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A novel method for poly(vinylalcohol)-based macroporous matrix (MP-PVA) preparation in aqueous media without usage of an outer porogens has been elaborated [1]. The method includes selective PVA oxidative cleavage at 1,2-diol fragments and self-polycondensation (self-crosslinking) of generated telechelic oligomers followed by crosslinking with an excess of glutaraldehyde (GA) (scheme 1). MP-PVA represents a highly crosslinked matix that is non-swellable in water and common organic solvents and has a high pore volume and pore surface and contains reactive aldehyde groups. Surface topography of MP-PVA has revealed its heterogeneous globule-like structure with open macropores of about 100–300 nm in diameter and large pores of about several microns. Dynamic light scattering study has elucidated the phase separation during the self-crosslinking step as the main factor responsible for macroporous structure generation. This two-phase structure of polymer gel is fixed by further crosslinking with GA. The prepared matrix has been successively applied as the parent platform for boronate affinity separation and quantification of glycated and non-glycated haemoglobin in human blood of diabetics patients. The reactive aldehyde-groups containing MP-PVA matrix may be promising for design of new macroporous sorbents and monoliths for affinity ligands immobilization and separation of biological macromolecules.