Preparation of low‐affinity red cells with dimethylsulfoxide‐mediated inositol hexaphosphate incorporation: Hemoglobin and ATP recovery using a continuous‐flow method

Incorporation of IHP into red cells decreases oxygen affinity as a result of the binding of this compound to the 2,3‐DPG site of hemoglobin. This investigation describes a continuous‐flow method which utilizes the osmotic pulse technique to transport IHP into RBC. Using this procedure, it is possible to obtain a significant increase in P 50 while maintaining in vitro cellular integrity. For example. IHP incorporation sufficient to cause an increase in the P 50 of 20 mm Hg may be achieved with recovery of approximately 75% of the hemoglobin and with maintenance of ATP levels compatible with good viability. The continuous‐flow method allows uniform treatment of large, unit‐size volumes of red cells with a relatively small quantity of reagents. The final cell product is macrocytic/hypochromic with an increased number of stomatocytes.