Standard -20 °C freezer storage protocols may cause substantial plasma renin cryoactivation

To assess the appropriate preanalytical process for storage of plasma for renin concentration analysis. This study was initiated due to the wide variation in preanalytical handling of samples observed within our network, particularly with respect to freezing for longer term storage.

Pooled plasma from patient samples was analysed immediately post separation for renin concentration (n=30, concentration 4.0-204 mIU/L). Aliquots from these samples were frozen in a -20 °C freezer and then analysed, with the renin concentration compared to the respective baseline concentration. Comparisons were also made to: aliquots snap frozen using a dry ice/acetone bath, aliquots stored at room temperature, and aliquots stored at 4 °C. Subsequent experiments investigated the potential sources of cryoactivation observed in these initial studies.

Substantial and highly variable cryoactivation was observed in samples frozen using a -20 °C freezer, with renin concentration increasing over 300% from baseline in some samples (median 21.3%). This cryoactivation could be prevented by snap freezing samples. Subsequent experiments determined that long term storage in a -20 °C freezer could prevent cryoactivation provided samples were initially frozen rapidly in a -70 °C freezer. Rapid defrosting of samples was not required to prevent cryoactivation.

Standard -20 °C freezers may not be appropriate for freezing samples for renin analysis. Laboratories should consider snap freezing their samples using a -70 °C freezer or similar to avoid cryoactivation of renin.