Sartorius Vivaspin 500 Operating Instructions Manual

85030-511-58
Technical data and operating instructions
Vivaspin® 500 µl and 2 ml
Vivaspin® 500 and 2 10K devices for in vitro diagnostic use Vivaspin
®
500 and 2 3K, 5K, 30K, 50K, 100K, 300K, 1000K and 0.2 µm
devices for research use only; not for use in diagnostic procedures For French, Italian, and Spanish manuals, please go to
www.sartorius.com/en/product-family/product-family-detail/m-centrisart-i/
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Vivaspin® 500 µl and 2 ml – Introduction
Storage conditions|shelf life
Vivaspin
®
ultrafiltration spin columns should be stored at 15 – 30°C. The devices should be used before the expiry date printed on the box.
Introduction
Vivaspin
®
Concentrators are disposable ultrafiltration devices for the concentration of biological samples. Vivaspin
®
500 is suit­able for sample volumes of 100–500 µl and the Vivaspin
®
2 can handle samples up to
2 ml. Vivaspin
®
2 can effectively be used in either swing bucket or fixed angle rotor accepting 15 ml centrifuge tubes.
The patented vertical membrane design and thin channel filtration chamber (US 5,647,990) minimises membrane fouling and provides high speed concentrations, even with particle laden solutions.
Vivaspin
®
500 can be used in a benchtop fixed angle rotor, accepting 2.2 ml centrifuge tubes.
C
The Vivaspin® 500 & 2 product line includes 9 different cutoffs (Molecular Weight Cutoff, MWCO): – Vivaspin
®
500 & 2 3K device: 3,000 MWCO
– Vivaspin
®
500 & 2 5K device: 5,000 MWCO
– Vivaspin
®
500 & 2 10K device:
10,000 MWCO
– Vivaspin
®
500 & 2 30K device:
30,000 MWCO
– Vivaspin
®
500 & 2 50K device:
50,000 MWCO
– Vivaspin
®
500 & 2 100K device:
100,000 MWCO
– Vivaspin
®
500 & 2 100K device:
300,000 MWCO
– Vivaspin
®
500 & 2 100K device:
1000,000 MWCO
– Vivaspin
®
500 & 2 100K device: 0.2 µm
Vivaspin
®
500 & 2 10K filter devices are for in vitro diagnostic use and can be used to concentrate serum, urine, cerebrospinal fluid, and other body fluids prior to analysis. Vivaspin
®
500 & 2 3K, 5K, 30K, 50K, 100K, 300K, 1.000K and 0.2 µm filter devices are for research use only and not for use in diagnostic procedures. The Vivaspin
®
500 & 2 devices are supplied non-sterile and are for single use only.
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Vivaspin® 2
The Vivaspin
®
2 is specifically designed with low internal surface and membrane area in order to achieve superior recoveries from very dilute solutions.
Another feature of the Vivaspin
®
2 is the choice of directly pipetting the concentrate from the dead stop pocket built into the bottom of the concentrator, or alternatively reverse spinning the concentrate into the recovery cap.
Membrane Alternatives
In addition to the proven high flux polyethersulfone (PES) membrane range which is recommended with most solutions, Vivaspin
®
2 is additionally offered with
cellulose triacetate (CTA) and Hydrosart
®
.
CTA is particularly recommended when high recovery of the filtrate solution is of primary importance. Hydrosart
®
is a stabilised cellulose based membrane that has been optimised for the biotechnological industry. The Hydrosart
®
membrane is a stable polymer
that features a broad pH range. Hydrosart
®
is also extremely hydrophilic, making it non-protein binding, virtually non-foul, and has extremely high flux. Hydrosart
®
is available in 5k, 10k, and 30k molecular weight cutoffs.
Please note that membrane behaviour largely depends on the specific characteristics of the solution being processed. Sartorius Stedim Biotech recommends that users experiment with alternative membranes in seeking to optimise their process performance.
