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Updated: Jun 19, 2026

Ex Vivo Red Blood Cell Hemolysis Assay for the Evaluation of pH-responsive Endosomolytic Agents for Cytosolic Delivery of Biomacromolecular Drugs
Published on: March 9, 2013
THE EQUILIBRIUM BETWEEN HEMOLYTIC SENSITIZER AND RED BLOOD CELLS IN RELATION TO THE HYDROGEN ION CONCENTRATION
1Hoagland Laboratory, Brooklyn.
Abstract:
1. In a salt-free medium the proportion of the total amount of hemolytic sensitizer present, combined with the homologous cells, reaches a maximum of almost 100 per cent at pH 5.3. On the alkaline side of this point the proportion combined diminishes with the alkalinity and reaches a minimum of approximately 5 per cent at pH 10. On the acid side of pH 5.3 the proportion combined diminishes with the acidity but somewhat less rapidly than for a corresponding increase in alkalinity. 2. The presence of NaCl greatly increases the proportion of sensitizer combined with cells at all reactions except those in the neighborhood of pH 5.3. At this point the combination of sensitizer with cells is independent of the presence of electrolyte. 3. The curves representing the proportion of sensitizer combined or free run almost exactly parallel, both when the sensitizer combines de novo and when it dissociates from combination; therefore, in constant volume, at a given hydrogen ion concentration, and at a given temperature, an equilibrium exists between the amount of sensitizer free and that combined with cells. 4. The combination of sensitizer and cells is related fundamentally to the isoelectric point of the sensitizer. 5. The dissociated ions of the sensitizer, formed either by its acid or its basic dissociation, do not unite with cells. Combination takes place only between the cells and the undissociated molecules of the sensitizer.
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