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Technical data and operating instructions
Vivaspin® 6 and 20 mlVivaspin® 6 and 20 10K devices for in vitro diagnostic use Vivaspin® Turbo 6 and 20 3K, 5K, 30K, 50K, 100K, 300K, 1000K and 0.2 µm devices for research use only; not for use in diagnostic procedures Vivaspin® 6 and 20 ml – Introduction
Storage conditions shelf life
Vivaspin® 6 & 20 10K filter devices are for in Vivaspin® 6 and 20 ultrafiltration spin vitro diagnostic use and can be used to con- columns should be stored at 15 – 30°C. The centrate serum, urine, cerebrospinal fluid, devices should be used before the expiry and other body fluids prior to analysis. date printed on the box.
Vivaspin® 6 & 20 3K, 5K, 30K, 50K, 100K, 300K, 1000K and 0.2 µm filter devices are for research use only and not for use in Vivaspin® concentrators are disposable ultra- diagnostic procedures. The Vivaspin® Turbo filtration devices for the concentration 4 and 15 devices are supplied non-sterile and or purification of biological samples. and are for single use only.
Vivaspin® 6 is suitable for sample volumes of 2–6 ml and the Vivaspin® 20 can handle samples up to 20 ml. Both products feature Vivaspin® concentrators can be used in twin vertical membranes for unparalleled swing bucket or fixed angle rotors accepting standard conical bottom tubes. In a single spin, solutions can be concentrated in excess Vivaspin® 20 purification alternatives include of 100 +. Samples are typically concentrated a diafiltration cup that allows one step in 10 to 30 minutes with macromolecular removal of salts and other contaminating recoveries in excess of 95%. micromolecules, and a gas pressure mode for increased flexibility and even faster The longitudinal membrane orientation and processing.
thin channel concentration chamber, provide optimum cross flow conditions even for The innovative design (US Patent No. particle laden solutions; the centrifugal force 5,647,990, second patent pending), ease pulling particles and solids away from the of use, speed and exceptional concentrate membrane to the bottom of the device. recoveries are the main features of the Macromolecules collect in an impermeable concentrators.
concentrate pocket integrally moulded below the membrane surface, thereby eliminating the risk of filtration to dryness. The Vivaspin® 6 and 20 includes 9 different cutoffs (Molecular Weight Cutoff, MWCO): When an appropriate centrifuge is unavail- – Vivaspin® 6 & 20 3K device: 3,000 MWCO able, or for single sample processing, – Vivaspin® 6 & 20 5K device: 5,000 MWCO Vivaspin® 20 can be filled with up to 15 ml – Vivaspin® 6 & 20 10K device: and pressurised for bench top concentration. For even faster processing, pressure can be – Vivaspin® 6 & 20 30K device: combined with centrifugal force. "Pressure- Fugation" is particularly suitable for viscous – Vivaspin® 6 & 20 50K device: samples such as serum, or when processing at low temperatures, and generaly when – Vivaspin® 6 & 20 100K device: minimum process time is essential.
– Vivaspin® 6 & 20 100K device: – Vivaspin® 6 & 20 100K device: – Vivaspin® 6 & 20 100K device: 0.2 µm B. For use with Pressure (Vivaspin® 20 only)
1. Vivaspin® 20 Pressure Head A. For use with centrifuge
(Product No. VCA200). 1. Centrifuge with swing bucket or fixed angle rotor (minimum 25°).
2. Charge Valve for Pressure Head (Product No. VCA005).
2. Pasteur or fixed volume pipettes for sample delivery and removal.
3. Air Pressure Controller (Product No. VCA002) or equivalent pressure regulator For use with Pressure and Centrifuge
1. All of the equipment shown in A. and B. Equipment required
Swing bucket Fixed angle Swing bucket Fixed angle Minimum rotor angle To fit 15 ml (17 mm) To fit 50 ml (30 mm) conical bottom tubes conical bottom tubes Optional pressure accessories for Vivaspin® 20
Air pressure controller (APC) complete with pressure gauge, Prod. no. VCA002
regulator, over-pressure safety valve, female conncetor and 1 m extension line (4 mm pneumatic tubing) with male and female connectors and 1 m of 6 mm inlet tubingCharge valve VS20 pressure head Concentrate recovery
Fixed or variable volume Fixed or variable volume Thin gel loader type Thin gel loader type Please note: Vivaspin® 20 (30 mm + 116 mm) is designed to fit into rotors that can accom- modate Falcon 50 ml conical bottom tubes, e.g. Beckman Allegra 25R with TS-5.1-500 swing-out rotor with BUC 5 buckets and 368327 adaptors; Beckman TA-10.250 25° fixed angle rotor with 356966 adaptors; Heraeus Multifuge 3 S-R with (Heraeus/Sor- vall) 75006445 swing out rotor with 75006441 buckets and adaptors for Falcon 50 ml conical bottom tubes.
