Lab Ultrafiltration and Purification Products Catalog
Lab Ultrafiltration and Purification Products Catalog
Lab Ultrafiltration and Purification Products Catalog
and Purification
Products
Table of Contents
General Information
Ultrafiltration Applications 4
Lab Ultrafiltration Devices 5
Ultrafiltration Process Methods 5
Membrane Performance Characteristics 6
Membrane Selection Guide 7
---
Protein and Macromolecule Concentration 9
Centrifugal and Pressurized Ultrafiltration
---
Vivaspin® 500 10
Vivaspin® 2 12
Vivaspin® Filtrate 14
---
Vivaspin® Turbo 4 PES 16
Vivaspin® 6 18
Vivaspin® 15R 20
---
Vivaspin® Endotest 22
Vivaspin® Turbo 15 PES 24
Vivaspin® Turbo 15 RC 26
Vivaspin® 20 28
Vivaspin® 100 30
---
Vivaspin® Equipment and Accessories 32
-
Vivaflow® 50 34
Vivaflow® 50R 36
Vivaflow® 200 38
Vivaflow® Equipment and Accessories 40
2
DNA Concentration 45
Vivacon® 500 46
Vivacon® 2 49
Protein Purification 53
Vivaclear Centrifugal Filters 54
Vivapure® Ion Exchange Purification Products 56
---
Vivapure® Virus Purification and Concentration Kits 60
Adenovirus Purification 61
Vivapure® Adenopack 20 62
Vivapure® Adenopack 100 63
---
5Vivapure® Adenopack 500 65
Lentivirus Purification 66
Vivapure® Lentiselect 40 67
Vivapure® Lentiselect 500 68
Vivapure® Lentiselect 1000 69
Application Notes 71
1. Desalting and Buffer Exchange with Vivaspin® Centrifugal
Concentrators 72
2. Treatment of Vivaspin® Concentrators for Improved Recovery
of Low-Concentrated Protein Samples 75
3. Scouting Protein Purification Conditions Using Vivapure® Centrifugal
Ion Exchange Membrane Absorbers 78
4. Concentration and Purification of Viruses by using Ultrafiltration,
Incl. Coronavirus - a Short Review 83
5. Sartorius Ultrafiltration Products in the Preparation of Biological
Nanoparticles and Medical Nanocarriers 95
6. Vivaflow® and Vivaspin® Workflow in Protein Research Laboratories 101
3
Ultrafiltration Applications
-
than alternative processes. Ultrafiltration is a cost effective
method for separating samples into
Typical Applications size-graded components providing
Concentration | desalting of proteins, that the desired fractions have at
--
enzymes, DNA, monoclonal antibod- least a 10-fold difference in molecular
ies, immunoglobulins, extracellular weight. During filtration, the
vesicles, viruses and nanoparticles permeating solute remains at its initial
Forensic DNA sample concentration concentration whilst the retained
--
prior to sequencing reaction macromolecules will be enriched.
Solute fractionation Peptide fractionation in FASP
(filter-aided sample preparation) Solute Desalting or Purification
--
Free drug | hormone assays A solution may be purified from salts,
Removal of primers from PCR solvents and low molecular weight
amplified DNA materials by diafiltration. Multiple
--
Removal of labeled amino acids solvent exchanges will progressively
and nucleotides purify macromolecules from
HPLC sample preparation contaminating microsolutes, which are
--
Deproteinization of samples typically removed most efficiently by
Recovery of biomolecules from cell adding solvent to the sample at a rate
culture supernatants | lysates equal to the speed of filtration. This is
-
Solute desalting or purification Mammalian cell harvesting called continuous diafiltration, and it
Cell washing, virus purification, cell replaces time-intensive techniques
debris removal and depyrogenation such as dialysis.
Environmental sample clarification |
concentration
4 General Information
Lab Ultrafiltration Devices
Sartorius develops devices dedicated support tools to help users select the
to optimizing laboratory ultrafiltration optimum device and process for their
processes with minimal time sample type.
requirements while maximizing
Vivaspin® Turbo
1.5
0.5
Centrifugal
Pressure
(5 to 98 mL starting volumes)
Pressurized air or an inert gas provide
the vector for ultrafiltration. For
increased process speed, pressurized
devices can be placed on an orbital
shaker, where agitation impedes
macromolecules from polarizing on
the membrane surface. Vivaspin® 20
and 100 can be operated using gas
pressure.
