Isolation and Characterization of Erythrocyte and Parasite Membranes from Rhesus Red Cells Infected with P. knowlesi.
Abstract
Extension of the fractionation approach given above will allow us to identify and characterize proteins typically associated with the parasite surface membrane and other subcellular organeles. Furthermore, we shall be able to eliminate and isolate erythrocyte membrane contaminants or vesicles originating from the vacuolar membrane by employing affinity density perturbation. In this technique immunoglobulin against normal erythrocyte membrane proteins (see section 2.1.1.3.) will be coupled to latex beads which will then be reacted with the parasite homogenate. This will allow us to selectively increase the density of membrane vesicles containing proteins characteristics of normal erythrocyte membranes and to isolate those membranes by centrifugation in a density gradient. Our current immunization scheme using purified intracecellular plasmodium knowlesi parasites in CFA allows us to produce a high titered anti-serum in the natural host of this parasite. The serum strongly agglutinates isolated parasites and infected erythrocytes (SICA) and is also suitable for immunochemical analyses. Comparative analyses of membrane proteins from normal and parasitized erythrocytes and purified parasite indicate that there is only minimal cross-contamination when infected cells are subfractionated.
Document Details
- Document Type
- Technical Report
- Publication Date
- Jun 01, 1981
- Accession Number
- ADA140277
Entities
People
- D. F. H. Wallach
Organizations
- Tufts Medical Center