Bio-Scale™ Mini UNOsphere SUPrA™ Cartridges, 1 and 5 ml
Instruction Manual
Catalog # 732-4200 732-4201 732-4202
For technical support, contact your local Bio-Rad office, or in the U.S., call 1-800-4BIORAD (1-800-424-6723).
Table of Contents
Section 1 |
...Introduction. ....................................... |
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Section 2...Connection to Bio-Rad’s Low- |
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Pressure Chromatography |
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Instruments ........................................ |
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Section 3 .. |
Connection to Other Liquid |
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Chromatography Systems ................ |
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3.1 |
BioLogic DuoFlow™ |
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Systems................................. |
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3.2 |
HPLC Systems ...................... |
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3.3 |
FPLC Systems ....................... |
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Section 4 .. |
Getting Started................................. |
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4.1 |
Screening Buffers and |
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Conditions for UNOsphere |
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SUPrA.................................... |
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4.2 |
Scaling Up the Separation ..... |
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Section 5 .. |
Care of the Cartridge........................ |
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5.1 |
Cleaning................................. |
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5.2 |
Sanitation................................... |
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5.3 |
Autoclaving ............................ |
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5.4 |
Storage.................................. |
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Section 6 .. |
Technical Assistance ........................ |
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Section 7 .. |
Ordering Information......................... |
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Section 8 .. |
References ....................................... |
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Section 1
Introduction
Bio-Scale™ Mini cartridges have a patent-pending, double-wall design that provides extra durability and allows easy, reliable runs with aqueous buffers most commonly used for protein purification. The polypropylene Luer fittings and internal sealing surfaces assure leak-free operation at pressures up to 45 psi. The cartridges are convenient, disposable, and supplied ready for use. Cartridges are available for a variety of chromatographic techniques including desalting, ion exchange, and affinity chromatography. See Ordering Information (Section 7) for a listing of the complete Bio-Scale Mini cartridge product line.
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UNOsphere SUPrA™ medium is a chromatographic support based on recombinant protein A. The media are designed for process-scale purification of monoclonal antibodies. The protein A ligand is produced in E. coli without the use of material from animal origin. The UNOsphere base bead is a macroporous polymeric bead that is designed for robust and scaleable applications. See Tables 1 and 2 on pages 4–5 for the technical description of the product.
UNOsphere SUPrA media are built on the proven UNOsphere base bead, which insures an easy scale-up path for process applications. The outstanding flow pressure performance of UNOsphere chromatography media allow its use in process applications without concern for exceeding the pressure limits of the media or chromatography system. The flow characteristics of UNOsphere SUPrA packed in a large column format are shown in Figure 1.
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UNOsphere SUPrA affinity chromatography media come with full regulatory support and are backed by the support of Bio-Rad’s global application and development team.
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3.0 |
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2.5 |
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2.0 |
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Pressure |
1.5 |
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1.0 |
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0.5 |
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0.0 |
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Linear Flow (cm/hr)
Fig. 1. Flow performance of UNOsphere SUPrA media in Bio-Rad's EasyPack™ column (20 x 20 cm) packed to 13.1% axial compression.
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Table 1. Bio-Scale Mini UNOsphere SUPrA
Cartridge Specifications.
Sizes |
1 and 5 ml bed volumes |
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Dimensions |
1 ml: |
40 x 5.6 mm, length x inner |
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diameter |
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5 ml: |
40 x 12.6 mm, lenghth x inner |
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inner diameter |
Maximum pressure tolerance |
45 psi |
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Recommended flow rates |
1 ml: 1–2 ml/min (240–480 cm/hr) |
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5 ml: 5–10 ml/min (140–480 cm/hr) |
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Maximum flow rate |
1 ml: 3 ml/min (730 cm/hr) |
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5 ml: 15 ml/min (722 cm/hr) |
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Fittings |
Female Luer fitting inlet and |
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male Luer fitting outlet |
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Column material |
Polypropylene |
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Frit material |
Polyethylene (HDPE) |
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Shipping condition |
20% ethanol |
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Storage recommendation |
20% ethanol |
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Autoclavability |
Not autoclavable |
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Table 2. Technical Description of UNOsphere
SUPrA Affinity Chromatography Media.
Composition |
Highly crosslinked polyacrylamide polymer |
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Particle size range |
53–61 µm |
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Ligand |
Recombinant protein A |
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Coupling chemistry |
Epoxy |
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Dynamic binding capacity* |
150 cm/hr |
30 ± 3 mg/ml |
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300 cm/hr |
25 ± 2 mg/ml |
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450 cm/hr |
20 ± 2 mg/ml |
Chemical stability** |
10 mM hydrochloric acid |
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6 M guanidine hydrochloride |
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0.1 M arginine (pH 2.8) |
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0.1 M citrate (pH 2.8) |
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0.1 M glycine |
(pH 2.8) |
Working pH range |
3–11 |
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Cleaning-in-place (CIP) |
6 M guanidine hydrochloride |
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10 mM hydrochloric acid |
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0.1 M sodium hydroxide |
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1 M acetic acid/20% ethanol |
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Recommended mobile phase |
100–600 cm/hr |
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velocity range |
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Temperature stability |
4–40°C |
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Delivery conditions |
50% slurry in 20% ethanol |
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Storage conditions |
4–8°C |
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*Minimum 20 mg/ml at 300 cm/hr; 10% breakthrough capacity determined with 1.0 mg/ml polyclonal human IgG in 1.1 x 10 cm column.
**No significant change in chromatographic performance after 24-hour storage at room temperature.
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