IN-VITRO PROPERTIES AND ORGAN UPTAKE OF RAT BAND-3 CROSS-LINKED ERYTHROCYTES

Chemical conditions for cross-linking rat erythrocytes with bis(sulfos uccinimidyl)suberate (BS') and 3,3' dithiobis(sulfosuccinimidyl propio nate) (DTSSP) have been studied. These two cross-linking reagents seem to react with band 3 protein in rat erythrocyte membrane. Two differe nt conditions have been assayed using different cell/cross-linker conc entration ratios. Similar cell volumes were observed in cross-linked r at erythrocytes compared to rat control erythrocytes. Cell yields afte r cross-linking account for about 65% when a high ratio of cell-to-cro ss-linker was used. Slightly lower cell yields (about 62%) were obtain ed when this ratio was reduced. Estimation of band 3 cross-linking by gel electrophoresis revealed a level of cross-linking of around 45% an d 50% at the different conditions used. In vivo behavior of these modi fied rat erythrocytes revealed that they do not circulate, showing a p redominant localization in the liver. This effect is evident from the concentration (5 mM BS3 or DTSSP) used. Based on these results, BS3 an d DTSSP can be considered as useful tools to cross-link rat erythrocyt e band 3 and to target rat erythrocytes to the liver.