Purification and quantitative determination of carboxymethylchitin incorporation into submicron bilayer-lipid membrane artificial cells (liposomes) encapsulating hemoglobin.
The separation of carboxymethylchitin-coated hemoglobin-loaded liposomes (CMC-LEHb)s from the non-adsorbed CMC has been achieved by gel chromatography. This purification takes place at the physiological pH of 7.4 favoring HbO2 preservation. A comparative study between experimental techniques for the quantitative determination of the absorption of carboxymethylchitin (CMC) onto liposomes encapsulating hemoglobin (LEHb)s has been conducted. Results suggest that FT-IR spectroscopy gives a more accurate quantitative absorption index while the chitinase-based enzymatic assay should be used as a qualitative detection tool. Quantitative bilayer and surface characterization show that the RBC membrane composition has been closely simulated by that of CMC-LEHbs in terms of total lipids and carbohydrates at 87.8% (phospholipids and cholesterol) and 12.2% CMC respectively.
Mobed
M
M
Artificial Cells and Organs Research Centre, McGill University, Montreal, Canada.
Chang
T M
TM
D000818:Animals; D001802:Blood Substitutes / Q000737:chemistry / Q000302:isolation & purification*; D002417:Cattle; D002686:Chitin / Q000031:analogs & derivatives; D002847:Chromatography, Agarose; D006454:Hemoglobins / Q000302:isolation & purification*; D008051:Lipid Bilayers / Q000302:isolation & purification; D008081:Liposomes; D008854:Microscopy, Electron
D001802:Blood Substitutes; D006454:Hemoglobins; D008051:Lipid Bilayers; D008081:Liposomes; D002686:Chitin; C044872:O-(carboxymethyl)chitin