In Centrifuge VS6 & 20
Removing the Vivaspin® 6 body from
1. Select the most appropriate membrane the filtrate tube
cut-off for your sample. For maximum The sleeve (seen from the end) is oval in recovery select a MWCO at least 50% smaller cross section. The tube is round in cross than the molecular size of the species of section to give a tight fit to the sleeve. To release the tube from the sleeve, you must pinch the tube – to press it into an oval 2. Fill concentrator with up to maximum shape – before removing it with a twisting volumes shown in table 1. (Ensure screw closure is fully seated) 3. Insert assembled concentrator into centrifuge (when fixed angle rotors are used, angle concentrator so that the printed window faces upwards outwards). 4. Centrifuge at speeds recommended in table 2, taking care not to exceed the maximum g force indicated by membrane 5. Once the desired concentration is achieved, (see table 3a and 3b for guide to concentration times), remove assembly and recover sample from the bottom of the concentrate pocket with a pipette. Using Gas Pressure (Vivaspin® 20 only)
1. Select appropriate membrane as above.
Using gas pressure with Vivaspin® 20
2. Fill concentrator (maximum 15 ml).
3. Fit Pressure Head (Prod. No. VCA200), and hand tighten to ensure an air tight seal.
4. Using Air Pressure Controller (APC), exchange the Female Coupling for the Charge Valve (Prod. No. VCA005), on the APC Extension line. Pressurise by pressing the Charge Valve into the inlet valve of the pressure head to achieve an air tight seal. 5. Either – stand assembly in a rack and allow to concentrate, Or – for faster processing, insert assembled, pressurised concentrator into centrifuge and spin (see table 1 for spin speeds). 6. Once the desired concentration has been reached, (see table 3b for guide to concentration times), remove assembly and de-pressurise by unscrewing cap.
7. Extract concentrate using a pasteur or fixed volume pipette.
Desalting Buffer Exchange
Vivaspin® 20 Diafiltration
1. Concentrate sample to desired level.
2. Empty filtrate container.
3. Refill concentrator with an appropriate Diafiltration cup is filled with buffer solution (Product No: VSA005) 4. Concentrate the sample again and repeat the process until the concentration of contaminating microsolute is sufficiently During concentration, reduced. Typically, 3 wash cycles will remove solvent in sample is 99% of initial salt content.
continuously replaced by fresh buffer solution.
Desalting with Vivaspin® 20
Salts and contaminants can be removed in Salts and contaminants a single step when using the special diafil- are progressively cleared tration cup available with the Vivaspin® 20. through membrane and This is due to the constant washing action into filtrate vessel (constant volume dialfiltration), of the buffer solution in the cup as it replaces solvent and salts passing through the ultrafiltration 1. Place 2 ml sample solution in the concentrator. (Larger volumes can be desalted by first concentrating down to 2 ml and decanting filtrate).
2. Empty filtrate container.
3. Insert diafiltration cup into concentrator and fill with 10 ml deionised water or buffer solution. Re-fit blue lid over the diafiltration 4. Repeat concentration process; over 98% of salts will be removed in this step.
5. Remove diafiltration cup and recover concentrated and purified sample.
Table 1: Technical Specifications
Swing bucket rotor
Fixed angle rotor With pressure head Dimensions
125 mm with pressure head Active membrane area Hold up volume of membrane Dead stop volume* Materials of construction
Concentrator cap Table 2: Recommended Spin Speed (xg)
3–50,000 MWCO PES >100,000 MWCO PES Vivaspin® 20
3–50,000 MWCO PES >100–300,000 MWCO PES * Dead stop volume as designed in moulding tool. This volume may vary depending on sample, sample concentration, operation temperature and centrifuge rotor.
1. Flow Rate
3. Sterilisation of Polyethersulfone
Filtration rate is affected by several parame- ters, including MWCO, porosity, sample con- Polyethersulfone membranes should not centration, viscosity, centrifugal force and be autoclaved as high temperatures will temperature. Expect significantly longer spin substantially increase membrane MWCO. times for starting solutions with over 5% To sterilise, use a 70% ethanol solution or solids. When operating at 4°C. flow rates are sterilising gas mixture.
approximately 1.5 times slower than at 25°C. Viscous solutions such as 50% glycerine will 4. Chemical Compatibility
take up to 5 times longer to concentrate Vivaspin® concentrators are designed for use than samples in a predominantly buffer with biological fluids and aqueous solutions. For chemical compatibility details, refer to Membranes fitted to Vivaspin® concentrators contain trace amounts of Glycerine and Sodium azide. Should these interfere with analysis they can be removed by rinsing fill volume of buffer solution or deionised water through the concentrator. Decant filtrate and concentrate before processing sample solution. If you do not want to use the pre-rinsed device immediately, store it in the refrigerator with buffer or water covering the membrane surface. Please do not allow the membrane to dry out.
