Bio-Rad Bio-Scale™ CHT™ Type I Columns - Manual (Page 4)

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Table 1. Column Characteristics CHT2-I
CHT2-I CHT5-I CHT10-I CHT20-I
Column volume (ml) 2 5 10 20
Recommended max.
protein loading (mg)
20 50 100 200
Recommended flow
arates (ml/min)
0.5 to 3.0 0.5 to 5.0 0.5 to 7.0 0.5 to 10.0
Dynamic protein binding
capacity (mg Lysozyme)/
column
30 75 150 300
Static DNA binding
capacity (mg calf thymus
DNA)
0.20 0.5 1.0 2.0
Average particle size (µm) 10 ± 3.0 10 ± 3.0 10 ± 3.0 10 ± 3.0
Column dimensions (mm) 7 x 52 10 x 64 12 x 88 15 x 113
Maximum operating
pressure (psi/bar)
1,000/67 750/50 600/40 500/34
Note: Bio-Rad Laboratories does not recommend nor warranty the use of the Bio-Scale
columns with solvent delivery systems containing stainless steel parts and used with corrosive
eluents containing e.g. halide salts. We recommend the use of inert, biocompatible (ceramic,
PEEK, titanium) solvent delivery systems for maximum column life and recovery of sample
biological activity.
Section 2
Use of the Bio-Scale CHT-I Columns
2.1 Preparation for Initial Use
The columns are supplied in a storage solution of 20% ethanol in 5 mM phosphate buffer,
pH 6.8. Prior to initial use and after extended storage periods, each column should be condi-
tioned as described below (steps 1-4). Always use HPLC grade reagents, and filter and degas
buffers). During this operation do not exceed more than 25% of the recommended maximum
flow rates (see Table 1).
1. Wash with 5 column volumes of water.
2. Wash with 5 column volumes of low ionic strength equilibration buffer. Typically 5-10
mM sodium phosphate pH 6.8
3. Wash with 5 column volumes of high ionic strength limit buffer. Typically 500 mM sodium
phosphate pH 6.8.
4. Wash with 5 column volumes of low ionic strength equilibration buffer.
The column may now be further equilibrated in the starting buffer at the desired flow
rate.
Always use buffer components of the highest available purity as UV absorbing impurities
may cause baseline disturbances that interfere with the detection of protein peaks.
2.2 Sample Preparation
Proper adjustment of the sample pH and ionic strength is critical for consistent and repro-
ducible chromatography. For best results, the sample should be exchanged into the start buffer