General Information 5
Membrane Performance Characteristics
Hydrosart® Concentration
2 kDa MWCO Desalting
5 kDa MWCO Buffer exchange
10 kDa MWCO Fractionation
30 kDa MWCO Membrane evaluation for scale up
6 General Information
Membrane Selection Guide
The advanced designs and low surface of the sample container. Whilst
adsorption materials that characterize the relative adsorption will be
Sartorius ultrafilters, offer a unique proportionately less important than on
combination of faster processing the membrane, due to the higher total
speeds and highest target molecule surface area, this can be the major
recoveries. Providing that the source of yield loss.
appropriate sample capacity,
membrane material and MWCO are Process Optimization
selected, these devices will typically When the highest recoveries are
yield recoveries in excess of 90% when crucial, particularly when working with
--
the initial sample contains > 0.1 mg/mL solute quantities in the microgram
of the solute of interest. The majority of range, Sartorius recommends that
any loss occurs through non-specific users consider the following:
binding to the membrane surface and | Select the smallest device that suits
or the sample container polymer. the sample volume.
--
Take advantage of the extra speed
Adsorption to the Membrane of Sartorius products by refilling a
Depending on sample characteristics smaller device repeatedly.
relative to the membrane type used, Select the lowest MWCO membrane
solute adsorption on the membrane that suits the application.
-
surface is typically 2-10 μg/cm2. This can Reduce pressure or centrifugal
increase to 20-100 μg/cm2 when the force to approximately half of the
filtrate is of interest and the solute must recommended maximum.
pass through the whole internal Avoid over-concentration. The
-
structure of the membrane. Typically a smaller the final concentrate volume,
higher cut-off membrane will bind the more difficult it is to achieve
more than a low molecular weight complete recovery.
-
alternative. If feasible, after sample retrieval,
rinse the device with one or more
Adsorption to the Sample Container drops of buffer.
Although every effort is made to Pretreat the device overnight with a
minimize this phenomenon by the passivation solution such as 5% SDS,
selection of low binding materials and Tween 20 or Triton X-100, then rinse
tool production to optical standards, thoroughly before use.
some solute will bind to the internal
For highest recovery, select a membrane MWCO which is a maximum of one third to half the
molecular weight of the solute to be retained
General Information 7
8 Chapter Site Title
Protein and Macromolecule
Concentration
Table of Contents
Vivaspin® 500 10
Vivaspin® 2 12
Vivaspin® Filtrate 14
Vivaspin® 6 18
Vivaspin® 15R 20
Vivaspin® Endotest 22
Vivaspin® Turbo 15 RC 26
Vivaspin® 20 28
Vivaspin® 100 30
Vivaflow® 50 34
Vivaflow® 50R 36
Vivaflow® 200 38
9
Vivaspin® 500
Technical Specifications
Concentrator capacity
Dimensions
100
50 75
Length x diameter 50 x 11 mm
50
50
Dead-stop volume 5 µL
Materials of construction
Equipment Required
Centrifuge
Concentrate recovery
Time Recovery
Ordering Information
1.5
Technical Specifications
1.0
100K
1.0
MWCO
126
200
µL
Concentrator capacity
100
Dimensions
Dead-stop volume 8 μL
Materials of construction
Equipment Required
Centrifuge
Concentrate recovery
Recommended tip Thin gel loader type Thin gel loader type
2.0
mL
1.5
Time to concentrate up to 30×
at 20°C and solute recovery
1.5
1.0
100K
1.0
MWCO
avoids risk of 20
100
BSA 1.0 mg/mL (66 kDa)
5 kDa MWCO PES 12 min 98%
µL
200
MWCO
1.0
5 kDa MWCO HY 22 min 98%
10 kDa MWCO PES 8 min 98%
100K
1.0
1.5
Ordering Information
Vivaspin 2 CTA
®
Vivaspin 2 HY
®
---
ultrafiltration to separate proteins from
low molecular weight substances in Vivaspin® Filtrate is ideal for the
biological samples. following applications:
--
Drug binding studies
Vivaspin Filtratre® features a unique Isolation of metabolites from serum
design that enables ultrafiltration in Protein removal from blood samples
the direction opposite to centrifugal Cleaning of liposomes
force. This is so effective in preventing Virus removal
premature blockage of the filter that
even whole blood samples can be
deproteinized.
