Laserchrom HPLC - Shodex Affinity Chromatography Columns

Shodex Affinity Chromatography Columns


Affinity Chromatography is essentially a sample purification technique, used primarily for biological molecules such as proteins. Unlike other forms of HPLC, where most if not all components of the sample have an attraction to the column, in Affinity Chromatography the column contains a ligand which is specific to a sample component (or group of components) of interest. This component (or group of components) alone is bound to the column and everything else elutes unretained. Then by changing the eluent composition using a gradient, the adsorbed species (or group of species) is eluted. Column Specificity. Since columns have quite highly specific active sites, a wide variety of columns is required, even to separate groups of compounds. It is not normally commercially viable to produce columns for individual compounds. However there are many groups of compounds which can be isolated using affinity chromatography, and details of these is given on the table below. It should be noted that if a column is used to isolate a group of compounds from a mixture, they will usually be all retained and then eluted together. Process of Affinity Chromatography. First a column must be found containing a ligand which will specifically bind the sample component of interest. Then two sets of eluent conditions must be developed. The first is used to introduce the sample, allow it to bind to the ligand, and wash out all other components from the column. The second eluent is used to elute the sample from the column without denaturing it. It is normal to use a gradient, although a stepwise process can also be used. Group-Specific Columns. These are the most widely available, as per the list below. Each contains a rigid gel, onto which is bonded a spacer group to minimise steric hindrance when a larger molecule approaches, and onto that is bonded the ligand. Because the columns have an "on-off" retention sequence, the columns need only be short. 8mm diameter is often used to give a good loading capacity, although smaller bore columns are also available. .
Other features to note are:
Particle Size	Typically 6-20 μm.
Efficiency	Some columns give sharper peaks than others
Max Flow Rate	3 ml/min
Max Pressure	About 45psi, 0.3 MPa
pH range	2-10, depending on the ligand
Organic solvent	IPA or ethylene glycol, max 10%
More information is available below or at the Shodex website www.shodex.com

