Bio-Rad UNOsphere Rapid S Media User Manual

Bio-Scale™ Mini UNOsphere™ Q, S and Rapid S Cartridges, 1 and 5 ml

Instruction Manual

Catalog #

732-4100, 732-4102, 732-4400,

732-4104, 732-4110, 732-4401,

732-4112, 732-4114, 732-4402

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 ..Preparing a Cartridge For Use..........13

..................4.1 Sample Preparation ...............14

..................4.2 General Purification Protocol..15

..................4.3 Scaling Up the Separation .....16

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

..................5.1 Autoclaving ............................20

..................5.2 Storage..................................20

Section 6 ..Technical Assistance ........................21

Section 7 ..Ordering Information.........................22

Section 8 ..References .......................................24

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. The design of the Bio-Scale Mini cartridges offers:

• Ready-to-go convenience; simply equilibrate the cartridge in the buffer of choice

• Luer fittings for convenient connection to any chromatography system or directly to a Luer-Lok syringe

The Bio-Scale Mini UNOsphere™ Q, S, and Rapid S cartridges are packed with UNOsphere ion exchange supports. These supports are based on hydrophilic spherical, polymeric beads designed for the purification of proteins, nucleic acids, viruses, plasmids, and other macromolecules. UNOsphere beads are designed to provide high capacity, low back-pressure, and high productivity. Detailed product information is given in Tables 1 and 2.

Table 1 Bio-Scale Mini UNOsphere Q, S, & Rapid S Cartridge Specifications.

Sizes 1 and 5 ml bed volumes

Dimensions 1 ml: 40 x 5.6 mm inner diameter

5 ml: 40 x 12.6 mm 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: 4 ml/min (970 cm/hr)

5 ml: 20 ml/min (963 cm/hr)

Fittings Female Luer fitting inlet and

Column material Polypropylene

Frit material Polyethylene (HDPE)

Shipping condition 20% ethanol

Storage recommendation 20% ethanol

Autoclavability Not autoclavable

male Luer fitting outlet

Table 2 UNOsphere Q, S, and rapid S Specifications

UNOsphere Q UNOsphere S UNOsphere Rapid S

Type of ion exchanger Strong anion Strong cation Strong cation

Functional group -N+(CH

Total ionic capacity 120 µeq/ml Ni2+260 µeq/ml 140 µeq/ml

Dynamic binding capacity*

At 150 cm/hr 180 mg BSA/ml 60 mg IgG/ml 60 mg/ml At 600 cm/hr 125 mg BSA/ml 30 mg IgG/ml 30 mg/ml

Shipping counterion Cl

Median particle size 120 µm 80 µm 100 µm

Recommended 50–1,200 cm/hr 50–1,200 cm/hr 50–800 cm/hr linear flow rate

Chemical stability

1.0 M NaOH (20ºC) Up to 10,000 hr Up to 10,000 hr Up to 800 hr

1.0 M HCl (20ºC) Up to 200 hr Up to 200 hr Up to 200 hr

pH stability 1–14 1–14 1–14

Antimicrobial agent 20% ethanol 20% ethanol 20% ethanol

Regeneration 1–2 M NaCl 1–2 M NaCl 1–2 M NaCl

Storage conditions 20% ethanol or 20% ethanol or 20% ethanol or

* 10% breakthrough capacity determined with 4.5 mg/ml human IgG (UNOsphere

Rapid S) and 2.0 mg/ml BSA (UNOsphere Q) in a 1.1 x 20 cm column.

3)3

0.1 M NaOH 0.1 M NaOH 0.1 M NaOH

-SO

Section 2 Connecting to Bio-Rad's Low-Pressure Chromatography Instruments

The Bio-Scale™ Mini cartridges are ideal for use with Bio-Rad's BioLogic™ LP system, Econo™ Gradient Pump, and Model EP-1 Econo pump, and all lowpressure chromatography instruments. Bio-Scale Mini cartridges can be conveniently connected directly to the system using the Luer lock fittings on the cartridge.

1. Install 1.6 mm ID tubing in the pumphead.

Adjust platen pressure screw (on pumphead). Using a screwdriver or coin, turn the screw counterclockwise as far as it will go, then turn clockwise three full turns. Assemble with fittings and lock rings as shown in Figure 1.

(Use orange lock rings and medium size barb fittings with 1.6 mm tubing.)

Platen pressure

screw

See detail

2 3

456

7 8

C .

Alarm

Fig. 1. Biologic LP setup

Lock ring

Tubing

Luer fitting

+ 22 hidden pages