Technical Specifications
Concentrator capacity
Swing bucket rotor 2.5 mL
93
Fixed angle rotor 2.5 mL
Dimensions
Length x diameter 93 x 14 mm
Active membrane area 0.79 cm2
Hold-up volume, membrane < 5 μL
max. 2.5 mL
Materials of construction
14
Centrifuge tube Polystyrene (PS)
Filtrate tube Styrene Acrylonitrile (SAN)
Concentrator cap Polyethylene (PE)
Membrane Cellulose Triacetate (CTA)
Polyethersulfone (PES)
Equipment Required
Centrifuge
Concentrate recovery
Recommended tip Thin gel loader type Thin gel loader type
Ordering Information
Technical Specifications
Concentrator capacity
122
Swing bucket rotor 4 mL
Fixed angle rotor 4 mL
Dimensions
Length x diameter 122.5 x 17 mm
Active membrane area 3.2 cm2
Hold-up volume, membrane < 10 μL
Materials of construction
Body Styrene Butadiene Copolymer (SBC)
Filtrate vessel Polypropylene (PP)
Concentrator cap Polypropylene (PP)
Membrane Polyethersulfone (PES)
Equipment Required
Centrifuge
Concentrate recovery
Recommended tip Thin gel loader type Thin gel loader type
--
Here you can find instruc- Rotor Swing bucket Fixed angle (25°)
tions on how to use Vivaspin® Centrifugal force* 4,000 g 7,500 g
Turbo 4 PES for:
Start volume 4 mL 4 mL
Desalting and buffer
Time Recovery Time Recovery
exchange
--
Preparation of biological Cytochrome c (12.4 kDa)
nanoparticles and medical 3 kDa MWCO PES 60 min 98% 80 min 96%
5 kDa MWCO PES 40 min 95% 50 min 94%
nanocarriers
Concentration and Lysozyme (14.3 kDa)
3 kDa MWCO PES 65 min 95% 70 min 93%
-
purification of viruses 5 kDa MWCO PES 50 min 94% 60 min 92%
Urine protein concentra-
α-Chymotrypsin (25 kDa)
tion
10 kDa MWCO PES 10 min 95% 8 min 95%
Separation of proteins
BSA (66 kDa)
and metabolites for disease
10 kDa MWCO PES 10 min 98% 7 min 97%
detection 30 kDa MWCO PES 8 min 96% 6 min 97%
Ordering Information
Technical Specifications
5
3 Concentrator capacity
2
Swing bucket rotor 6 mL
1
0.5
Fixed angle rotor 6 mL
0.3
Dimensions
0.2
122 0.1
Materials of construction
Body Polycarbonate (PC)
17
Filtrate vessel Polycarbonate (PC)
Concentrator cap Polypropylene (PP)
Membrane Polyethersulfone (PES)
Equipment Required
Centrifuge
Concentrate recovery
Recommended tip Thin gel loader type Thin gel loader type
Start volume 6 mL 6 mL
Ordering Information
--
--
2 to 15 mL samples Ultimate recoveries (95 – 98%)
Vivaspin® 15R is designed for initial Extremely short concentration time
sample volumes up to 15 mL and (30-fold in 15 minutes)
features a modified regenerated Simple and convenient handling
-
cellulose membrane; Hydrosart®. Easy scale-up to 0.1 to 5 L with
This membrane is ideal where Vivaflow® 50R or 200 with
extremely high recovery with very low Hydrosart® membranes
adsorption is needed. An example of Very low hold-up volume (< 20 μL)
this application includes desalting and
concentration of immunoglobulin
fractions.
15
10
Technical Specifications
5
Concentrator capacity
1.0
0.5
0.3
0.15
Dimensions
Materials of construction
Equipment Required
Centrifuge
Concentrate recovery
Recommended tip Thin gel loader type Thin gel loader type
0.3
0.5
0.75
1.0
10
0.5
0.3
1.0
10
10
15
15
5
0.75
0.15
0.5
0.3
1.0
10
1.0
0.5
0.3
0.15
15
α-chymotrypsin 0.25 mg/mL* (25 kDa)
10 5 kDa MWCO 50 min 98% 45 min 98%
10 kDa MWCO 25 min 98% 18 min 98%
5
1.0
0.75
0.5
0.15
Ordering Information
Vivaspin® 15R HY 12 pc 48 pc
15
10
Technical Specifications
5
Concentrator capacity
1.0
0.5
0.3
0.15
Dimensions
Materials of construction
Equipment Required
Centrifuge
Sample Mixing
Concentrate recovery
Recommended tip Thin gel loader type Thin gel loader type
0.3
0.5
0.75
1.0
10
15
0.5
0.3
1.0
10
10
15
15
5
0.75
0.15
0.5
0.3
1.0
15
10
1.0
0.75
0.5
0.3
0.15
Spin
15
10
1.0
0.75
0.5
0.3
0.15
Recover
1.5
Concentrator capacity
Dimensions
Materials of construction
Equipment Required
Centrifuge
Concentrate recovery
Recommended tip Thin gel loader type Thin gel loader type
Start volume 15 mL 11 mL
Ordering Information
1.5
Concentrator capacity
Dimensions
Materials of construction
Equipment Required
Centrifuge
Concentrate recovery
Recommended tip Thin gel loader type Thin gel loader type
Start volume 15 mL 11 mL
Ordering Information
Vivaspin® Turbo 15 RC 12 pc 48 pc
Technical Specifications
15
10
Concentrator capacity
5
Swing bucket rotor 20 mL
1
0.75
Fixed angle rotor 14 mL
116 0.5
0.2
With pressure head 15 mL
Dimensions
Materials of construction
Equipment Required
Centrifuge
Pressure
Concentrate recovery
Recommended tip Thin gel loader type Thin gel loader type