Column Details - Affinity Chromatography

Packing	Particle Size (μm)	Ligand	Applications:
AFpak AAB-894	18	Aminobenzamidine	Serine Protease etc
AAB columns contain aminobenzamidine (a serine protease inhibitor) at a loading of 100uMol/g. An example application for the analysis of trypsin can be found following the link to the Shodex website: For pricing and ordering information click here, or for more info see www.shodex.com
AFpak AAF-894	18	Acriflavine	RNA, DNA, Vitamins etc
AAF columns contain acriflavine at a loading of 10uMol/g, which is suitable for the analysis, fractionation and purification of heterocyclic materials such as nucleic acids. Acriflavine is a tricyclic derivative of acridine which has antifungal and antiseptic properties. The column loading capacity is about 1.8mg/column (8.0 x 50mm) ATP Na. Applications can be found on the Shodex website link for various nucleotides, and for Riboflavin (Vitamin B2): For pricing and ordering information click here, or for more info see www.shodex.com
AFpak AAM-894	18	5'AMP	NAD, ATP-dependent enzymes etc
AAM columns contain 5'ATP at a loading of 10uMol/g. Adenosine monophasphate is made up of adenine, ribose and a phosphate group, and is a nucleotide found in RNA. The loading capacity of this column is 1.5mg/g Lactic dehydronase. No applications are currently featured on the Shodex website, so if you have an application and would like to send it to us, we would be pleased to include it here! For pricing and ordering information click here, or for more info see www.shodex.com
AFpak AAP-894	18	Aprotinin	Serine Protease etc
AAP columns contain Aprotinin at a loading of 5mg/g. Aprotinin is a polypeptide (58 amino acids) extracted from cows lung tissue. It acts as an inhibitor for serine proteases, specifically trypsin, chymotrypsin and plasmin. The loading capacity of these columns is about 3mg/g of trypsin. For pricing and ordering information click here, or for more info see www.shodex.com
AFpak AAV-894	18	Avidin	Biotin, biotin derivatives
AAV columns contain Avidin at a loading of 5mg/g. Avidin is glycoprotein found in egg white, that has a very strong affinity for biotin. Used with biotin, it has a loading capacity of 8ug/column (8.0 x 50mm). An example application is given on the link to the Shodex website: For pricing and ordering information click here, or for more info see www.shodex.com
AFpak ABA-894	6	Bovine Serum Albumen	Chiral separation of amino acids, avidin etc
ABA columns contain BSA at a loading of 20mg/g. The column has applications for chiral HPLC as well as for Affinity chromatography. An example application is given on the link to the Shodex website: For pricing and ordering information click here, or for more info see www.shodex.com
AFpak ABT-894	18	Biotin	Avidin etc
ABA columns contain Biotin, otherwise known as vitamin B7. Biotin has a strong affinity for avidin etc, An example application is given on the link to the Shodex website: For pricing and ordering information click here, or for more info see www.shodex.com
AFpak ACA-894	18	Concanavalin A	Glycoproteins, polysaccharides etc
ACA columns contain Concanavalin, at a level of 15mg/g. Concanavalin A is a plant lectin with a polypeptide structure. Two example applications are given on the link to the Shodex website: For pricing and ordering information click here, or for more info see www.shodex.com
AFpak ACB-894	18	Cibacron blue	Albumin, NAD-dependent enzymes etc
ACB columns contain Cibacron blue, with a loading of 40uMol/g. Cibacron blue binds stongly to proteins such as albumen, and has a loading capacity of approx 20mg/column (8.0 x 50mm) An example application is given on the link to the Shodex website: For pricing and ordering information click here, or for more info see www.shodex.com
AFpak ADS-894	18	Dextran sulphate	Lipoproteins, blood coagulation factors etc
ADS columns contain Dextran sulphate at a loading of 30mg/g. It has a loading capacity of 5mg/g usin LDL. A couple of example applications are given on the link to the Shodex website: For pricing and ordering information click here, or for more info see www.shodex.com
AFpak AED-894	18	Ethylenediaminediacetic acid	Nucleic Acids, serum proteins etc
AED columns contain ethylenediamine diacetic acid at a level of 30uMol/g. This is quite a strong chelating agent, with affinity for a number of species. A couple of example applications are given on the link to the Shodex website: For pricing and ordering information click here, or for more info see www.shodex.com
AFpak AGA-894	6	N-Acetylglucosamine	Lectins, carbohydrate-metabolising enzymes etc
AGA columns contain N-acetylglucosamine. An example application is given on the link to the Shodex website: For pricing and ordering information click here, or for more info see www.shodex.com
AFpak AGE-894	18	Gelatin	Fibronectin etc
AGE columns contain gelatin at a level of 30mg/g. The loading capacity of the column is approx 150mg/g fibronectin. An example application is given on the link to the Shodex website: For pricing and ordering information click here, or for more info see www.shodex.com
AFpak AGT-894	18	Glutathione	Enzymes related to Glutathione such as glutathione reductase etc
AGT columns contain glutathione at a level of 50uMol/g. Glutathione is a tripeptide structure which acts as a cell antioxidant. An example application is given on the link to the Shodex website: For pricing and ordering information click here, or for more info see www.shodex.com
AFpak AHR-894	18	Heparin	Lipoproteins, Blood coagulation factors etc
AHR columns contain heparin at a level of 5mg/g. Heparin is a highly sulphonated glycosaminoglycan, used as an anticoagulant. The column loading capacity is approx. 4mg/ column (8.0 x 50mm) Lysozyme. A couple of example applications are given on the link to the Shodex website: For pricing and ordering information click here, or for more info see www.shodex.com
AFpak AIA-894	18	Iminodiacetic acid	Interferon, serum proteins etc
AIA columns contain iminodiacetic acid (IDA) at a level of 70uMol/g. IDA columns can be used alone, or in conjunction with copper ions for the isolation of proteins or ribonucleosides.The column loading capacity is approx. 70mg/ column (8.0 x 50mm) BSA. A couple of example applications are given on the link to the Shodex website: For pricing and ordering information click here, or for more info see www.shodex.com
AFpak ALC-894	18	Lentil Lectin	Glycoproteins, polysaccharides etc
