Partitioning behavior of erythrocytes in aqueous two-phase systems containing hydroxypropyl starch and polyethylene glycol.
The partitioning behavior of erythrocytes in Reppal PES 200 (a hydroxypropyl starch produced by Reppe Glykos AB, Växjö, Sweden)-polyethylene glycol (PEG) and in dextran (Dx)-PEG aqueous phase systems made isotonic with phosphate is similar in a number of ways: (i) There is a correlation between the relative electrophoretic mobilities and partition ratios, P, of red blood cells from different species; (ii) The cell P is reduced when, at constant polymer concentrations, phosphate is systematically replaced by sodium chloride (with the total concentration isotonic); (iii) The cell P is increased with reduced polymer concentrations (decreased interfacial tensions); (iv) Treatment of erythrocytes with neuraminidase results in a reduced P value; (v) Rat red cells of different ages can be fractionated by counter-current distribution; and (vi) Differences between red blood cells from genetically distinct rats or between humans can be detected. Aquaphase (a hydroxypropyl starch marketed by Perstorp AB, Lund, Sweden) has been tested as in ii-iv above with analogous results. The partitioning behavior of erythrocytes in PES-PEG and Dx-PEG aqueous phase systems containing sodium chloride differs in a number of ways: (vii) The correlation, apparent in Dx-PEG systems, between the P value of red blood cells from different species and the ratio of their membrane poly- to monounsaturated fatty acids is absent in PES-PEG systems. It is replaced by a correlation as in i; (viii) The increase in P value in Dx-PEG observed from red blood cells after treatment with neuraminidase is replaced by a decrease in P value in PES-PEG or Aquaphase-PEG systems. We conclude that PES (and Aquaphase) can be substitutes for dextran in cell partitioning studies when charge-sensitive phases are used (e.g., those containing phosphate) while separations based on properties reflected by Dx-PEG systems containing sodium chloride are not duplicated by PES-PEG (and probably not by Aquaphase-PEG). The hydroxypropyl starch-PEG systems containing sodium chloride, unlike the analogous Dx-PEG systems, have a significant electrostatic potential difference between the phases.
Walter
H
H
Laboratory of Chemical Biology, Veterans Administration Medical Center, Long Beach, CA 90822.
Krob
E J
EJ
D000375:Aging / Q000097:blood; D000818:Animals; D002860:Chromium Radioisotopes; D003377:Countercurrent Distribution; D004285:Dogs; D004912:Erythrocytes / Q000032:analysis*; D006801:Humans; D066298:In Vitro Techniques; D008297:Male; D009439:Neuraminidase / Q000494:pharmacology; D011092:Polyethylene Glycols; D051381:Rats; D011918:Rats, Inbred SHR; D011919:Rats, Inbred Strains; D011921:Rats, Inbred WKY; D012756:Sheep; D012965:Sodium Chloride / Q000097:blood; D013045:Species Specificity; D013213:Starch; D013552:Swine
D002860:Chromium Radioisotopes; D011092:Polyethylene Glycols; D012965:Sodium Chloride; D013213:Starch; D009439:Neuraminidase