Start volume 20 mL 14 mL 10 mL 10 mL
Ordering Information
Vivaspin® 20 PES 12 pc 48 pc
Technical Specifications
Concentrator capacity
Swing bucket rotor 90 mL
Device fits standard 250 mL rotors With pressure head 98 mL
Dimensions
Length x diameter 123 x 62 mm
197 x 62 mm with pressure head
Active membrane area 23.5 cm2
Hold-up volume of membrane < 250 μL
Dead-stop volume 350 μL
Materials of construction
Body Polycarbonate (PC)
Filtrate vessel Polycarbonate (PC)
Concentrator cap Polypropylene (PP)
Pressure head Polyoxymethylene (POM) and Aluminium
(ALU)
Pressure head seal Thermoplastic Elastomer (TPE)
Membrane Polyethersulfone (PES)
Equipment Required
Centrifuge
Pressure
-
Low shear, no foaming control Ideal for single samples
Less visual control Use in fridge or cold Faster concentrations
room
Slower concentrations
Ordering Information
0.2
0.5
0.75
1
10
15
0.75
0.75
10
15
0.5
0.2
10
0.5
15
0.2
5
5
1
1
15
10
1
0.75
0.5
0.2
15
10
10
1
0.75
0.5
0.2
15 15
10 10
Pressurize
5 5
1 1
0.75 0.75
0.5 0.5
0.2 0.2
0.1 to 3 L
-
Unique performance
-
The novel Vivaflow® 50 system A single 50 cm2 module will typically
provides a standard of ease of use, reduce 500 mL to less than 15 mL in
performance, flexibility and economy under 50 minutes.
--
which is unrivalled by any laboratory Less than 10 mL minimum system
or pilot scale filtration system on the recirculation for highest
-
market. concentrations.
Less than 500 μL non
Unique features recoverablehold up volume.
Thin channel flip-flow path provides Near total recoveries achievable with
--
high turbulence and cross flow a single 10 mL rinse.
velocities for exceptional flux, even at
high concentrations. Each package of two cassettes
--
No need for pressure holders. contains all of the required tubing and
Crystal clear for simple control and fittings for plug-and-play operation
visibility of membrane status. with a standard peristaltic pump
Unique interlocking modules with accepting 6.4 mm OD (size 16) tubing.
series connectors for easy scale up.
Disposable | single use.
Single cassette
Technical Specifications
Dimensions
Overall L | W | H 25 | 107 | 84 mm
Channel W | H 15 mm | 0.3 mm
Operating conditions
Materials of construction
Ordering Information
--
minutes. Alternatively, speed up your speeding up concentration, two
process by using two Vivaflow® 50R cassettes can be run simultaneously.
cassettes in parallel and concentrate
--
1 liters in under 30 min. Fast and easy protein sample
concentration
Vivaflow® 50R is a plug-and-play Reusable
laboratory crossflow cassette for Concentrates volumes from
-
concentrating up to 1 L aqueous 0.1 L to 1 L
samples. The active membrane area Optimal for concentration of culture
per device is 50 cm2. supernatants and viruses
The most compact crossflow
cassette with a premium Hydrosart®
membrane
Operating conditions
Vivaflow 50R – Two cassettes
®
Pump flow rate 200 – 400 mL/min
Materials of construction
-
Here you can find Start volume Average flux Recovery
instructions on how to 250 mL mL/min
Direct 25 mL rinse
use Vivaflow® 50R for:
-
Lysozyme 0.25 mg/mL (14.3 kDa)
Preparation of biological
5 kDa MWCO HY 70 3.4 96% 98%
nanoparticles and medical 10 kDa MWCO HY 23 10.3 94% 96%
nanocarriers
BSA 1.0 mg/mL (66 kDa)
Concentration and 10 kDa MWCO HY 24 9.9 98% >99%
purification of viruses 30 kDa MWCO HY 15 15.8 97% >99%
Ordering Information
Technical Specifications
Dimensions
Operating conditions
Materials of construction
Ordering Information
0.2 μm VF20P7
Vivaflow 200 HY
®
Technical Specifications
Weight 4.1 kg
Power output 37 W -
Speed | flow rate range 20 – 600 rpm 16 – 480 mL/min (size 16 tubing)
34 – 1,020 mL/min (size 15 tubing)
Materials of construction
Ordering Information
Pump drive (230 V), Easy-Load pump head (size 16), 500 mL 1 VFS502
diafiltration reservoir, cassette stand, spare tubing, T-connectors, series
interconnectors and pressure indicator
Pump drive (115 V), Easy-Load pump head (size 16), 500 mL diafiltration 1 VFS504
reservoir, cassette stand, spare tubing, T-connectors, series
interconnectors and pressure indicator
Pump drive (230 V), Easy-Load pump head (size 16), 500 mL 1 VFS202
diafiltration reservoir, and spare tubing
Pump drive (115 V), Easy-Load pump head (size 16), 500 mL diafiltration 1 VFS204
reservoir, and spare tubing
Technical Specifications
Materials of construction
Ordering Information
Vivaflow® accessories for operating multiple cassettes Pack Size Prod. No.
