Size Exclusion Chromatography

 Structure:

Size Exclusion Chromatography Resins

Size exclusion chromatography (SEC), also referred to as gel-filtration chromatography, uses porous particles to separate biological molecules of different sizes. Molecules that are smaller than the pore size can enter the particles and therefore have a longer path length and longer residence time than larger molecules that cannot enter the particles.

Proteins larger than the pore size can not enter the pores and will thus co-elute as the first peak in the chromatogram. These molecules are said to be totally excluded from the pores. Proteins that can at least partially enter the pores will have varying residence times dependent on the formula weight and hydrodynamic radius of each individual protein. Different proteins will therefore have different residence times when passing through the column. Proteins that elute in this portion of a chromatogram are said to be selectively permeable to the pores. Molecules that are smaller than the pore size are able to enter all of the pores in the resin and will therefore have the longest residence times. These molecules typically elute together as the last peak in a chromatogram and are said to be totally permeable to the pores. 

The main application of size exclusion chromatography is the fractionation and purification of synthetic and biological polymers, such as proteins, polysaccharides and nucleic acids. 

The advantages of this method include good separation of large molecules from the small molecules while preserving the biological activity of the particles to be separated. The technique is generally combined with others that further separate molecules by other characteristics, such as pI, charge, and affinity for certain compounds and is typically used as a polishing step. 

 
 
Calibration curves for globular proteins on
TOYOPEARL HW resins

 
Typical resolution changes based on
particle size




Literature :

 • Catalog

 • Instruction Manual

TOYOPEARL HW-40 resin is a hydroxylated methacrylic polymer for size exclusion chromatography. It is typically used for high throughput desalting and buffer exchange. It also can remove surfactants such as Triton X100 from protein solutions. It is often used for smaller biomolecules with exclusion limits up to 10,000 Da.

TOYOPEARL HW-40 resin is available in three particle sizes.  

Pore size (mean) 5 nm
Exclusion limit 10,000 Da
Particle size (mean)   75 µm (C-grade) 
45 µm (F-grade)
30 µm (S-grade)
Pressure rating 0.3 MPa
Shipping buffer 20% ethanol
pH stability 2 - 13

P/N Description Resin volume Particle size
  0007450 Toyopearl HW-40EC 500 mL 100 - 300 µm
  0007449 Toyopearl HW-40C 500 mL 50 - 100 µm
  0014683 Toyopearl HW-40C 1 L 50 - 100 µm
  0007969 Toyopearl HW-40C 5 L 50 - 100 µm
  0007448 Toyopearl HW-40F 500 mL 30 - 60 µm
  0014682 Toyopearl HW-40F 1 L 30 - 60 µm
  0007968 Toyopearl HW-40F 5 L 30 - 60 µm
  0007447 Toyopearl HW-40S 500 mL 20 - 40 µm
  0014681 Toyopearl HW-40S 1 L 20 - 40 µm
  0007967 Toyopearl HW-40S 5 L 20 - 40 µm

TOYOPEARL HW-50 resin is a hydroxylated methacrylic polymer for size exclusion chromatography of proteins between 500-80,000 Da.

TOYOPEARL HW-50 resin is available in two particle sizes.

Pore size (mean) 12.5 nm
Exclusion limit 80,000 Da
Particle size (mean)   45 µm (F-grade) 
30 µm (S-grade)
Pressure rating 0.3 MPa
Shipping buffer 20% ethanol
pH stability 2 - 13

P/N Description Resin volume Particle size
  0007453 Toyopearl HW-50F 500 mL 30 - 60 µm
  0014685 Toyopearl HW-50F 1 L 30 - 60 µm
  0008060 Toyopearl HW-50F 5 L 30 - 60 µm
  0007452 Toyopearl HW-50S 500 mL 20 - 40 µm
  0014684 Toyopearl HW-50S 1 L 20 - 40 µm
  0008059 Toyopearl HW-50S 5 L 20 - 40 µm

TOYOPEARL HW-55 resin is a hydroxylated methacrylic polymer bead for size exclusion chromatography of proteins between 1,000-700,000 Da. It also is the base bead for all TOYOPEARL "-550" designated products.

TOYOPEARL HW-55 resin is available in two particle sizes.

Pore size (mean) 50 nm
Exclusion limit  700,000 Da
Particle size (mean)   45 µm (F-grade) 
30 µm (S-grade)
Pressure rating 0.3 MPa
Shipping buffer 20% ethanol
pH stability 2 - 13

P/N Description Resin volume Particle size
  0007457 Toyopearl HW-55F 500 mL 30 - 60 µm
  0014687 Toyopearl HW-55F 1 L 30 - 60 µm
  0008063 Toyopearl HW-55F 5 L 30 - 60 µm
  0007456 Toyopearl HW-55S 500 mL 20 - 40 µm
  0014686 Toyopearl HW-55S 1 L 20 - 40 µm
  0008062 Toyopearl HW-55S 5 L 20 - 40 µm

TOYOPEARL HW-65 resin is a hydroxylated methacrylic polymer bead for size exclusion chromatography of proteins between 40,000-5,000,000 Da. It also is the base bead for all TOYOPEARL "-650" designated products.

TOYOPEARL HW-65 SEC resin is available in two particle sizes. 

Pore size (mean) 100 nm
Exclusion limit 5,000,000 Da
Particle size (mean)   45 µm (F-grade) 
30 µm (S-grade)
Pressure rating 0.3 MPa
Shipping buffer 20% ethanol
pH stability 2 - 13

P/N Description Resin volume Particle size
  0007465 Toyopearl HW-65F 500 mL 30 - 60 µm
  0014689 Toyopearl HW-65F 1 L 30 - 60 µm
  0008069 Toyopearl HW-65F 5 L 30 - 60 µm
  0007464 Toyopearl HW-65S 500 mL 20 - 40 µm
  0014688 Toyopearl HW-65S 1 L 20 - 40 µm
  0008068 Toyopearl HW-65S 5 L 20 - 40 µm

TOYOPEARL HW-75 resin is a hydroxylated methacrylic polymer bead for size exclusion chromatography of proteins between 500,000-50,000,000 Da.

TOYOPEARL HW-75 resin is available in two particle sizes.

Pore size (mean) > 100 nm
Exclusion limit 50,000,000 Da
Particle size (mean)   45 µm (F-grade) 
30 µm (S-grade)
Pressure rating 0.3 MPa
Shipping buffer  20% ethanol
pH stability 2 - 13

P/N Description Resin volume Particle size
  0007469 Toyopearl HW-75F 500 mL 30 - 60 µm
  0014691 Toyopearl HW-75F 1 L 30 - 60 µm
  0008072 Toyopearl HW-75F 5 L 30 - 60 µm
  0007468 Toyopearl HW-75S 500 mL 20 - 40 µm
  0008071 Toyopearl HW-75S 5 L 20 - 40 µm