Study of the vesicles released during conservation of red cells

The purpose of this study was to determine the protein composition of the vesicles released from senescent erythrocytes and its variation during their experimental conservation at +4 degrees C in citrate phosphate dextrose (CPD), over a period of 8 weeks. Techniques employed included electron microscopy, SDS-PAGE and immunoblotting. Electron microscopic observation of senescent erythrocytes showed the simultaneous release of one or several vesicles of varying size and shape, while close examination of individual vesicles revealed a slack membrane structure and the presence of band 3 protein. SDS-PAGE and immunoblotting showed the vesicles to be composed mainly of band 3 and its breakdown products and to be lacking in spectrin. Use of specific antibodies demonstrated the presence of free haemoglobin, immunoglobulin G (IgG) and fragment C3b of complement. During conservation for 8 weeks, the concentration of band 3 protein decreased, while the concentrations of IgG and C3b increased and there was no apparent variation in haemoglobin levels.