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Serum autoantibody measurement aids in diagnosing and monitoring various autoimmune conditions. Defining autoantibody stability limits can improve laboratory process quality. Here, we define short-term stability in a refrigerator, long-term stability in a freezer, and the effect of freeze-thaw cycles to improve autoantibody testing procedures. Seventy-nine residual serum samples were used to assess the stability of 11 autoantibodies (anti-dsDNA, anti-Ro52, anti-Ro60, anti-SSB, anti-RNP, anti-Sm, anti-aCL-IgG, anti-tTG-IgA, anti-tTG-IgG,