Stability of solutions of human granulocyte lactate dehydrogenase stored at different temperatures

Granulocyte suspensions from eight healthy individuals were lysed ultrasonically in 0.15 M NaCl or serum. The leukolysates were tested for lactate dehydrogenase after being stored at 22 C, 5 C, -20 C and -196 C for one, two, three, four and seven days. The LDH content of granulocytes was 291 IU/10(10) cells (mean) and the enzyme had a characteristic isoenzyme pattern rich in LDH-5 with stepwise increases in each fraction from LDH-1 to LDH-5. The enzyme was unstable in saline solutions. This was marked at -20 C and related to the duration of storage. All the isoenzyme fractions showed significant inactivation. A mean 10% loss of activity occurred at -196 C. This loss was independent of the duration of storage and related to the process of freezing and thawing. Moderate inactivation occurred at room temperature while refrigerated samples were stable for four days. Serum protected the enzyme from denaturation and samples stored at -20, -196 and 22 C were stable for the seven days of the experiment.