Affiliations: Department of Physiology and Biophysics, Keck School of Medicine, University of Southern California, Los Angeles, CA 90033, USA
Note: [] Address for correspondence: Dr. Björn Neu, Department of Physiology and Biophysics, Keck School of Medicine, 1333 San Pablo Street, MMR 626, Los Angeles, CA 90033, USA. Tel.: +1 323 442 1267; Fax: +1 323 442 1617; E‐mail: [email protected].
Abstract: Recent studies have shown that the covalent attachment of poly(ethylene glycol), abbreviated as PEG, to the surface of human red blood cells (RBC) leads to masking of membrane antigenic sites and inhibition of RBC aggregation. The effects of PEG coating on the regions near the RBC glycocalyx were thus explored using cell micro‐electrophoresis. Both linear (3.35, 18.5, 35.0) and an 8‐arm 35.9 kDa reactive PEG were used; in one series, thick cross‐linked coats were obtained using a branched PEG amine as a cross‐linker. The results indicate marked decreases of RBC mobility (up to 90%) which were affected by polymer molecular mass and geometry. Since PEG is neutral and its covalent attachment is predominantly to primary amine groups, such decreases of mobility most likely reflect structural changes near and within the RBC glycocalyx rather than decreased surface charge density. Experimental data were analyzed using a theoretical approach which allows calculation of the thickness and friction of the polymer layers: (1) for linear PEGs, thickness increased and friction decreased with polymer mass; (2) compared to linear PEGs of similar molecular mass, thickness was less and friction was greater for the branched PEG; (3) cross‐linked PEG coatings were more than 50 nm thick and were insensitive to changes of ionic strength. These observations are consistent with the aggregation behavior of PEG‐coated RBC and indicate the usefulness of micro‐electrophoresis methods for studies of covalently‐attached polymers: the resulting calculated thickness and friction factors should be of value in achieving desired cellular surface characteristics or levels of cell–cell interaction.
Keywords: Micro‐electrophoresis, poly(ethylene glycol), red blood cell, polymer‐coating
Journal: Biorheology, vol. 40, no. 4, pp. 477-487, 2003