Vivaflow® tubing set (2x 1 m feed tubing with Luer fitting, 2x 0.5 m 1 VFA034
retentate tubing with flow restrictor, 1x series interconnector)
pH range 1 – 14 4–8 1 – 13
Protein retention
Ordering Information
63 mm 14609--63------D
76 mm 14609--76------D
47 mm 14629--47------D
63 mm 14629--63------D
76 mm 14629--76------D
43 mm 14639--43------D
47 mm 14639--47------D
63 mm 14639--63------D
76 mm 14639--76------D
150 mm 14639-150------D
47 mm 14659--47------D
63 mm 14659--63------D
76 m 14659--76------D
47 mm 14650--47------D
76 mm 14650--76------D
63 mm 14668--63------D
47 mm 14679--47------D
76 mm 14679--76------D
47 mm 14529--47------D
47 mm 14539--47------D
50 mm 14539--50------D
43 mm 14549--43------D
47 mm 14549--47------D
63 mm 14549--63------D
63 mm 14419--63------D
44 mm 14429--44------D
47 mm 14429--47------D
63 mm 14429--63------D
76 mm 14429--76------D
47 mm 14439--47------D
63 mm 14439--63------D
76 mm 14439--76------D
47 mm 14459--47------D
63 mm 14459--63------D
76 mm 14459--76------D
Vivacon® 500 46
Vivacon® 2 49
45
Vivacon® 500
Feature Benefit
Reverse spin
Technical Specifications
Concentrator capacity
Dimensions
Materials of construction
1.0
on individual devices.
Feature Benefit
1.0
Easy to remove re-spin cap Convenient sample handling
mL
2.0
Technical Specifications
Concentrator capacity
Materials of construction
53
Vivaclear Centrifugal Filters
-
Applications
---
able microfiltration devices for the fast Clarification of samples before
and reliable clarification or filtration of loading onto Vivapure® protein
biological samples in the range 100 t o purification spin columns
500 μL. They can be used in fixed Removal of particles and participates
angle rotors accepting 2.2 mL centri- Filtration of plasma and serum
--
fuge tubes. Removal of cells or cell debris
Product Features
--
High-flux polyethersulfone
membrane
0.8 μm pore size
Low hold-up volume (< 5 μL)
Fast and reproducible performance
Filter capacity
Dimensions
Length x diameter 43 x 11 mm
Materials of construction
Equipment Required
Centrifuge
Filtrate recovery
Ordering Information
Spin Spin
Vivapure Maxi H
®
60 – 80 mg 19 mL 10.5 mL
---
Typical Applications
--
Fractionation of protein mixtures prior to 1D or 2D-PAGE
Scouting purification conditions for new protein targets
Removal of endotoxins from monoclonal antibodies
---
Vivapure® Maxi H Preparation of heme moiety from heme containing protein prior to functional
analysis
General protein purification and polishing
-
Detergent removal from protein solutions
Purification of antibodies from serum, ascites or cell culture supernatant
Intermediate sample purification prior to further HPLC | FPLC
Purification of membrane-bound proteins
Vivapure Q Mini H
®
24 48 VS-IX01QH24
Vivapure S Mini H
®
24 48 VS-IX01SH24
Vivapure Maxi
®
Vivapure Q Maxi H
®
8 16 VS-IX20QH08
Vivapure® Maxi H
Vivapure S Maxi H
®
8 16 VS-IX20SH08
Adenovirus Purification 61
Vivapure® Adenopack 20 62
Lentivirus Purification 66
Vivapure® Lentiselect 40 67
59
Vivapure® Virus Purification and Concentration Kits
60 Virus Purification and Concentration Vivapure® Virus Purification and Concentration Kits
Adenovirus Purification
--
Adenopack 20 | 100 | 500 Adenopack Advantages
The Adenopack adenovirus purifi
cation and concentration kits offer Fast and Easy Virus Purification
researchers who need to recover up to Purification completed in 2 hours
3 × 1013 purified recombinant adeno Convenient, over 10 × faster
-
virus particles for invitro transfection a alternative to CsCl density gradient
fast, safe and easy to use solution. The
kits include all reagents and devices Quantitative Yields
necessary for clarification, purification In contrast to CsCl density gradient,
and concentration of adenovirus the complete cell culture is used for
type 5 from HEK293 cell cultures in virus purification and not only the
-
only two hours. These straight forward viral pellet
kits replace time-consuming and
labor-intensive 48 hour CsCl density Flexible Product Range
gradients. Applicable from initial construct
screening to large scale virus
-
Adenopack kits are offered as production
Adenopack 20, Adenopack 100
and Adenopack 500, for the Complete Kit
purification and concentration of Including filtration devices,
adenovirus type 5 from 20 mL to Adenopack units for virus purifica-
-
500 mL cell culture, leading to tion, Vivaspin® and all buffers
1 × 1011- 3 × 1013 purified viral particles.
