Bio-Rad Bio-Scale Mini UNOsphere SUPrA Affinity Cartridges User Manual

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. .......................................1

Section 2...Connection to Bio-Rad’s Low-

..............Pressure Chromatography

..............Instruments ........................................5

Section 3 ..Connection to Other Liquid

..............Chromatography Systems................10

..............3.1 BioLogic DuoFlow™

Systems.................................11

3.2 HPLC Systems ......................11

3.3 FPLC Systems .......................12

Section 4 ..Getting Started.................................13

4.1 Screening Buffers and
Conditions for UNOsphere

SUPrA....................................14

4.2 Scaling Up the Separation .....16

Section 5 ..Care of the Cartridge........................19

5.1 Cleaning.................................19

5.2 Sanitation...................................

5.3 Autoclaving ............................21

5.4 Storage..................................21

Section 6 ..Technical Assistance ........................22

Section 7 ..Ordering Information.........................23

Section 8 ..References .......................................25

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.

1

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.

2

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.

Fig. 1. Flow performance of UNOsphere SUPrA media in
Bio-Rad's EasyPack™ column (20 x 20 cm) packed to 13.1%
axial compression.

0.0

0.5

1.0

1.5

2.0

2.5

3.0

0 100 200 300 400 500 600 700

3

Pressure (Bar)

Linear Flow (cm/hr)

Table 1. Bio-Scale Mini UNOsphere SUPrA
Cartridge Specifications.

Sizes 1 and 5 ml bed volumes

Dimensions 1 ml: 40 x 5.6 mm, length x inner

diameter

5 ml: 40 x 12.6 mm, lenghth x inner

inner diameter

Maximum pressure tolerance 45 psi

Recommended flow rates 1 ml: 1–2 ml/min (240–480 cm/hr)

5 ml: 5–10 ml/min (140–480 cm/hr)

Maximum flow rate 1 ml: 3 ml/min (730 cm/hr)

5 ml: 15 ml/min (722 cm/hr)

Fittings Female Luer fitting inlet and

male Luer fitting outlet

Column material Polypropylene

Frit material Polyethylene (HDPE)

Shipping condition 20% ethanol

Storage recommendation 20% ethanol

Autoclavability Not autoclavable

4

Table 2. Technical Description of UNOsphere
SUPrA Affinity Chromatography Media.

Composition Highly crosslinked polyacrylamide polymer

Particle size range 53–61 µm

Ligand Recombinant protein A

Coupling chemistry Epoxy

Dynamic binding capacity* 150 cm/hr 30 ± 3 mg/ml

300 cm/hr 25 ± 2 mg/ml
450 cm/hr 20 ± 2 mg/ml

Chemical stability** 10 mM hydrochloric acid

6 M guanidine hydrochloride

0.1 M arginine (pH 2.8)

0.1 M citrate (pH 2.8)

0.1 M glycine (pH 2.8)

Working pH range 3–11

Cleaning-in-place (CIP) 6 M guanidine hydrochloride

10 mM hydrochloric acid

0.1 M sodium hydroxide
1 M acetic acid/20% ethanol

Recommended mobile phase 100–600 cm/hr
velocity range

Temperature stability 4–40°C

Delivery conditions 50% slurry in 20% ethanol

Storage conditions 4–8°C

* 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.

5