Hemosol Inc.

Connaught Laboratories Limited

Anthony A Magnin

Diana H Pliura

Salman Ashraf

Diane E Wiffen

Paul Dennis

Po-Shing Wah

A preselected hemoglobin species is separated from contaminants having a different acidity from that of the preselected hemoglobin species, by an overload displacement chromatography process. To remove more acidic contaminants, the process is conducted under anion exchange conditions. To remove more basic contaminants, the process is conducted under cation exchange conditions. In either case, the exchange column is overloaded to displace the hemoglobin species therefrom with contaminants having greater affinity for the column, and using the impure hemoglobin solution as the displacer.

Purification of hemoglobin by displacement chromatography

Hemoglobin is purified from a crude solution thereof, to obtain an aqueous solution containing at least 99% of a preselected hemoglobin species, by a two stage displacement chromatography process. One of the stages is conducted under anionic exchange conditions, and the other under cationic exchange conditions. In both stages, the exchange column is overloaded to displace the hemoglobin species therefrom with contaminants having greater affinity for the column, and using the impure hemoglobin solution as the displacer. Normally, anionic exchange is conducted first, with contaminants more acidic than the hemoglobin displacing the hemoglobin from the column and themselves remaining attached to the column for separation. The cationic exchange process is conducted second, on the eluent from the first column, and in this stage, the more basic contaminants displace the hemoglobin from the column under overload conditions, to yield a substantially pure hemoglobin solution.

Displacement chromatography process

Pasteurization of immunoglobulin solutions to inactivate viruses without significantly altering the IgG molecules or their physiological activities is obtained with minimal aggregation in the absence of any stabilizer by heating this solution to mildly-elevated temperatures at low ionic strength and mildly acid pH values.

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Willowdale, Canada

Anthony A Magnin