For each sample volume, the most Low Endotoxin Levels
convenient handling method is High cell viability and infection
offered for ultimate convenience. rates due to endotoxin levels of
< 0.025 EU/mL
To this end, preparations using
Adenopack 20 are pursued in
spin column format in a centrifuge,
Adenopack 100 is a manually
operated kit in syringe filter format*,
and Adenopack 500 is a pump
driven kit.
Virus Purification and Concentration Adenovirus Purification with Vivapure® Adenopack Kits 61
Vivapure® Adenopack 20
The optimal kit for construct screening
Technical Data
Kit specifications
Sample size 20 mL of cell culture
Number of purifications 6 × 20 mL
Virus particles (VP) per mL Typically up to 1 × 1011 – 1012
VP | IU 50–100
Processing time Typically 1 hour
Endotoxin level < 0.025 EU/mL
20 mL syringe 4
10 mL syringe (elution) 2
Elution Buffer 1 × 20 mL
Vivaspin 20 concentrator
®
4
Kit specifications
Sample size 20–200 mL of cell culture
Number of purifications 2 × 20–60 mL
1 × 200 mL
Virus particles (VP) per mL Typically up to 1 × 1013
VP | IU 20–50
Processing time Typically 2 hours
Endotoxin level < 0.025 EU/mL
Technical Data
Kit specifications
Sample size 500 mL of cell culture
Number of purifications 1 × 500 mL
Virus particles (VP) per mL Typically up to 3 × 1013
VP | IU 20–50
Processing time Typically 2 hours
Endotoxin level < 0.025 EU/mL
-
Lentiselect 40 | 500 | 1000 Lentiselect Advantages
The Lentiselect lentivirus purification
and concentration kits offer Fast and Easy Virus Purification
-
researchers who need to recover up to Purification completed in under one
5 × 109 infective lentivirus particles per to six hours, depending on sample
mL for invitro transfection or animal volume
-
studies a fast and easy to use solution. Kit as easy to use as filtration
-
approximately one day for large sample equipment such as ultracentrifuges
volumes, thus reducing the purification
time to only a few hours. High Virus Purity
Achieve pure virus due to a
Lentiselect kits are offered as chromatography purification for your
Lentiselect 40, Lentiselect 500 and experiments instead of a crude and
Lentiselect 1000 for the purification variable cell culture supernatant
and concentration of VSV-G pellet
-
pseudotyped lentivirus from 40 mL to 1
L cell culture, leading to 8 × 108 – 1 × 1010 Optimal for Multiple Virus Construct
purified infective particles. For each Screening
sample volume, the most convenient With Lentiselect 40, four purification
handling method is offered. To this runs can be conducted in parallel
-
end, 40 mL sample volumes are with one kit
processed manually with Lentiselect
40, while Lentiselect 500 and 1000 Complete Kits
are pump driven kits. Including Lentiselect units for
virus purification, Vivaspins for
concentration | buffer exchange and
-
all buffers and syrings necessary
66 Virus Purification and Concentration Lentivirus Purification with Vivapure® Lentiselect Kit
Vivapure® Lentiselect 40
Fast purification of up to 8 × 108 viral particles
Technical Data
Kit specifications
Sample size 40 mL cell culture
Number of purifications 4 × 40 mL
Infective particles (P) per mL Typically up to 3 × 109
VP | IU 5 – 15
Processing time Typically 45 minutes
Endotoxin level < 0.025 EU/mL
10 mL syringe
containing
elution buffer
Lentiselect
Unit
Lentiselect
Unit
Purified virus
Tube containing
24 mL buffer
1. 2. Washing 3. Elute
Technical Data
Kit Specifications
Sample volume 500 mL cell culture
Number of purifications 1 × 500 mL
Infective particles (IP) per mL Typically up to 2–5 × 109*
Processing time Typically up to 3 hours
Endotoxin level < 0.025 EU/mL
Technical Data
Kit specifications
Sample volume 1 L cell culture
Number of purifications 1×1L
Infective particles (IP) per mL Typically up to 4 – 5 × 109*
Processing time Typically up to 6 hours
Endotoxin level < 0.025 EU/mL
71
1. Desalting and Buffer Exchange with Vivaspin®