Equipment Required
1. Centrifuge with swing bucket of fixed angle (minimum 25°) rotor.
Device Carrier Required
Vivaspin
®
500 2.2 ml/11 mm d
Vivaspin
®
2 15 ml/17 mm d
2. Pipettes for sample delivery and removal. For maximum recovery a thin gel loader type is recommended.
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Operation
1. Select the most appropriate membrane for your sample. For maximum recovery select a MWCO at least 50% smaller than the molecular size of the species of interest.
2. Fill concentrator with up to maximum volumes shown in table 1. (Ensure lid 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 type and MWCO.
5. Once the desired concentration is achieved, (see tables 3a & 3b for guide to concentration times), remove assembly and recover sample from the bottom of the concentrate pocket with a pipette.
Removing the Vivaspin
®
2 body from
the filtrate tube
The sleeve (seen from the end) is oval in cross section. The tube is round in cross 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 shape – before removing it with a twisting action.
Press here
Press here
Tube Sleeve
Reverse spin with Vivaspin® 2
Depending on user preference and need for sample storage, the concentrate can be reverse spun into the concentrate recovery cap (when fixed angle rotors are used, angle concentrator so that the printed window faces upwards|outwards). In this procedure remove filtrate tube, invert the concentrator body, insert concentrate recovery cap into filtrate tube and then spin at up to 3,000 g for 2 minutes.
Desalting|Buffer Exchange
1. Concentrate sample to desired level.
2. Empty filtrate container.
3. Refill concentrator with an appropriate solvent.
4. Concentrate the sample again and repeat the process until the concentration of contaminating microsolute is sufficiently reduced. Typically 3 wash cycles will remove 99% of initial salt content.
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Vivaspin® 2 Reverse Spin
Filtrate and concentrate can be sealed for storage.
Concentrate recovery cap
Concentrate
Position with printed window upwards|outwards
Concentrator body
Filtrate tube
Filtrate
Equipment required Vivaspin® 500 Vivaspin® 2 Centrifuge
Rotor type Fixed angle Swing bucket or Fixed angle Minimum rotor angle 40° 25° Rotor cavity To fit 2.2 ml (11 mm) To fit 15 ml (17 mm)
conical bottom tubes conical bottom tubes
Concentrate recovery
Pipette type Fixed or variable volume Fixed or variable volume Recommended tip Thin gel loader type Thin gel loader type
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Technical Specifications
Table 1: Technical specifications
Vivaspin
®
500 Vivaspin® 2
Concentrator capacity
Swing bucket rotor do not use 3 ml Fixed angle rotor 500 µl 2 ml
Dimensions
Total length 50 mm 126 mm Width 11 mm 17 mm Active membrane area 0.5 cm
2
1.2 cm
2
Hold-up volume, <5 µl <10 µl membrane and support
Dead stop volume* 5 µl 8 µl
Materials of construction
Body Polycarbonate Polycarbonate Filtrate vessel Polypropylene Polycarbonate Membrane Polyethersulfone PES, CTA, HY
Table 2: Recommended Spin Speed (x g)
Device Vivaspin
®
500 Vivaspin® 2
Membrane Fixed angle Fixed angle Swing bucket
3–50,000 PES 12,000 8,000 4,000 >100,000 PES 12,000 8,000 4,000 5–20,000 CTA 8,000 4,000 Hydrosart
®
8,000 4,000
* Dead stop volume as designed in moulding tool. This volume may vary depending on
sample, sample concentration, operation temperature and centrifuge rotor.
Usage Tips
1. Flow Rate
Filtration rate is affected by several parame­ters, including MWCO, porosity, sample con­centration, viscosity, centrifugal force and temperature. Expect significantly longer spin times for starting solutions with over 5% sol­ids. When operating at 4°C, flow rates are approximately 1.5 times slower than at 25°C. Viscous solutions such as 50% glycerine will take up to 5 times longer to concentrate than samples in a predominantly buffer solution.
2. Pre-rinsing
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.
3. Sanitization of Polyethersulfone Membranes
Vivaspin
®
devices should not be autoclaved as high temperatures will substantially increase membrane MWCO. To sanitize, use a 70% ethanol solution or sanitizing gas mixture.
4. Chemical Compatibility
Vivaspin
®
concentrators are designed for use with biological fluids and aqueous solutions. For chemical compatibility details, refer to table 4.