Table 3a: Performance Characteristics Vivaspin® 6
Time to concentrate up to 30x [min.] at 20°C
and solute recovery %
Min. Rec. Min. Rec.
Cytochrome c 0.25 mg/ml (12,400 MW) 3,000 MWCO PES BSA 1.0 mg/ml (66,000 MW) 5,000 MWCO PES 10,000 MWCO PES 30,000 MWCO PES IgG 0.25 mg/ml (160,000 MW) 30,000 MWCO PES 50,000 MWCO PES 100,000 MWCO PES Latex beads 0.004% in DMEM +10% FCS (0.055 µm) 300,000 MWCO PES Latex beads 0.004% in DMEM +10% FCS (0.24 µm) Yeast 1.0 mg/ml (S. Cerevisiae) 0.2 µm PES Table 3b: Performance Characteristics Vivaspin® 20
Time to concentrate up to 30x [min.] at 20°C
and solute recovery %
Centrifuge Centrifuge Bench Cytochrome c 0.25 mg/ml (12,400 MW) 96% 60 96% – – BSA 1.0 mg/ml (66,000 MW) 5,000 23 99% 29 99% 50 98% 14 98% 16 98% 17 98% 32 97% 8 97% 13 98% 15 98% 32 97% 8 97% IgG 0.25 mg/ml (160,000 MW) 30,000 27 97% 20 95% 46 94% 13 97% 27 96% 22 95% 46 93% 13 96% 25 91% 20 90% 42 88% 12 94% Latex beads 0.004% in DMEM +10% FCS (0.055 µm) 300,000 20 99% 35 99% 10 99% – – Latex beads 0.004% in DMEM +10% FCS (0.24 µm) 1,000,000 MWCO PES Yeast 1.0 mg/ml (S. Cerevisiae) 15 95% 5 95% 20 95% 2 95% Table 4: Chemical Compatibility (2hr contact time)
Compatible pH range
Compatible pH range
Acetic Acid (25.0%) Lactic Acid (5.0%) Mercaptoethanol (10 mM) Acetonitrile (10.0%) Ammonium Hydroxide (5.0%) Nitric Acid (10.0%) Ammonium Sulphate (saturated) Phosphate Buffer (1.0 M) Polyethylene Glycol (10%) Chloroform (1.0%) Dimethyl Formamide (10.0%) Sodium Carbonate (20%) Dimethyl Sulfoxide (5.0%) Sodium Deoxycholate (5.0%) Sodium Dodecylsulfate (0.1 M) Ethyl Acetate (100%) Sodium Hydroxide Formaldehyde (30%) Sodium Hypochlorite (200 ppm) Formic Acid (5.0%) Sodium Nitrate (1.0%) Sulfamic Acid (5.0%) Guanidine HCI (6 M) Tetrahydrofuran (5.0%) Hydrocarbons, aromatic Hydrocarbons, chlorinated Trifluoroacetic Acid (10%) Hydrochloric Acid (1 M) Tween®* 20 (0.1%) Imidazole (500 mM) Triton®** X-100 (0.1%) Isopropanol (70%) OK = Acceptable ? = Questionable NO = Not recommended * Triton® is a registered trademark of Union Carbide Corp.
** Tween® is a registered trademark of ICI Americas Inc.
Vivaspin® 6 Polyethersulfone
Qty per box
Starter pack (5 of each 5 K, 10 K, 30 K, 50 K, 100 K) Vivaspin® 20 Polyethersulfone
Qty per box
(2 of each 5 K, 10 K, 30 K, 50 K, 100 K, 0.2 µm) Vivaspin® 20 accessories
Qty per box
Air pressure controller (APC) Charge valve for pressure head Diafiltration cups Female connector 4 mm OD pneumatic tube (3 m) Vivaspin® 20 pressure head In Vitro Diagnostic Product Labeling
The following tabe defines the symbols found on Vivaspin® 6 & 20 10K device labels.
In vitro diagnostic medical device Date of manufacture Temperature limitation Non-sterile product CE conformity marking Sartorius Stedim Lab Ltd.
Unit 6, Stonedale Road, Sartorius Lab Instruments GmbH & Co. KG, Goettingen, All rights reserved. No part of this publication may be reprinted or translated in any form or by any means without the prior written permission of Sartorius Lab Instruments GmbH & Co. KG.
The status of the information, specifications and illustrations in this manual is indicated by the date given below. Sartorius Lab Instruments GmbH & Co. KG reserves the right to make changes to the technology, features, specifications and design of the equipment without notice.
Sartorius Lab Instruments Goettingen, Germany Specifications subject to change without notice. Copyright Sartorius Stedim Biotech GmbH. Printed in the EU on paper bleached without chlorine.
Publication No.: SLU6092-e151109
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