Centrifugal Concentrators
Introduction The protein sample can then be
Vivaspin® centrifugal concentrators, concentrated again to the desired
with patented vertical membrane level, or the buffer exchange can be
technology, combine fast filtration repeated to reduce the salt
with high recovery of target proteins. concentration even further before a
This makes Vivaspin® the technology final concentration of the protein. This
of choice for desalting or buffer process is called diafiltration. For
exchange, avoiding lengthy dialysis proteins with a tendency to precipitate
steps. at higher concentrations, it is possible
to perform several diafiltration steps in
While proteins are retained by an sequence, with the protein
ultrafiltration membrane, salts can pass concentrated each time to only 5 or
freely through, independent of protein 10×. For example, if a precipitous
concentration or membrane MWCO. protein sample is concentrated to 5×
In consequence, the composition of then diluted back to the original
the buffer in the flow-through and volume, and this process is repeated a
retentate is unchanged after protein further two times, this still results in a
concentration. By diluting the >99% reduction in salt concentration,
concentrate back to the original without over-concentrating the
volume, the salt concentration is protein.
lowered. The concentrate can be
diluted with water or salt-free buffer if
simple desalting is required; however,
it is also possible to dilute the
concentrate with a new buffer, thereby
exchanging the buffering substance
entirely. For example, a 10 mL protein
sample containing 500 mM salt, if
concentrated 100-fold still contains
500 mM salt. If this concentrate is
then diluted 100× with water or salt-
free buffer, the protein concentration
returns to the original level, while the
salt concentration is reduced 100× to
only 5 mM (i.e. a 99% reduction in salt
concentration).
1 2 3 4 5
Vivaspin® 20
Vivaspin® 6
100
80
60
40
20
0
milk powder untreated
Figure 1: Protein recovery (10 µg/mL BSA) with Vivaspin® PES 10 kDa after passivation
80
60
40
20
0
250 ng 500 ng
Figure 2: Protein recovery (250 and 500 ng BSA) with Vivaspin® 2 PES 10 kDa after passivation
pH = 6 pH = 8 pH = 10
66 kDa
45 kDa
Target
protein
31 kDa
22 kDa
f = Flow-through 800 4
Note: Scouting for optimal binding conditions of a SH2 domain expressed in E. coli. 12% reducing
SDS gel, silver stained, shows the sample before purification (s), flow-through (f), wash (w) and
eluate (e) fractions (1 M NaCl) from Vivapure® Q and S Mini spin columns, at the various pH values
tested.
66 kDa
45 kDa
Target
protein
31 kDa
22 kDa
f = Flow-through 800 16
Note: Optimizing elution conditions for a SH2 domain expressed in E. coli, using Vivapure® S Mini
spin column at pH 6. 12% reducing SDS gel, silver stained, shows the sample before purification (s),
flow through (f), wash (w) and eluate (e1-5) fractions.
-
Conclusion The results obtained in this experiment
A two-step procedure was used to can be used to various ends, e.g:
--
rapidly scout optimal purification Polishing a specific protein after
conditions for a target protein (a SH2 purification with another chromato-
domain from E. coli lysate) with ion graphic technique
exchange chromatography. In the first Quickly establishing a FPLC method
step, the most suitable ion exchanger for a new protein
and buffer pH for binding the target Identification of the optimal purifica-
protein was verified. In the second step, tion method prior to scale up with
the elution condition was optimized, Vivapure® IEX Maxi spin columns.
building on the results gained in step
one. With the scouting procedure For these purposes Vivapure® IEX Mini
described here, it was possible to and Maxi spin columns and Sartobind®
quickly and conveniently purify the membrane adsorber units with FPLC
target protein to homogeneity. connectors are available.