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Performance Characteristics
Table 3a: Typical Performance Characteristics Vivaspin® 500
Time to concentrate up Concentrate to 30x [min.] at 20°C recovery %
Start volume 500 µl 500 µl
Aprotinin 0.25 mg/ml (6,500 MW) 3,000 MWCO PES 30 96% BSA 1.0 mg/ml (66,000 MW) 5,000 MWCO PES 15 96% 10,000 MWCO PES 5 96% 30,000 MWCO PES 5 95% IgG 0.25 mg/ml (160,000 MW) 30,000 MWCO PES 10 96% 50,000 MWCO PES 10 96% 100,000 MWCO PES 10 96%
Table 3b: Typical Performance Characteristics Vivaspin® 2
Time to concentrate up Concentrate to 30x [min.] at 20°C recovery %
Start volume 2 ml 2 ml
Insulin chain A 0.1 mg/ml (2,535 MW) 2,000 MWCO Hydrosart
®
35 95% Aprotinin 0.25 mg/ml (6,500 MW) 3,000 MWCO PES 50 96% BSA 1.0 mg/ml (66,000 MW) 5,000 MWCO PES 12 98% 5,000 MWCO CTA 50 96% 5,000 MWCO Hydrosart
®
22 98% 10,000 MWCO PES 8 98% 10,000 MWCO CTA 10 96% 10,000 MWCO Hydrosart
®
12 98% 20,000 MWCO CTA 5 96% 30,000 MWCO PES 8 97% 30,000 MWCO Hydrosart
®
5 97% IgG 0.25 mg/ml (160,000 MW) 20,000 MWCO CTA 6 97% 30,000 MWCO PES 10 96% 50,000 MWCO PES 10 96% 100,000 MWCO PES 8 95%
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Chemical Compatibility
Table 4: Chemical Compatibility (2hr contact time)
Solutions PES CTA HY Compatible pH range pH 1–9 pH 4–8 pH 1–9
Acetic Acid (25.0%) OK NO OK Acetone (10.0%) NO NO NO Acetonitrile (10.0%) NO NO NO Ammonium Hydroxide (5.0%) ? OK OK Ammonium Sulphate (saturated) OK ? ? Benzene (100%) NO NO NO n-Butanol (70%) ? NO ? Chloroform (1.0%) NO NO NO Dimethyl Formamide (10.0%) ? NO NO Dimethyl Sulfoxide (5.0%) OK NO NO Ethanol (70.0%) OK OK OK Ethyl Acetate (100%) NO NO NO Formaldehyde (30%) OK OK OK Formic Acid (5.0%) OK ? OK Glycerine (70%) OK OK OK Guanidine HCI (6 M) OK ? OK Hydrocarbons, aromatic NO NO NO Hydrocarbons, chlorinated NO NO NO Hydrochloric Acid (1 M) OK NO OK Imidazole (300 mM) OK NO ? Isopropanol (70%) OK OK OK Lactic Acid (5.0%) OK NO OK Mercaptoethanol (1.0 M) NO NO OK Methanol (60%) ? ? OK Nitric Acid (10.0%) OK NO NO
Solutions PES CTA HY Compatible pH range pH 1–9 pH 4–8 pH 1–9
Phenol (1.0%) ? ? NO Phosphate Buffer (1.0 M) OK OK OK Polyethylene Glycol (10%) OK ? ? Pyridine (100%) NO NO NO Sodium Carbonate (20%) OK NO ? Sodium Deoxycholate (5.0%) OK ? ? Sodium Dodecylsulfate (0.1 M) OK OK OK Sodium Hydroxide (2.5 M) NO NO NO Sodium Hypochlorite (200 ppm) OK NO NO Sodium Nitrate (1.0%) OK ? OK Sulfamic Acid (5.0%) OK NO ? Tetrahydrofuran (5.0%) NO NO NO Toluene (1.0%) NO NO NO Trifluoroacetic Acid (10%) OK NO OK Tween
®
* 20 (0.1%) OK OK OK
Triton
®
** X-100 (0.1%) OK OK OK
Urea (8 M) OK ? OK
OK = Acceptable ? = Questionable NO = Not recommended
* Triton
®
is a registered trademark of Union Carbide Corp.
