--A Sartobind STIC PA 4nano 1 ml

92STPA42D9-FF--A Sartobind STIC PA 45” capsule

92STPA42D1-SS Sartobind STIC PA 110” capsule

92STPA42D3-SS Sartobind STIC PA 130" capsule

92STPA42DC3SS Sartobind STIC PA 1mega capsule

Sartobind capsules 8 mm bed height

Order No. Sartobind Q Quantity

96IEXQ42EUC11--A Sartobind Q nano 3 ml 4

96IEXQ42E9BFF Sartobind Q 150 ml 1

96IEXQ42E3ESS Sartobind Q Jumbo 5 l 1

Order No. Sartobind S Quantity

96IEXS42EUC11--A Sartobind S nano 3 ml 4

96IEXS42E9BFF Sartobind S 150 ml 1

96IEXS42E3ESS Sartobind S Jumbo 5 l 1

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Order No. Sartobind Phenyl Quantity

96HICP42EUC11--A Sartobind Phenyl nano 3 ml 4

96HICP42E9BFF Sartobind Phenyl 150 ml 1

96HICP42E3ESS Sartobind Phenyl Jumbo 5 l 1

Order No. Kits | Accessories Quantity

VS-AVPQ020 Vivapure AdenoPack 20

VS-AVPQ022 Vivapure AdenoPack 20 RT

VS-AVPQ101 Vivapure Adenopack 100

VS-AVPQ102 Vivapure Adeno PACK 100 RT*

VS-AVPQ501 Vivapure Adeno PACK 500

VS-AVPQ502 Vivapure AdenoPack 500 RT

VS-LVPQ040 LentiSELECT 40

90-KIT-01 Sartobind Demo Kit

1ZA---0004 Adapter UNF 10–32 1to Luer male, PEEK

1ZA---0005 Adapter UNF 10–32 1to Luer female, PEEK

17002---140 Pair of Luer adapters, 2black Tefzel M6 thread

16517----------E Syringe filter holders, 25 mm 12diameter, polycarbonate for filter or adsorber membrane

16214 Syringe filter holder 25 mm 1diameter, stainless steel for filter or adsorber membrane

9ZAIAM0001 Legs for Sartobind 3mega capsule (stainless steel)

9ZGL--0102 Trolley for Sartobind Jumbo 15 l capsule

* Vivapure Adeno PACK RT does not contain Benzonase®

Spin columns

Order No. Vivapure Mini Spin Spin columns|Columns Centrifuge tubes

VS-IX01SH24 Vivapure S Mini H 24 | 48

VS-IX01QH24 Vivapure Q Mini H 24 | 48

VS-IX01DH24 Vivapure D Mini H 24 | 48

Order No. Vivapure Maxi Spin Spin columns|Columns Centrifuge tubes

VS-IX20SH08 Vivapure S Maxi H 8 | 16

VS-IX20QH08 Vivapure Q Maxi H 8 | 16

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