Avidin/Biotin Affinity Supports

Ami.R. Gel is favoured in research applications for its efficiency and ease of use. Diagnostic applications also benefit from its high binding efficiency, capacity, and ease of use (especially in manufacturing). In pharmaceutical processing, Ami.R. Gel and Ami.R. Gel Fast Flow are also replacing conventional activated gels thanks to their extreme stability and cleanability

Literature

Grandics, P. et al. 1990 Ann. N.Y. Acad. Sci. 589:148-156

Thalley, B. and Carroll, S. 1990. Bio-Technology 8:934-938

Schoepfer, R. et al. 1990. Neuron 5:35-48

Buffers

Mild IgG elution buffer

neutral pH

UP38591A

Elution under neutral and non-denaturing conditions !

Preserves biological activity (>97%)
Increased yield (>97%)
Increases the longevity of immunoadsorbents
50-100 fold economy per purification cycle cost

Acidic elution not only leads to significant losses of antibody activity, but also causes subtile changes in the IgG conformation.

Chaotropic agents exhibit higher bioactivity but with lower yields.

Uptima Mild Elution Medium nearly delivers quantative recovery of antibodies with no detectable loss of bioactivity.

Technical Sheet

Recovery of bound IgG was carried out by using either Uptima mild Elution Medium, 1M acetic acid or chaotropic (3 M NH4SCN, pH 7.0) elution buffers. Then yield and activity was determined.

PBS

UP68723

Immobilized (Strept)avidins and biotins

Avidin and (Strept)avidin have considerable affinity for biotin, making the interaction almost irreversible as elution requires harsh condition (4M Urea or 8M Guanidin). This makes Avidin- Streptavidin- and Biotin- immobilized supports ideal for the preparation of affinity columns. For example, a biotinylated peptide or antibody can by irreversibly immobilized to further capture specific ligands (soluble molecule or particule) from complexe samples (a cell suspension, a bacterial lysate, or a biological fluid). At the other hand, immobilized biotins permit avidin capture.

Monomeric avidin, with it’s single binding valency, exhibits lower interaction with biotin than (tetrameric) avidin does. This permits to elute bound biotinylated molecules under relatively milder conditions, as 2-5mM biotin or with pH2.8

Imino-Biotin has a lower affinity for (strept)avidin products, allowing reversible binding for purification purposes.

Irreversible binding biotin/avidin supports

Great for the preparation of affinity supports !

Applications

Immobilization of biotinylated antibodies, peptides,antibodies, lectins, DNA or any other probe to create affinity supports for purification and immunoprecipitation
Removal of biotin and biotinylated molecules from complexe samples

Purification systems based on biotin/avidin

Ideal for the immunoprecipitation and purification of biotinylated ligands

Applications

Purification of biotinylated molecules, notably when sensitive to denaturation under chaotropic conditions
Biotinylated ligands (antibodies, peptide, receptor, DNA…) bound to their ligand in complexe mixtures will be captured and can be eluted specifically (purification or immunoprecipitation)

Literature

Buckie, J.W. and Cook, G.M.W.; Specific isolation of surface glycoproteins from intact cells by biotinylated concanavalin A and immobilized streptavidin; Anal. Biochem. 1986; 156; 463-472.

Gretch, D.R., Suter, M. and Stinski, M.F. ; The use of biotinylated monoclonal antibodies and streptavidin-affinity chromatography to isolate herpes virus hydrophobic proteins or glycoproteins. Anal. Biochem. 1987; 163; 270-277.

Mitchell, L.G. and Merril, C.R. ; Affinity generation of single-stranded DNA for dideoxy sequencing following the polymerase chain reaction ; Anal. Biochem. 1989; 178; 239-242.

Literature

Guchait R.B., Polakis S.E., Dimroth P., Stoll E., Moss J., and Lane M.D. ; Acetyl co-enzyme A system Escherichia coli : Purification and properties of the biotin carboxylase, carboxyltransferase, and carboxyl carrier protein components ; Biol.Chem.1974; 249; 6633-6645

Tucker J, Grisshammer R ; Purification of a rat neurotensin receptor expressed in Escherichia coli.; Biochem. J. 1996 Aug 1; 317( Pt 3); 891-899

Avidin Agarose	UP340909	2 ml	Mon. avidin - Agarose Kit	UP29337A	1 Kit
	UP34090A	5 ml
	UP34090B	10 ml
Avidin immobilized onto a 4 % modified agarose, supplied at 50 % suspension 50 µg of biotin / ml of gel Technical Sheet			Complete kit for the purification of biotinylated molecules Contains 2 ml of gel, 250 ml of regeneration buffer, 250 ml of elution buffer, 500 ml of washing buffer 10 µg of d-biotin per ml of wet gel Technical Sheet