ABSTRACT
The advantages of the complex of recombinant human lactoferrin (rhLF) with europium ions have been used to establish quantitative parameters of specific interaction of rhLF with immobilized heparin-protein conjugate as a model of cell-surface heparan sulfate proteoglycans. Heparin coupled through terminal formyl by reductive amination to an inert protein was adsorbed through the protein part in the wells of a polystyrene microplate. The rhLF-Eu3+ complex obtained from native rhLF contains 0.8 mol of lanthanide ion per mol of protein (40 % saturation level). Equilibrium in the heterophase binding system is established within 1 min at room temperature, and the calculated association constant of the rhLF-heparin complex is 2.1. 107 M-1. The reversible and saturable character of binding rhLF labeled by Eu3+ at the active site to heparin was confirmed by the transition of rhLF-Eu3+ into the liquid phase when a 1000-fold molar excess of unlabeled rhLF was added to the system. Based on the affinity of rhLF for glycosaminoglycan, a blocking effect of this protein on the binding of the SARS-CoV-2 virus to the immobilized heparin-protein conjugate that imitates proteoglycan on the host cell surface was revealed. Pretreatment of the adsorbed conjugate with a solution of rhLF (10 mu g per well) reduces the specific binding of 100 ng of viral particles added to the well by approximately 80 %. The presented results allow one, in particular, to evaluate the integrity of the structure and activity of rhLF as a possible substance in food supplements and pharmaceuticals and may be useful in developing combined drugs for corona virus infection.