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Separation of Carboxy- and Methemoglobin by Cation Exchange Dextran Gels

1963; Taylor & Francis; Volume: 15; Issue: 4 Linguagem: Inglês

10.3109/00365516309079753

1502-7686

A. M. M. Zade‐Oppen,

Hemoglobin structure and function

Human adult carboxyhemoglobin (Hb,-Co), methemoglobin (MHb,), and mixtures of the pure derivatives were transferred into phosphate buffer pH 6.0 by gel filtration on Sephadex G 25 and then adsorbed on two different ion exchangers with a high degree of swelling. In one case the charged groups were carboxyl, CM-Sephadex C-50, and in the other sulphonate, SE-Sephadex C-50. They were then eluted from the column with a continuously increasing concentration of NaCl in the same buffer.On CM-Sephadex, HbA CO separated into two fractions, probably HbA0, CO and HbA1CO, but this fractionation was poor on SE-Sephadex.Mixtures of Hb, CO and hfHb, gave a good separation on SE-Sephadex, less good on CM-Sephadex. A two-step elution procedure gave a good separation on SE-Sephadex with purities of over 90 per cent in parts of both fractions. The elution behavior indicated that the same result can be obtained by elution with a constant eluant concentration.