Detergent Removal

 

Detergents are widely used in solubilization of membrane and viral envelope proteins, viral removal/inactivation and vaccine purification. These detergents must eventually be removed downstream in the process after use.

 

Removal techniques such as gel filtration, size exclusion chromatography or dialysis have been used extensively in the biopharmaceutical industry, but they suffer from disadvantages such as dilution of the protein, and have limited effectiveness when dealing with detergents with low critical micelle concencentration (CMC). Therefore, a detergent with a low CMC like Triton X-100 requires different removal techniques. Triton X-100 is a non-ionic detergent with an approximate molecular weight 625 and CMC value of 0.24 mM. Its structure is shown below:

 

 

 

AMBERCHROM™ CG161 and AMBERCHROM™ CG71 chromatographic resins have been used in commercial purification processes for several decades and both exhibit excellent detergent capacities for non-ionic, zwitterionic, and anionic detergents. As demonstrated in tables 1 and 2 below, they are extremely effective for removing detergents from bioprocessing feed streams while providing high protein recoveries.

 

 

Table 1: Detergent Capacities
AMBERCHROM™ Resins	Detergent	1% Capacity g/L
Amberchrom™ CG161	Triton X-100	158
Amberchrom™ CG71	Triton X-100	91
Amberchrom™ CG161	Tween 20	100
Amberchrom™ CG71	Tween 20	85
Amberchrom™ CG161	CHAPS	25
Amberchrom™ CG71	CHAPS	91
Amberchrom™ CG161	SDS	131
Amberchrom™ CG71	SDS	75
Conditions: Column: 1.0cm ID x 7.0cm L - Flow Rate: 2 mL/min Buffer: 25mM sodium phosphate, pH 7.0 Detection: UV @ 280nm –Triton X-100 - UV @ 220nm –Tween20 and CHAPS RI –SDS

 

Table 2: Protein Capacities
Sample	MW	Amberchrom™ CG61 1% Capacity (g/L)	Amberchrom™ CG71 1% Capacity (g/L)
BSA	67,000	3	2
Ovalbumin	43,000	5	2
Lysozyme	14,000	65	20
Conditions: Column: 1.0cm ID x 7.0cm L - Buffer: 25mM sodium phosphate, pH 7.0 Flow Rate: 2mL/minute - Detection: UV @ 280nm

 

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