SP Strong Cation Exchange Membrane Chromatography

Introduction to CM Weak Ion-Exchange Membrane Chromatography Products

 

1. Overview

CM weak cation-exchange chromatography is a purification technology in which carboxymethyl groups are bonded onto the membrane to form a functional module. Separation is achieved based on the differences in the charge properties and charge densities of various biomolecules. Since most biological macromolecules contain carboxyl or hydroxyl groups, their charge characteristics and magnitude can be adjusted by changing the pH value of the buffer solution. After the biomolecules bind to the membrane module carrying opposite charges, elution is performed by altering the ionic strength or pH of the mobile phase. Molecules with weaker binding affinity are eluted first, while those with stronger binding affinity are eluted later, thereby achieving effective separation.

 

2. Product Advantages

2.1 Fast and efficient: High binding capacity can be achieved at flow rates up to 40 times faster than conventional resin-based chromatography. Compared with traditional packed-bed chromatography, membrane chromatography can shorten process time by 30–40 times. The typical operating flow rate is 10 MV/min.

2.2 High binding efficiency: Membrane chromatography demonstrates high loading capacity and high flow rates under low pressure drop conditions, enabling charged biomolecules to be captured in a single pass through the module.

2.3 Scalable and flexible: The complete range of membrane chromatography products can meet diverse biomacromolecule processing needs, covering stages from process development to large-scale manufacturing. The capsule design allows for single-use applications or can be cleaned and reused as required.

2.4 Increased productivity: The compact design minimizes facility footprint. By eliminating column packing, cleaning, cleaning validation, and column storage steps, processing can begin after equilibration with a relatively small volume of buffer. With no need for column packing, cleaning, or storage, labor costs can be reduced by up to more than 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
Membrane housing	Polypropylene
O ring	Silicone rubber

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 pressure differential	3bar(25℃)
Storage conditions	20% ethanol aqueous solution

Compared with conventional media, the service life of CM membrane chromatography is comparable to that of CM agarose gel 6FF.

Figure 1. Changes in the loading capacity of CM membrane chromatography after multiple uses in lysozyme testing.

Additionally, we evaluated the removal efficiency of host cell proteins and nucleic acids by membrane chromatography. The results are shown below.

Table 1. Removal rates of DNA and host cell proteins in CHO-expressed IgG material using CM membrane chromatography

A series of experiments demonstrated that CM membrane chromatography can effectively remove contaminants while maintaining a high recovery rate of target IgG

	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 Q Membrane	After Q Membrane	Log	Before Q Membrane	After Q Membrane	Log
1	96.7	423	6.9	1.79	7	6.1	0.06
2	97.4	438	5.8	1.88	7	4.8	0.16
3	94.7	513	9.6	1.73	6	3.6	0.22
4	95	32	4.6	0.84	6	4.7	0.11
5	96.3	45	4.6	0.99	8	5.1	0.20
6	96.5	158	8.2	1.28	8	4.6	0.24
7	96.4	267	6.5	1.61	9	6.8	0.12
8	96.8	298	5	1.78	9	7.4	0.09
9	97.1	746	5.6	2.12	4	3.5	0.06
10	96.6	39	5	0.89	4	3.9	0.01

Using Gudiling CM membrane chromatography in comparison with other brands, the loading capacity data are shown below.

Loading capacity(mg/mL)	Jingbiao 1	Guidling
Lysozyme	18	23
Trypsin	17	20

Table 2. Loading performance of different proteins in our product compared with competitor products

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

Figure 2. Loading capacity test of the CM membrane chromatography module using lysozyme as a standard protein.

The dynamic loading capacity was also compared with that of imported products. Verification showed that lysozyme could be eluted using 160 mM NaCl, which allows efficient capture of most target proteins.

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

 

4. Typical application cases

• Removal of DNA, viruses, and host cell proteins
• Capture of plasmids, viruses, and proteins, as well as purification of oligonucleotides
• Decolorization of yeast fermentation broth and high-capacity protein capture

 

5. Operating procedure / Workflow

5.1：Equipment preparation and assembly:

5.1.1： Install the membrane chromatography module on an AKTA chromatography system The installation method is similar to packed-bed chromatography media; ensure that the flow direction follows the arrow marked on the module. The module can be connected using Luer connectors or clamp connectors.

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

5.2.：Pre-use preparation

5.2.1：Set the inlet flow rate at 5–10 MV/min and perform pretreatment using 0.5 M NaOH for more than 5 MV to ensure the membrane reaches equilibrium.

5.2.2：Under the same flow rate conditions, perform pre-treatment with 1 M NaCl for more than 5 MV to ensure the membrane reaches equilibrium.

5.3. Chromatography process

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

5.3.2 After 0.22 μm pre-filtration, load the sample onto the column and continue until the entire sample has been applied or the chromatography loading capacity is reached.

5.3.3Wash with equilibration buffer for more than 10 MV until the UV value decreases to the baseline.

5.3.4 According to the process design, apply gradient elution or a linear elution system, and collect fractions in segments as required.

5.4 CIP Post-use treatment – CIP (Clean-in-Place) for membrane chromatography system.

5.4.1 Set the inlet flow rate to 5–10 MV/min and treat with 1 M NaOH for more than 10 MV until the UV value drops below the baseline.

5.4.2 After 30 minutes of circulating washing, rinse with water until the pH is between 7 and 8, then switch to 20% ethanol and continue cleaning until the conductivity remains nearly unchanged.

5.5Membrane 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 a solution containing 20% ethanol. The 20% ethanol solution should be inspected and replaced regularly.

 

6,Order information

CM-type weak cation exchange membrane chromatography capsule filter

	Laboratory scale	Small scale	Pilot scale	Production scale
Model	IEXCM0002ES	IEXCM0050ES	IEXCM0400ES	IEXCM5000ES
Membrane area	0.2ml	5ml	400ml	5L

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