Bio-Rad ENrich High-Resolution Ion Exchange Columns User Manual

ENrich™ Q ENrich

™

S High-Resolution Ion Exchange Columns

Instruction Manual

Catalog numbers

780-0001 780-0003 780-0021 780-0023

Please read these instructions before you use ENrich Q and S high-resolution ion exchange media. If you have any questions or comments regarding these instructions, please contact your Bio-Rad Laboratories representative.

Table of Contents

Section 1: Characteristics of the ENrich Ion

Exchange (IEX) Columns ................................... 1

1.1 Introduction .......................................................1

1.2 The ENrich Separation Media ............................. 1

1.3 Connection to the NGC™ and Other

Chromatography Systems ................................. 2

Section 2 : Use of the ENrich IEX Columns ........................ 3

2.1 Preparation for Initial Use ................................... 3

2.2 Buffer Selection ................................................. 3

2.3 Sample Preparation ........................................... 5

2.4 Elution Conditions .............................................. 5

Section 3 : Care of the ENrich IEX Columns ....................... 7

3.1 Column Cleaning ............................................... 7

3.2 Bed Height Adjustment ...................................... 8

3.3 Frit Replacement ............................................... 9

3.4 Storage Conditions ...........................................12

Section 4 : Product Information .........................................13

Section 1: Characteristics of the ENrich Ion Exchange Columns

1.1 Introduction

ENrich prepacked ion exchange columns meet the needs of the biochromatographer for rapid and reproducible high-resolution separations of proteins and other biomolecules, including peptides and polynucleotides. Two column sizes (bed volumes of 1 and 8 ml) provide predictable scale-up of high-resolution separations without sacriﬁcing capacity.

1.2 The ENrich Separation Media

Each column contains a spherical, rigid, and highly porous polymeric support derivatized with the strongly basic –N+(CH3)3 quaternary ammonium group or the strongly acidic –SO The 10 μm particle-size and narrow particle size distribution provide excellent resolution of biomolecules at high ﬂow rates and with low backpressures. The hydrophilic ENrich media demonstrate extremely low non-speciﬁc binding of biomolecules accompanied by high recovery of biological activity.

Stability

The columns are stable over a 2–12 pH range, allowing easy cleaning and regeneration.

The ENrich support is compatible with aqueous solutions of 6 M guanidine-HCl and 8 M urea. Nonionic detergents and organic solvents such as methanol, ethanol, and isopropanol may also be used. Ionic detergents can be used only if the detergents have the same charge as the support (that is, sodium dodecyl sulfate (SDS), a negatively charged detergent, should not be used with a positively charged anionic exchanger like ENrich Q).

–

group.

ENrich Instruction Manual 1

Table 1. Column characteristics.

Q S

Column dimensions (diameter x height, mm)

Column volume, ml 1 8 1 8

Maximum protein capacity, mg

Bovine serum albumin 130 1000 – –

Human IgG – – 120 940

Average particle size, μm 10 10 10 10

Maximum operating pressure, psi

Recommended ﬂow rate, ml/min*

* At room temperature. Viscosity may increase at lower temperatures, which will reduce the recommended ﬂow rates.

5 x 50 10 x 100 5 x 50 10 x 100

500 500 500 500

1 4 1 4

1.3 Connection to the NGC and Other Chromatography Systems

The ENrich columns are ﬁtted with 10-32 type female ﬁttings on either end. Standard 10-32 ﬁttings can be used to plumb the column to Bio-Rad’s NGC chromatography system or other vendors’ systems. Adaptors are available for connection to systems that use 1/4-28 ﬁttings (catalog # 750-0564).

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