TY - JOUR
T1 - Purification of human hemoglobin valence intermediates by preparative immobilized pH gradients
AU - Gelfi, Cecilia
AU - Giorgio Righetti, Pier
AU - Rovida, Ermanna
AU - Samaja, Michele
PY - 1987
Y1 - 1987
N2 - We compare three separation techniques for preparative purposes, i.e. ion-exchange chromatography on CM-cellulose, conventional isoelectric focusing in polyacrylamide gel slabs and immobilized pH gradients. The biological system used to test the three methods is a solution containing four hemoglobin (Hb) valence intermediates, i.e. metHb, oxyHb, (α+βO2
)2 and (αO2
β+)2. The ΔpI between the two valence intermediates is 0.04 pH units. Immobilized pH gradients give the best performance in terms of resolving power, total amount of protein which can be loaded and retention of biological activity by the protein (the latter assessed by determination of CO dissociation rates).
AB - We compare three separation techniques for preparative purposes, i.e. ion-exchange chromatography on CM-cellulose, conventional isoelectric focusing in polyacrylamide gel slabs and immobilized pH gradients. The biological system used to test the three methods is a solution containing four hemoglobin (Hb) valence intermediates, i.e. metHb, oxyHb, (α+βO2
)2 and (αO2
β+)2. The ΔpI between the two valence intermediates is 0.04 pH units. Immobilized pH gradients give the best performance in terms of resolving power, total amount of protein which can be loaded and retention of biological activity by the protein (the latter assessed by determination of CO dissociation rates).
KW - Hemoglobin valence intermediates
KW - Immobilized pH gradients
KW - Isoelectric focusing
KW - Preparative electrophoresis
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U2 - 10.1016/0165-022X(87)90016-9
DO - 10.1016/0165-022X(87)90016-9
M3 - Article
C2 - 3680854
AN - SCOPUS:0023195224
VL - 14
SP - 139
EP - 147
JO - Journal of Biochemical and Biophysical Methods
JF - Journal of Biochemical and Biophysical Methods
SN - 0165-022X
IS - 3
ER -