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Affinity Chromatography

Affinity chromatography (AFC) is performed on a support that is functionalized with a ligand that shows biological affinity for a particular enzyme or other biological molecule.

Almost all biomolecules can be purified on the basis of specific interaction between their chemical or biological structure and a suitable affinity ligand. Typical molecular pairs are antigens and antibodies, enzymes and coenzymes, and sugars with lectins. Thus, affinity chromatography distinguishes itself from, e.g., reversed phase and ion exchange chromatography, in that a highly specific interaction with the protein of interest is the cause for separation or purification.

Although affinity chromatography is not specific, in that no enzyme interacts with only one substrate, it is the most selective method for separating proteins. The selectivity is often based on spatial recognition via a 'lock-and-key' mechanism, as in antibody-antigen interactions.

Less selective general ligands have affinity for a whole class of proteins, which often require other chromatographic methods for purification of individual members. The term bioselective adsorption has been proposed as an alternative name for affinity chromatography.

Affinity Chromatography (AFC) offers the greatest potential specificity and selectivity for the isolation or purification of biomolecules. To enhance this advantage, three group specific ligands and one chemically active functionality are available through Tosoh Bioscience.

All analytical TSKgel AFC columns are based on the well-established 10 micron rigid TSKgel G5000PW resin. This resin features 100 nm pores that have an estimated exclusion limit of < 10 million Da along with excellent stability from pH 2 to 9.

Affinity Chromatography

Applications: Chelates with Zn⁺², Cu⁺² and Ni⁺² ions to target histidine residues on proteins

Literature:

• Catalog

• Affinity Column Products Table

• Instruction Manual

Particle Size (mean):	10 µm and 13 µm
Pore Size (mean):	100 nm
Functional Group:	iminodiacetic acid
pH Range:	2.0 - 12.0
Exclusion Limit (base resin, est.):	< 10,000,000 Da globular proteins
Ligand Concentration:	20 mmol/L resin

P/N	Description	Particle size	Housing material	ID (mm)	Length (cm)
0014440	TSKgel Chelate-5PW	10 µm	Glass	5	5
0014441	TSKgel Chelate-5PW	10 µm	Glass	8	7.5
0014442	TSKgel Guardgel kit for P/N 001441		Glass
0008645	TSKgel Chelate-5PW	10 µm	Stainless Steel	7.5	7.5
0008647	TSKgel Guardgel kit for P/N 0008645		Stainless Steel
0008646	TSKgel Chelate-5PW	13 µm	Stainless Steel	21.5	15
0020022	TSKgel BioAssist Chelate	10 µm	PEEK	7.8	5

Applications: Cis-diol groups found on carbohydates, glycosolyated proteins, catecholamines.

Literature:

• Catalog

• Affinity Column Products Table

• Instruction Manual

Particle Size (mean):	10 µm
Pore Size (mean):	100 nm
Functional Group:	m-aminophenyl boronate
pH Range:	2.0 - 9.0
Exclusion Limit (base resin, est.):	< 10,000,000 Da globular proteins
Adsorption Capacity:	40 mmol/L resin (sorbitol)

P/N	Description	Particle size	Housing material	ID (mm)	Length (cm)
0014449	TSKgel Boronate-5PW	10 µm	Glass	5	5
0014450	TSKgel Boronate-5PW	10 µm	Glass	8	7.5
0014451	TSKgel Guardgel kit for P/N 0014450		Glass
0013066	TSKgel Boronate-5PW	10 µm	Stainless Steel	7.5	7.5
0013125	TSKgel Guardgel kit for P/N 0013066		Stainless Steel

Applications: Blood clotting factors, glycoproteins, endoglycosidase, hyaluronidase, lipases, growth factors, nucleic acid-binding proteins

Literature :

• Catalog

• Affinity Column Products Table

• Instruction Manual

Particle Size (mean):	10 µm
Pore Size (mean):	100 nm
Functional Group:	heparin
pH Range:	5.5 - 9.0
Exclusion Limit (base resin, est.):	< 10,000,000 Da globular proteins
Adsorption Capacity for Antithrombin III:	< 1.8 g/L resin

P/N	Description	Particle size	Housing material	ID (mm)	Length (cm)
0014443	TSKgel Heparin-5PW	10 µm	Glass	5	5
0014444	TSKgel Heparin-5PW	10 µm	Glass	8	7.5
001445	TSKgel Guardgel kit for P/N 0014444		Glass
0013064	TSKgel Heparin-5PW	10 µm	Stainless Steel	7.5	7.5
0013121	TSKgel Guardgel kit for P/N 0013064		Stainless Steel

Applications: Reacts with amino or thiol groups, used primarily for attachment of specific antigens for antibody analysis

Literature :

• Catalog

• Affinity Column Products Table

• Instruction Manual

Particle Size (mean):	10 µm
Pore Size (mean):	100 nm
Active Group:	tresyl
pH Range:	2.0 - 12.0
Exclusion Limit (base resin, est.):	< 10,000,000 Da globular proteins
Ligand Concentration:	ca. 20 µmol/mL resin
Adsorption Capacity:	> 60 mg/g dry resin (coupling capacity / soybean trypsin inhibitor)

P/N	Description	Particle size	Housing material	ID (mm)	Length (cm)
0014455	TSKgel Tresyl-5PW	10 µm	Stainless Steel	6	4
0014456	TSKgel Tresyl-5PW	10 µm	Stainless Steel	7.5	7.5
0014457	TSKgel Tresyl-5PW	10 µm	Glass	5	5
0014458	TSKgel Tresyl-5PW	10 µm	Glass	8	7.5
0016208	TSKgel Tresyl-5PW Guardgel, 2g	10 µm