Gene therapy for Concentration and Vivaspin® 20 Titer, ELISA, cell binding 8
cancer treatment purification after (1,000 kDa) assay, apoptosis | cell cycle
(adeno-associated expression, Buffer assay
virus; rAAV-2, human) exchange after
His tag (FreeStyle 293
Expression Medium
(Gibco), serum-free)
Study on host genome Clarification with 0.2 μm Vivaflow® 200 CsCl gradients, 29
integration (virophage filter and concentration (0.2 μm and electron microscopy
mavirus, Cafeteria with 100 kDa filter 100 kDa)
roenbergensis) (Cafeteria roenbergen-
sis, f/2 medium)
Investigation into the role Concentration of VLPSs Vivaspin®, PES Cell-cell fusion 36
of three transmembrane from HEK 293T cell assay
proteases in the activa- culture supernatant
tion of SARS-CoV-1 spike
(S) protein
Investigation of viral and Concentration of SARS- Vivaspin®, PES Western blot 41,
cellular determinants CoV and hCoV-EMC analysis 42
governing hCoV-EMC virus like particles (VLPs)
entry into host cells
Measurement of SARS- Concentration of viral Vivaspin®, PES Viral RNA extraction 43,
CoV-2 RNA in sewage RNA (50 kDa) and purification RT- 44,
qPCR quantification 45
Virus detection in full scale Concentration of viral Vivaflow® 50, PEG precipitation 48
membrane bioreactor particles in effluent PES Viral
(MBR) plant by virus RNA quantification
concentration monitoring,
inc. Norovirus, Sapovirus
and Rotavirus
Table 1 summarizes examples of Nanocarrier ultrafiltration applications with Sartorius Vivaspin® or Vivaflow®:
Iron oxides nanoparticles with SD: 4.5 ± 0.9 nm via X-Ray-Diffraction Analysis Magnetic resonance imaging 7
cisplatinbearing polymer coating
Functionalized iron oxide nanoparticles SD: 38 and 40 nm via DLS Stem cell gene therapy and tracking 8
Gold nanoparticles SD: 0.8 – 10.4 nm via Atomic Force Microscopy Antimicrobial activity 17
Protein coated gold nanoparticles SD: 15 and 80 nm via TEM Drug delivery 18
Functionalized gold nanoparticles Core-SD: 2 nm via TEM Targeted imaging tool and antigen delivery 19
Functionalized gadolinium-based Z-Average: 1.1 ± 0.6 nm and 4 – 14 nm Diagnostic and therapeutic application 20, 21
nanoparticles
Curdlan coated polymer nanoparticles Z-Average: 280 – 480 nm depending on the Macrophage stimulant activity 23
composition and drug delivery
Liposomes and micelles Z-Average: 100 nm for Liposomes and 15 nm Ischemia-reperfusion injury 25
for micelles
Solid lipid Nanoparticles Z-Average: 100 – 120 nm depending on the Drug delivery (Brain Targeting) 26
used lipid
Bacterial outer membrane vesicles SD: 124 nm via TRPS Tunable resistive pulse sensing (TRPS) 27
Analysis
Bacterial outer membrane vesicles SD: 50 – 150 nm via TEM Basic research 30
Micelles
Protein Nanoparticles
SD = Size distribution
Iron oxides nanoparticles with Vivaspin® 20 100 kDa Purification and concentration 7
cisplatinbearing polymer coating
Protein coated gold nanoparticles Vivaspin® 6 10 kDa Separation of Nanoparticles | Dyes and washing 18
Docetaxel-carboxymethylcellulose Vivaspin ®
10 kDa Concentration 4
Polymer Nanoparticles
Bacterial outer membrane vesicles Vivaflow® 200 100 kDa Buffer exchange and concentration 27
Bacterial outer membrane vesicles Vivaspin® 500 and 20 100 kDa Buffer exchange and concentration 28
Liposomes Vivaspin ®
100 kDa External buffer exchange 2
Micelles
Protein Nanoparticles
Protein Nanoparticles Vivaspin® 500 3 kDa Separation of the free from the encapsulated drug 32
(Drug binding quantification by subsequent
UV-vis analysis)
1,200 0:50:36
1,300 0:55:32
1,400 1:00:24
1,500 1:05:26
1,600 1:10:28
1,700 1:15:52
1,800 1:21:50
Figure 3: Electron micrograph of chromatogra- Figure 4: Vivapure® Maxi spin columns can be
phy gel beads (upper right) in comparison to a used in a centrifuge for fast and easy protein
Q ion exchange membrane adsorber (back- purification.
ground), revealing 100-fold larger pore sizes of
the membrane adsorber.
F
or further information, visit
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