DEAE Weak Anion Exchange Membrane Chromatography

DEAE Weak Anion Exchange Membrane Chromatography – Product Introduction

 

1. Overview

DEAE weak anion exchange chromatography is a purification technique based on differences in the charge properties and charge density of various biomolecules. Since most biomacromolecules contain functional groups such as carboxyl or hydroxyl groups, their charge characteristics and magnitude can be adjusted by modifying the pH of the buffer solution. After biomolecules bind to an oppositely charged anion or cation exchange medium, separation is achieved by changing the ionic strength or pH of the mobile phase. Molecules with weaker binding affinity are eluted first, followed by those with stronger binding affinity, thereby accomplishing effective separation

 

2. Product Advantages

2.1 High efficiency: Achieves strong binding at flow rates up to 40 times higher than resin-based chromatography. Compared with traditional packed-bed chromatography, membrane chromatography shortens overall process time by approximately 30–40 times.

2.2 High binding efficiency: Membrane chromatography exhibits high binding capacity and high flow rate under low pressure drop, allowing charged biomolecules to be captured in a single pass through the column.

2.3 Scalable and flexible: The full series of membrane chromatography products can meet diverse biomacromolecule purification needs, covering all stages from process development to large-scale production. The capsule-type structural design supports both single-use operation and reuse after cleaning.

2.4 Improved productivity: The compact design minimizes facility footprint. By eliminating column packing, cleaning, cleaning validation, and column storage procedures, the system can be operated directly after buffer equilibration without column packing or storage. Labor costs can be reduced by up to 50%.

 

3. Technical Parameters

3.1 Structural Materials

	laboratory scale	small scale	pilot scale	production scale
Membrane volume	0.2ml	5ml	140ml	5L
Membrane support structure	Polypropylene (PP)
Membrane housing	Polypropylene (PP)
O ring	Silicone

3.2 Operating characteristics

	laboratory scale	small scale	pilot scale	Production scale
Membrane volume	0.2ml	5ml	400ml	5L
Recommended flow rate	1-6ml/min	25-150ml/min	2-12L/min	25-150L/min
Maximum operating temperature	35℃
Maximum operating pressure	3bar(25℃)
Maximum differential pressure	3bar(25℃)
Storage conditions	20%乙醇水溶液 20% ethanol aqueous solution

Figure 1. Changes in membrane chromatography loading capacity after multiple uses monitored with BSA.

Additionally, we evaluated the removal efficiency of host cell proteins and nucleic acids using DEAE membrane chromatography. The results are as follows.

	DNA				HCP
	IgG Recovery	Content(pg/mg of IgG)by RT PCR		Removal Factor	Content(ng/mg of IgG)by Elisa		Removal Factor
Run	%	Before DEAE Membrane	After DEAE Membrane	Log	Before DEAE Membrane	After DEAE Membrane	Log
1	96.7	423	3.5	2.08	7.8	5	0.19
2	97.4	438	6	1.86	7.7	4.9	0.20
3	94.7	513	8	1.81	6.3	1.9	0.52
4	95	32	2	1.20	6	4.2	0.15
5	96.3	45	6	0.88	8.1	5.2	0.19
6	96.5	158	3	1.72	8.7	6.2	0.15
7	96.4	267	2	2.13	9.8	7.1	0.14
8	96.8	298	7	1.63	9.4	8.2	0.06
9	97.1	746	5	2.17	4.3	2.6	0.22
10	96.6	39	2	1.29	4.4	1.7	0.41

Table 1. Removal rates of DNA and host cell proteins from CHO-expressed IgG using DEAE membrane chromatography.

A series of experiments demonstrated that Q membrane chromatography can effectively remove impurities, while maintaining a high recovery rate of the target IgG.

In addition, we compared Gudiling membrane chromatography with imported brands, and the loading capacity data are as follows.

Figure 2. Loading performance of different proteins on our membrane chromatography and competitor products

Overall evaluation shows that our loading capacity is comparable to that of imported products.

Figure 3. Performance of DEAE membrane chromatography module in collagen protein testing

Using the DEAE membrane chromatography module, validation results showed that most target collagen proteins could be captured and eluted using 135 mM NaCl.

Through testing under different elution conditions, we found that membrane chromatography and agarose gel chromatography exhibit similar elution behavior, where protein purity varies significantly under different salt concentrations. In practical R&D and production, it is necessary to quantitatively determine the optimal equilibrium elution conditions to obtain high-purity target proteins.

 

4. Classic application cases

Removal of DNA, viruses, host cell proteins, and endotoxins

Capture of plasmids, viruses, nucleic acids, and proteins, as well as purification of oligonucleotides

 

5. Operating workflow

5.1：Equipment preparation and assembly

5.1.1 The membrane chromatography module should be installed on the AKTA chromatography system in a manner similar to packed resin columns, ensuring that the flow direction aligns with the arrows and inlet direction. Connect using Luer connectors or clamp fittings.

5.1.2 Set the inlet flow rate to 5–10 MV/min and use equilibration buffer to purge air. Continue flushing until no bubbles are observed at the outlet, then connect the permeate outlet to the chromatography system

5.2：Pre-use treatment

5.2.1： Set the inlet flow rate to 5–10 MV/min and perform pre-treatment with 0.5 M NaOH for more than 5 MV to ensure that the membrane reaches equilibrium.

5.2.2： Under the same flow rate, further pre-treat with equilibration buffer (1× PBS) for more than 5 MV to ensure that the membrane reaches equilibrium.

5.3 Chromatography process

5.3.1

Set the inlet flow rate to 5–10 MV/min and pre-treat with equilibration buffer for more than 5 MV until the membrane reaches equilibrium.

5.3.2

After the sample is pre-filtered through a 0.22 μm filter, load the sample until loading is completed or the chromatography loading capacity is reached.

5.3.3

Wash with equilibration buffer for more than 10 MV until the UV absorbance decreases to the baseline level.

5.3.4

Use gradient elution or linear elution according to the process design, and collect samples in fractions as required.

5.4,CIP Post-use treatment – CIP of membrane chromatography device

5.4.1

Set the inlet flow rate to 5–10 MV/min and perform cleaning-in-place (CIP) treatment with 0.5 M NaOH for more than 10 MV until the UV absorbance drops below the baseline.

5.4.2

After circulating washing for 30 minutes, switch to water washing until the pH is between 7–8, then continue cleaning with 20% ethanol until the conductivity remains nearly constant.

5.5,Membrane chromatography storage:

After use and completion of CIP, the membrane module can be removed and stored by soaking in 20% ethanol, or stored online at room temperature in 20% ethanol solution. The 20% ethanol solution should be inspected and replaced periodically.

 

6.Ordering information

DEAE-type weak anion exchange membrane chromatography capsule filter

	Laboratory-scale	small scale	Pilot scale	Production scale
Product model	IEXD0002ES	IEXD0050ES	IEXD0400ES	IEXD5000ES
Membrane volume	0.2ml	5ml	400ml	5L

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