An Insight to a Highly Versatile Technique
29th October 2020 | 3pm GMT
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Phenyl Boronate affinity chromatography is a long-established useful tool for the specific capture and effective enrichment of target cis-diol-containing molecules, such as carbohydrates, glycoproteins, enzymes, RNA, nucleotides and nucleic acids. The webinar discusses the potential of Phenyl Boronate affinity chromatography as an additional alternative multi-modal tool for purification and polishing steps for different biopharmaceuticals.
The boronate ligand forms a pair of covalent bonds with molecules containing cis diols via a reversible esterification reaction. In acidic solutions, boronic acids adopt a trigonal planar form which can revert to a tetrahedral boronate anion upon hydroxylation in alkaline conditions. Both the acid and its conjugate base can bind to diol compounds. However, since the equilibrium constant for the tetrahedral is usually higher than that of the trigonal form, complexes are less stable in acidic conditions. This esterification is stronger if the two hydroxyl groups of the diol are on adjacent carbon atoms and in an approximately coplanar configuration, such as 1,2-cis-diol.
Phenyl Boronate ligand is an aromatic molecule and therefore, it can establish hydrophobic and π– π interactions. Other secondary ionic interactions between boronate ligand and proteins are possible through electrostatic attraction or repulsion effects, hydrogen bonding by the hydroxyl groups and charge transfer interactions. These are more likely to occur in acidic conditions, since in the trigonal uncharged form, the boron atom has an empty orbital and serves as an electron receptor for a coordination interaction, enabling Lewis acid–base interactions to occur.