ALC columns contain lentil lectin at a level of 6-7mg/g. Lectins are a type of receptor protein which specifically interacts with sugar molecules. For a similar but slightly different interaction, see the ACA column, listed above. An example application for the purification of fibrinogen is given on the link to the Shodex website: For pricing and ordering information click here, or for more info see www.shodex.com
AFpak ALS-894	10	Lysine (LCA)	Plasminogen, plasminogen activator, RNA, DNA etc
ALS columns contain Lysine. The loading capacity of the column is approx 54ug/g plasminogen. Lysine is one of the basic amino acids. An example application is given on the link to the Shodex website: For pricing and ordering information click here, or for more info see www.shodex.com
AFpak ANA-894	18	NAD	NAD-dependent dehudrogenase etc
Nicotinamide adenine dinucleotide (NAD+) is the oxidised form of NADH. ANA columns contain NAD at a level of 10uMol/g. The loading capacity of the column is approx 1.4mg/g lactic hydrogenase. An example application is given on the link to the Shodex website: For pricing and ordering information click here, or for more info see www.shodex.com
AFpak AOV-894	18	Ovomucoid	Trypsin-like protease etc
AOV columns contain ovomucoid at a level of 10mg/g. Ovomucoids are the glycoprotein protease inhibitors found in avian egg white. There are other protease inhibitors in ovomucoids as well. The loading capacity of the column is approx 100mg/g Trypsin . An example application is given on the link to the Shodex website: For pricing and ordering information click here, or for more info see www.shodex.com
AFpak APA-894	18	Protein A	Immunoglobulin G, immune complexes etc
APA columns contain Protein A at a level of 4mg/g. The loading capacity of the column is approx 20mg/g human IgG. Protein A is a large surface protein which is used extensively in biochemical research because of its ability to bind immunoglobulins. It binds with high affinity to human IgG1 and IgG2 as well as mouse IgG2a and IgG2b. Protein A binds with moderate affinity to human IgM, IgA and IgE as well as mouse IgG1. It does not react with human IgG3 or IgD, nor will it react to mouse IgG3, IgM, IgA or IgE. An example application is given on the link to the Shodex website: For pricing and ordering information click here, or for more info see www.shodex.com
AFpak APB-894	10	Aminophenylboronic Acid	Nucleic Acids, Catecholamines etc
APB columns contain aminophenylboronic acid at a level of 800uMol/g. The column loading capacity is approx. 0.2mg/ column (8.0 x 50mm) sorbitol. A couple of example applications are given on the link to the Shodex website: For pricing and ordering information click here, or for more info see www.shodex.com
AFpak APD-894	18	Procion Red	NAD, NADP-dependent enzymes, interferon etc
APD columns contain procion red at a level of 40-60uMol/g.The column loading capacity is approx. 72mg/ column (8.0 x 50mm) BSA. Procion Red (HE-3B) is a triazine dye which has an interaction with the catalytic centres of various hydrogenase enzymes. An example application is given on the link to the Shodex website: For pricing and ordering information click here, or for more info see www.shodex.com
AFpak APE-894	18	Phosphorylethanolamine	C-reactive proteins etc
APE columns contain phosphorylethanolamine. The column loading capacity is approx. 0.91mg/ g C-reactive protein. C-reactive protein (CRP) is a plasma protein, an acute phase protein produced by the liver. It is a member of the pentraxin family of proteins. It should not be confused with C-peptide or Protein C. The pentraxin family of proteins interact specifically with phosphorylethanolamine An example application is given on the link to the Shodex website: For pricing and ordering information click here, or for more info see www.shodex.com
AFpak APG-894	18	Protein G	Immunoglobulin G, immune complexes etc
APG columns contain Protein G at a level of 4-5mg/g. The column loading capacity is approx. 10mg/g human IgG. The IgG antibodies bind with different affinities to Protein A and Protein G. Protein G is recommended for mouse IgG1 and IgG3, goat IgG and hamster IgG. It will also bind sheep and rat IgG but only with a weak affinity. An example application is given on the link to the Shodex website: For pricing and ordering information click here, or for more info see www.shodex.com
AFpak APH-894	18	Phenylalanine	Alkalophilic Protease etc
APH columns contain phenylalanine at a level of 50uMol/g. The column loading capacity is approx. 9mg/g Subtilisin Carlsberg . For pricing and ordering information click here, or for more info see www.shodex.com
AFpak APR-894	18	Protamine	IgM etc
APR columns contain Protamine at a level of 4-5mg/g.The column loading capacity is approx. 1.9mg/ column (8.0 x 50mm) human IgM. Protamine (PRM) is a small nuclear protein found in sperm. There are two human protamines, PRM1 and PRM2, but most animals only have one. IgM is a very large molecular weight antibody, primarily responsible for the agglutination of red blood cells. A couple of example applications are given on the link to the Shodex website: For pricing and ordering information click here, or for more info see www.shodex.com
AFpak ARC-894	18	RCA-1	Glycoproteins, polysaccharides etc
ARC columns contain RCA-1 at a level of 20mg/g. RCA is ricinus communis agglutinin, and is a lectin, acting in a similar manner to Concanavalin A (ACA) or Lentil Lectin (ALC) above. An example application is given on the link to the Shodex website: For pricing and ordering information click here, or for more info see www.shodex.com
AFpak AST-894	18	Soybean Trypsin Inhibitor	Trypsin-like protease etc
AST columns contain soybean trypsin inhibitor at a level of 20mg/g. The column loading capacity is approx. 100mg/g trypsin. An example application is given on the link to the Shodex website: For pricing and ordering information click here, or for more info see www.shodex.com
AFpak AWG-894	18	Wheat germ agglutinin	Glycoproteins, polysaccharides etc
AWG columns contain wheat germ agglutinin (WGA) at a level of 14mg/g. WGA is a protein containing a group of closely related isolectins. WGA can bind oligosaccharides containing terminal N-acetylglucosamine or chitobiose, structures which are common to many serum and membrane glycoproteins. An example application is given on the link to the Shodex website: For pricing and ordering information click here, or for more info see www.shodex.com

Acknowledgement is given here to Wikipedia for some of the information used in the sections above.