A Sensitive Method for Detecting Sudunit Dissociation of Abnormal Hemoglabin

Dissociation of abnormal hemoglobin (HbA) by gel permeation employing Sephadex G-75 column. Equimolar solutions(5чM)of Hb A and an abnolmal hemoglobin in 0.01 M phosphate buffer, 7.00 containing 0.09 M NaCl and a trace amount of KCM, are introduced into the column (1.0×60　cm,upward flow) by a peristatic pump (flow rate 0.25 ml/min)　one after another. The absorbance at 280 nm or Soret band of the effluent is continuously monitored. The difference in the degree of dissociation is detected by the change in the absorbance at the interface where the two hemoglobins meet. The method developed here has been applied to fifteen different abnormal hemoglobins； twelve of them (80％) have shown either accelerated or reduced dissociation. The method is highly sensitive and specific for the detection of abnormal hemoglobins.

Abnormal hemoglobin

Subunit dissociation