Selection of an effective affinity sorbent for the isolation of proteins with sialidase activity from human blood serum
Abstract
The aim of the study was to determine an effective affinity sorbent for isolating proteins with sialidase activity from human blood serum.
Materials and methods. Four affinity sorbents were used to obtain proteins with sialidase activity: 1) Neu5Ac(α2-8)Neu5Ac(α2-8)Neu5Ac trisialic acid immobilized on agarose, 2) sialic acid derivative (Neu5Acα)3-C3-PAA-biot immobilized on agarose, 3) sialic acid derivative immobilized on agarose 5- amino-Neu2en, 4) agarose-immobilized sialic acid derivative 4-amino-Neu5Ac2en. Isolation of the proteins responsible for LDL desiallation was performed using gradient polyacrylamide gel electrophoresis.
Results. The sorbent with (Neu5Acα)3-C3-PAA-biot most effectively binds proteins with neuraminidase activity. During electrophoresis of the obtained eluates, proteins with a molecular weight of 65 and 116 kDa are isolated.
