Место издания:FEBS Open Bio Prague, Czech Republic
Первая страница:475
Последняя страница:475
Аннотация:Surface modification for biomedical,
environmental, agricultural and security applications can be performed physically
and chemically by surface coverage or grafting with polymers and conjugating ligands. The immobilized substances must
retain their structures and not desorb from the functionalized surfaces. Hydrophobic, electrostatic and vanderWaals
interactions, coordination and covalent binding are involved in the adsorption. Our aim was the development of the
functionalization method for immobilization of native proteins and viable cells. Cationic polymers with NH2groups
were used for functionalization of charged and uncharged surfaces of planar slides, wells of plates and spherical
nanoparticles. Poly(allylamine)s (PAA) and polylysines (pLys) of different molecular weights formed interfaces of the
thicknesses ~1.52
nm for PAA 65 kDa as measured by: 1) the atomic force microscopy on mica slides; 2) the labelfree
biosensor with registration of the critical angle of total internal reflection on photonic crystal surface covered with SiO2.
Sorption capacity of 0.1 mg/ml PAA 65 kDa exceeded the values of other polyamines with different concentrations.
Physisorption of proteins on PAAlayer
is reversible and up to 70% of attached proteins can be removed. Glutaraldehyde
provides stable chemical crosslinking
of the compounds containing primary NH2groups
with the aminated surfaces. The
proteins immobilized on the pAAcovered
surface retained their ability to bind with specific monoclonal and polyclonal
antibodies. Bacterial cells attached to PAA65covered
surfaces could express the green fluorescent protein (GFP) gene
under control of the inducible lac promoter. Eukaryotic cells also remained alive on PAAtreated
slides as shown by
fluorescent staining. Thus, treatment of plain and spherical surfaces with PAA is convenient for attachment of
biopolymers and viable cells.
The authors acknowledge funding from the Russian Science Foundation [177530064
to D.V.K.].