Biospecific affinity chromatography

Sundberg, L., and Porath, J. (1974) Preparation of adsorbents for biospecific affinity chromatography. 

Figure 6.14 Schematic representation of the principle of biospecific affinity chromatography. The chosen affinity ligand is chemically attached to the support matrix (agarose bead) via a suitable spacer arm. Only those ligands in solution that exhibit biospecific affinity for the immobilized species will be retained...

It is not normally prudent to employ biospecific affinity chromatography as an initial purification step, as various enzymatic activities present in the crude fractions may modify or degrade the expensive affinity gels. However, it should be utilized as early as possible in the purification procedure in order to accrue the full benefit afforded by its high specificity. 

Sundberg, L., and Porath, J. (1974) Preparation of adsorbents for biospecific affinity chromatography. I. Attachment of group containing ligands to insoluble polymers by means of bufunctional oxiranes. /. Chromatogr. 90, 87—98. 

J. Porath and T. Kristiansen, Biospecific Affinity Chromatography and Related Methods, The Proteins Vol. I (H. Neurath and R. L. Hill, Eds.), Academic Press, New York, 1975, Pg 95-178. 

Immobilized metal affinity chromatography has been shown to be effective for isolating proteins from crude mixtures, as well as for selective separations of closely related proteins [2]. With respect to separation efficiency, IMAC compares well with biospecific affinity chromatography and the immobilized metalion complexes are much more robust than antibodies or enzymes. These factors make IMAC particularly well suited for scale-up to process scale chromatography. The main scale-up points to be aware of are the degree to which the column is metal saturated, the chelating agent content of the sample, and the potential of leached metal (or its interactions) within the product eluate. 

Yasukochi, Y. and B.S. Masters (1976). Some properties of a detergent-solubilized NADPH-cytochrome c (cytochrome P-450) reductase purified by biospecific affinity chromatography. J. Biol. Chem. 251, 5337-5344. 

For the separation biospecific affinity chromatography was used, where in addition to the adsorption and desorption zone a washing zone has to be included. Gottschlich and Kasche [22] used the same apparatus for the separation of monoclonal antibodies from a fermentation broth. 

Ziska P, Franz H, Kindt A (1978) The lectin from Viscum album L. Purification by biospecific affinity chromatography. Experentia 34 123-124... 

A new principle of biospecific affinity chromatography was demonstrated following the addition of a solution of hydroxocobalamin to the agarose cyclic imidocarbonate derivative (8A). The bond formed is non-covalent and temperature sensitive. 

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