Encapsulation of concentrated protein into erythrocyte porated by continuous-wave ultrasound.
A procedure of continuous-wave ultrasound (US)-induced hemolysis and reseal in solution containing water soluble protein was applied to a method for encapsulating concentrated protein solutions into resealed rat erythrocyte ghosts. To find a condition yielding a higher mean corpuscular concentration of encapsulated protein (MCC), we investigated the value of MCCs for various conditions. Additions of a small amount of plasma, Ca(2+) and Mg(2+) significantly increased MCC, whereas these additives did not alter the degree of hemolysis. It was suggested that plasma protect the molecular damages by the US, and that Ca(2+) and Mg(2+) physically stabilized the lipids of the erythrocyte membrane to fuse and reseal the pore induced by US. A maximal MCC of approximately 50 mg/mL, which is 2.5 times the reported maximum amount encapsulated by the osmotic dialysis method, was obtained without a blood-washing procedure.
Yamagata
Koutarou
K
Department of Life and Environmental Sciences, Faculty of Engineering, Chiba Institute of Technology, Chiba, Japan.
Kawasaki
Emi
E
Kawarai
Hideki
H
Iino
Masaaki
M
D000818:Animals; D002118:Calcium / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D004337:Drug Carriers; D004910:Erythrocyte Membrane / Q000187:drug effects / Q000648:ultrastructure; D006400:Hematocrit; D008274:Magnesium / Q000494:pharmacology; D008855:Microscopy, Electron, Scanning; D011506:Proteins / Q000008:administration & dosage*; D051381:Rats; D017208:Rats, Wistar; D012710:Serum Albumin, Bovine / Q000008:administration & dosage; D013997:Time Factors; D014465:Ultrasonics
D004337:Drug Carriers; D011506:Proteins; D012710:Serum Albumin, Bovine; D008274:Magnesium; D002118:Calcium