GE Q HP, SP HP 3. Operation

p. 9

Fig 3. Recommended buffer substances for anion exchange chromatography.

The columns can be operat ed with a syringe, p eristaltic pum p or a

chromatography system.

3. Operation

Buﬀ er preparation

Water and chemicals used f or buffer prepar ation should be of hi gh purity. It

is recommended to fil ter the buffers b y passing them thr ough a

0.22 μm filter immedia tely before use. See Tabl es 2 and 3, Figs 2 and 3 for

recommended buffers.

Sample preparation

The sample should be ad justed to the compo sition of the star t buffer.

This can be done by eithe r diluting the sam ple with start b uffer or by

buffer exchange using HiTrap Desalting, HiPrep™ 26/10 Desalting or PD-10

column. The sample sho uld be filtered t hrough a 0.22 μm or 0.45 μm fil ter or

centrifuged immed iately before it is ap plied to the column . See Table 4.

Contents

Main Instructions 71-7149-00 AN HiTrap ion exchange columns GE Healthcare HiTrap SP HP, HiTrap Q HP, Connectorkit Table of contents 1. Description Media properties Column p. 4 Table 1. HiTrap SP HP and HiTrap Q HP column s characteris tics 2. Selection of ion exchanger and conditions Selection of bu er pH and ionic strength Strategy 1. Binding and elution of all sample components Strategy 2. Enrichment of target protein Strategy 3. Binding of sample contaminants Start bu er Starting pH p. 8 Table 3. Buffers for anion exchange chromatography. Fig 2. Recommended buffer s ubstances for ca tion exchange c hromatograph y. pH 3. Operation Bu er preparation Sample preparation p. 10 Table 4. Prepacked column s for desalting an d buffer exchang e. Puri cation 4. Optimization Further optimization 5. Choice of gradient type Elution with stepwise ionic strength gradients Elution with continuous ionic strength gradients 6. Determination of binding capacity Page 7. Scaling up 8. Storage 9. Ordering Information Page Page Elanders stervla 2006