Polar silica-based stationary phases. Part III- Neutral silica stationary phase with surface bound maltose for affinity chromatography at reduced non-specific interactions.

This research article reports the coating of large pore silica microparticles with a maltose layer to which bioaffinity ligands were attached via reductive amination reaction between the aldehyde activated maltose and the amino groups of the bioaffinity ligands. This was achieved first by the periodate oxidation of the maltose-silica (MALT-silica) yielding pairs of aldehyde groups at each monosaccharide ring. These di-aldehyde functionalities were then reacted with the primary amino groups of protein bio-affinity ligands and eventually formed Schiff bases (i.e., aldimines) which were reduced using the mild reducing agent sodium cyanoborohydride to form stable amine linkages between the immobilized protein ligands and the maltose layer. Anti-human serum albumin antibody (aHSA), anti-human serum transferrin antibody (aTf) and concanavalin A (Con A) were the bio-affinity ligands immobilized onto the MALT-silica and were evaluated in high performance affinity chromatography (HPAC), namely immunoaffinity chromatography (IAC) and lectin affinity chromatography (LAC). Our initial studies reported here revealed zero or reduced nonspecific interactions with the two immunoaffinity sorbents (i.e., aHSA-MALT-silica and aTf-MALT-silica) and the lectin affinity sorbent (i.e., Con A-MALT-silica). The absence of nonspecific interactions is attributed to the hydrophilicity of the maltose layer and its shielding effect of the residual silanols (i.e., unreacted silanols) on the silica surface. Conversely, the IAC and LAC sorbents exhibited specific interactions with the target biomolecules, namely human serum albumin (HSA) and transferrin (Tf) in the case of aHSA-MALT-silica and aTf-MALT-silica columns, respectively, and glycoproteins known for their affinity to Con A in the case of Con A-MALT-silica column.