Filtration of Reagents (> 50 mL) with a VacuCap® Vacuum Filtration Device 2.5.5
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Particulate Removal Sections
Filtration of Reagents (> 50 mL) with a VacuCap® Vacuum Filtration DeviceUncharged microporous membrane filters remove particles from a fluid stream by the sieving or screening mechanism. Particles larger than the pore diameter do not pass through the filter. Such membranes initially show at least 2 to 3 log removal of particles which are equal to or larger than the rated pore size. There are two classic types of microfiltration (MF) processes:
When fluid passes through the filter, particles larger than the spaces within the filter matrix are retained, accumulating primarily on the surface of the filter. The distinction between filters is important because the two classes serve very different functions. Depth filters are usually used as prefilters because they are an economical way to remove 98% of suspended solids and protect elements downstream from fouling or clogging. Screen or microporous filters remove 99.99% of suspended solids and may be used as either prefilters or clarifying filters. |
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Larger volume (> 50 mL) reagents used in analysis can be clarified by microfiltration in the VacuCap and VacuCap PF vacuum filtration devices. These devices employ Supor® hydrophilic polyethersulfone 0.1, 0.2, and 0.45 μm pore size membranes which offer high flow rates and throughputs, and are ideal for solutions where low protein binding is required. In addition, PF devices with 0.8/0.2 μm pore size combination membranes offer high filtration throughput in sample streams with high numbers of particles. The VacuCap and VacuCap PF device family properties are summarized in Table 2.60 for 60 and 90 mm diameter devices.
Figure 2.58
VacuCap Filtration Devices
Table 2.60
Properties of the VacuCap® and VacuCap PF Vacuum Filtration Devices
Vacuum filtration devices have a wide variety of applications and are key to some analytical and screening processes in biosciences. Typical applications of VacuCap MF clarification in proteomics includes:
Media preparation for cell culture applications can be a tedious process, depending on the volume and viscosity of the material to be filtered. The sterile VacuCap vacuum filtration device was engineered for this purpose. PF devices, designed with a built-in prefilter, use the ingenuity of the standard VacuCap device and increase its throughput performance with serum-containing media or other viscous solutions. Other applications requiring 0.2 μm filtration will also see the throughput benefits of VacuCap PF devices when encountering high particle load in a solution. A summary of vacuum filtration of tissue culture media and calf serum is presented in Table 2.61 for the VacuCap 90 mm device with the 0.2 μm and 0.8/0.2 μm PES membranes. The data clearly shows rapid filtration of high viscosity fluids, such as tissue culture media with the PES membranes, in this bottle-top filtration format. This is a useful configuration for a wide range of large volume liquid samples used in analysis.Table 2.61
VacuCap 90 PF Device Filtration Efficiency1
VacuCap 60 Devices, Supor® Membrane
VacuCap 90 Devices, Supor® Membrane
Figure 2.58
VacuCap Filtration Devices Table 2.60
Properties of the VacuCap® and VacuCap PF Vacuum Filtration Devices | Specification | Parameter |
| Membranes Supor® PES |
Hydrophilic polyethersulfone |
| Device Membrane Housing Inlet Tubing Sinker on Inlet Tubing Sealing Gasket |
Polyester Modified Acrylic Medical grade PVC Glass-filled polyurethane elastomer Silicone rubber |
| Effective Filtration Area 60 mm 90 mm. |
30 cm2 60 cm2 |
| Pore Size | 0.1, 0.2, 0.45, 0.8/0.2 μm |
| Sample Volume 60 mm 90 mm |
0.5-1.0 L 1.0-5.0 L |
| Hold-up Volumes | 3.4 mL |
| Maximum Temperature | 55ºC |
| Maximum Vacuum | 63.5 cm Hg (25 in. Hg) |
| Typical Water Flow Rate (mL/min) at 25.4 cm Hg (10 in. Hg) 60 mm (0.1 μm) 60 mm (0.2 μm) 60 mm (0.45 μm) 60 mm (0.8/0.2 μm) 90 mm (0.1 μm) 90 mm (0.2 μm) 90 mm (0.45 μm) 90 mm (0.8/0.2 μm) |
50 200 280 200 100 400 560 400 |
| Collection Vessel 60 mm 90 mm |
Can be used on receptacles with openings ranging from 60 mm, 2-5 cm (0.8-1.9 in.) and 90 mm, 2-6.5 cm (0.8-2.5 in.) |
Application Guidelines for Filtration of Reagents (> 50 mL) with a VacuCap® Vacuum Filtration Device
Vacuum filtration devices have a wide variety of applications and are key to some analytical and screening processes in biosciences. Typical applications of VacuCap MF clarification in proteomics includes:
- Filter sterilization of components or additives for cell culture that are heat labile.
