Macro-Prep DEAE Media

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Overview

Macro-Prep DEAE Dynamic Binding Capacity

The effect of flow rate on the dynamic binding capacity of the Macro-Prep DEAE support. A sample containing 3 mg/ml BSA was separated on a 1 x 10 cm (3 ml) Macro-Prep DEAE column. Sample was loaded onto the column in 10 mM Tris-HCl, pH 7.6 (buffer A) and was eluted in 1 M NaCl in buffer A. Sample was applied until the eluting buffer had an A₂₈₀ value 50% of the incoming sample.

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Scalability of Purification

Macro-Prep DEAE Support: Scaling Up Purification Process

Scalability of separations on Macro-Prep DEAE support. Separation of a yeast enzyme concentrate (Sigma) under various conditions of scale. A, a 50 mg sample was separated on a 7.4 ml column (10 mm ID) using a flow rate of 78 cm/hr (1 ml/min) in an analytical flow cell; B, a 500 mg sample was separated on a 64 ml column (25 mm ID) using a flow rate of 78 cm/hr (6.4 ml/min) in an analytical flow cell; C, a 1 g sample was separated on a 64 ml column (25 mm ID) using a flow rate of 305 cm/hr (25 ml/min) in a preparative flow cell. For all separations, sample was loaded in 50 mM Tris-HCl, pH 8.3, washed with 1 column volume of the same buffer, and eluted in a linear gradient of 0–0.35 M NaCl in the same buffer for 22.5 column volumes.

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DEAE Flow Rate Properties

Macro-Prep^® DEAE Support: Effects of Flow Rate

The effect of flow rate on separation and peak symmetry. A 5 ml sample containing 3.65 mg/ml each myoglobin, conalbumin, BSA, and soybean trypsin inhibitor was loaded onto a column containing 6.56 ml Macro-Prep DEAE support. The sample was loaded in 50 mM Tris-HCl, pH 7.6 (buffer A) and eluted in a gradient of 0–1 M NaCl in buffer A as follows: buffer A for 15 ml, 0–0.35 M NaCl in 85 ml, 0.35–0.65 M NaCl in 15 ml, 0.65–1 M NaCl in 15 ml.

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DEAE Media Comparison

Comparing Purifications of Different DEAE Supports

Sample separation with the Macro-Prep DEAE support and two other commercially available DEAE supports. A 100 µl sample containing 9 mg/ml each of myoglobin, conalbumin, ovalbumin, BSA, and soybean trypsin inhibitor was loaded onto a 0.5 cm ID column containing 2 ml of support. Sample was loaded onto the column in 50 mM Tris-HCl, pH 7.6 (buffer A) and was eluted with 1 M NaCl in buffer A. The flow rate was 153 cm/hr (0.5 ml/min). Scales offset for comparison.

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Macro-Prep DEAE weak anion exchange media were developed using a macroporous methacrylate polymer and meet the demands of analytical, semipreparative, and process-scale applications.

This rigid, macroporous, hydrophilic 50 µm support provides exceptional capacity, resolution, and throughput in a chemically and mechanically stable form. Macro-Prep DEAE media operates well at low and medium pressures and exhibits minimal volume changes in response to variations in pH or ionic strength. Macro-Prep DEAE media provides consistent and reproducible results. Advantages include:

High capacity for biomolecules
High resolution of complex biological mixtures
Rigid polymer matrix that allows high flow rates at moderate pressures
Hydrophilic matrix that reduces nonspecific binding
Macroporous beads that enhance access to ionic sites
High chemical, mechanical, and thermal stability

Regulatory support files are available upon request. Bio-Rad Laboratories is an ISO 9001-registered corporation.