** Tween
®
is a registered trademark of ICI Americas Inc.
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Ordering Information
Ordering Tips
– Choose a membrane pore size at least
50% smaller than the size of the molecule to be retained.
– Usually choose Polyethersulfone
membranes for fastest concentrations.
– Usually choose Cellulose Triacetate for
Protein Removal|Ultrafiltrate recovery.
– Usually choose Hydrosart
®
membranes
for highest recovery with Ig fractions.
Vivaspin® 500 Polyethersulfone Qty per box Prod. no.
3,000 MWCO 25 VS0191 3,000 MWCO 100 VS0192 5,000 MWCO 25 VS0111 5,000 MWCO 100 VS0112 10,000 MWCO 25 VS0101 10,000 MWCO 100 VS0102 30,000 MWCO 25 VS0121 30,000 MWCO 100 VS0122 50,000 MWCO 25 VS0131 50,000 MWCO 100 VS0132 100,000 MWCO 25 VS0141 100,000 MWCO 100 VS0142 300,000 MWCO 25 VS0151 300,000 MWCO 100 VS0152 1,000,000 MWCO 25 VS0161 1,000,000 MWCO 100 VS0162
0.2 µm 25 VS0171
0.2 µm 100 VS0172 Starter pack (5 of each 5 k, 10 k, 30 k, 50 k, 100 k) 25 VS01S1
Vivaspin® 2 Polyethersulfone Qty per box Prod. no.
3,000 MWCO 25 VS0291 3,000 MWCO 100 VS0292 5,000 MWCO 25 VS0211 5,000 MWCO 100 VS0212 10,000 MWCO 25 VS0201 10,000 MWCO 100 VS0202 30,000 MWCO 25 VS0221 30,000 MWCO 100 VS0222 50,000 MWCO 25 VS0231 50,000 MWCO 100 VS0232 100,000 MWCO 25 VS0241 100,000 MWCO 100 VS0242 300,000 MWCO 25 VS0251 300,000 MWCO 100 VS0252 1,000,000 MWCO 25 VS0261 1,000,000 MWCO 100 VS0262
0.2 µm 25 VS0271
0.2 µm 100 VS0272 Starter pack (5 of each 5 k, 10 k, 30 k, 50 k, 100 k) 25 VS02S1
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Vivaspin® 2 Cellulose triacetate Qty per box Prod. no.
5,000 MWCO 25 VS02U1 5,000 MWCO 100 VS02U2 10,000 MWCO 25 VS02V1 10,000 MWCO 100 VS02V2 20,000 MWCO 25 VS02X1 20,000 MWCO 100 VS02X2
Vivaspin
®
2 Hydrosart® Qty per box Prod. no.
2,000 MWCO 25 VS02H91 2,000 MWCO 100 VS02H92 5,000 MWCO 25 VS02H11 5,000 MWCO 100 VS02H12 10,000 MWCO 25 VS02H01 10,000 MWCO 100 VS02H02 30,000 MWCO 25 VS02H21 30,000 MWCO 100 VS02H22
In Vitro Diagnostic Product Labeling
The following tabe defines the symbols found on Vivaspin
®
500 & 2 10K device labels.
Symbol SymbolDefinition Definition
In vitro diagnostic medical device
Catalogue number
Do not reuse
Use by
Batch code
Date of manufacture
Manufacturer
Temperature limitation
Non-sterile product
CE conformity marking
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Made in UK Sartorius Stedim Lab Ltd Sperry Way Stonehouse Park Gloucestershire GL10 3UT, UK
www.sartorius-stedim.com
Copyright by Sartorius Lab Instruments GmbH & Co. KG, Goettingen, Germany. 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.
Status: September 2018, Sartorius Lab Instruments GmbH & Co. KG, Goettingen, Germany
Specifications subject to change without notice. Copyright Sartorius Stedim Biotech GmbH. Printed in the EU on paper bleached without chlorine. Publication No.: SLU6093-e180910 Ver. 09 | 2018
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