- Clarification of reagents and stains used in proteomics.
Tip: All bulk solid reagents contain a variable percent of their content as in-soluble solids. - Stains and detection reagents used in electrophoresis and Western blotting to remove insoluble solids.
Tip: Many of these reagents are colored or somewhat opaque and it is not possible to see in-soluble material in suspension or as precipitates in their containers. This can lead to nonspecific background staining or detection.
Protocol for Filtration of Reagents (> 50 mL) with a VacuCap Vacuum Filtration Device
A. Materials Required
- VacuCap filtration devices with MF membranes. For specifications, see Table 2.60.
- Vacuum source [max 63.5 cm Hg (25 in. Hg)].
B. VacuCap MF Filtration of > 50 mL Samples
- Remove VacuCap device from the sterile packing and place with gasket seated on the rim of the receiving bottle.
- Connect the feed tubing to the port marked "INLET" on the top of the device. Place the opposite end of the feed tubing in the vessel from which the unfiltered sample will be drawn.
Tip: Very viscous solutions, such as serum-containing culture media, may prematurely clog the VacuCap device. If the device does clog before the entire batch can be filtered, try the VacuCap PF (with 0.8 μm PES prefilter) device. - Connect the vacuum tubing to the port marked "VACUUM" on the top of the device. Refer to product insert for safety precautions.
- While holding the device securely on the receiving bottle, start the vacuum. The device will seal securely to the receiving bottle and fluid will be drawn.
- When filtration is complete, switch off the vacuum pump and allow the vacuum inside the receiving bottle to dissipate.
Tip: These filtration devices are for SINGLE USE ONLY.
Application Data for Filtration of Reagents (> 50 mL) with a VacuCap® Vacuum Filtration Device
Media preparation for cell culture applications can be a tedious process, depending on the volume and viscosity of the material to be filtered. The sterile VacuCap vacuum filtration device was engineered for this purpose. PF devices, designed with a built-in prefilter, use the ingenuity of the standard VacuCap device and increase its throughput performance with serum-containing media or other viscous solutions. Other applications requiring 0.2 μm filtration will also see the throughput benefits of VacuCap PF devices when encountering high particle load in a solution. A summary of vacuum filtration of tissue culture media and calf serum is presented in Table 2.61 for the VacuCap 90 mm device with the 0.2 μm and 0.8/0.2 μm PES membranes. The data clearly shows rapid filtration of high viscosity fluids, such as tissue culture media with the PES membranes, in this bottle-top filtration format. This is a useful configuration for a wide range of large volume liquid samples used in analysis.
Table 2.61
VacuCap 90 PF Device Filtration Efficiency1| Filtrate Volume (mL) | ||||
| Sample | 1.5 min. | 3.0 min. | 5.0 min. | 6.0 min. |
| Tissue Culture Fluid (RPMI + 10% calf serum) 0.2 μm Supor® PES 0.8/0.2 μm Supor PES |
950 950 |
2200 2200 |
ND ND |
3600 3600 |
| Whole Calf Serum 0.2 μm Supor PES 0.8/0.2 μm Supor PES |
ND ND |
ND ND |
160 320 |
ND ND |
Filtration conducted at 21 °C (70 °F) with 50 cm Hg (20 in. Hg) vacuum for a 5 minute filtration time. Actual results may vary depending upon type and concentration of serum, liquid temperature, and applied vacuum.
Ordering Information for Filtration of Reagents (> 50 mL) with a VacuCap® Vacuum Filtration Device
VacuCap 60 Devices, Supor® Membrane
| Part Number | Description | Pkg |
| 4631 | 0.1 μm, 60 mm, sterile | 10/pkg |
| 4632 | 0.2 μm, 60 mm, sterile | 10/pkg |
| 4634 | 0.45 μm, 60 mm, sterile | 10/pkg |
| TA4632* | 0.2 μm, 60 mm, sterile | 10/pkg |
| 4638 | PF 0.8/0.2 μm, 60 mm, sterile | 10/pkg |
| Part Number | Description | Pkg |
| 4621 | 0.1 μm, 90 mm, sterile | 10/pkg |
| 4622 | 0.2 μm, 90 mm, sterile | 10/pkg |
| 4624 | 0.45 μm, 90 mm, sterile | 10/pkg |
| TA4622* | 0.2 μm, 90 mm, sterile | 10/pkg |
| 4628 | PF 0.8/0.2 μm, 90 mm, sterile | 10/pkg |