Encapsulation of Alcohol Dehydrogenase and Acetaldehyde Dehydrogenase into Human Erythrocytes by an Electroporation Procedure
The optimal conditions for electroporation/resealing loading of alcohol dehydrogenase (ADH) and acetaldehyde dehydrogenase (ALDH) into human erythrocytes were first established (37ºC, 300V, 1ms pulse time, 8 pulses every 15min, and 1h resealing). At a fixed enzyme level (100U ADH or 3U ALDH per mL erythrocyte suspension), the encapsulation yield was 26.2% for ADH and 38.3% for ALDH. Carrier cell recoveries were 82–85%. Cell volumes increase while hemoglobin content decrease as a consequence of the electroporation/resealing process (i.e. a decreased cell hemoglobin concentration). Although a lower hypotonic resistance of loaded erythrocytes (all along the osmotic fragility curves) was observed, the non-alteration of the oxygen transport capability (as given by the oxygen equilibrium curves) suggest the electroporation/resealing process as a convenient alternative to the hypotonic-dialysis method for the preparation of ADH- and ALDH-erythrocytes.
KeywordsAlcohol Dehydrogenase Mean Cell Volume Acetaldehyde Dehydrogenase Human RBCs Oxyhemoglobin Dissociation Curve
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