Impact of single freeze-thaw cycles on human serum proteins: Implications for mass spectrometry biomarker validation.

Authors

Thorben Sauer, Martina Oberländer, Regina Maushagen, Helene Radloff, Ruediger Braun, Kim Honselmann, Ulrich Wellner, Jens Habermann, Claudia Benecke, Tobias Keck, Silke Szymczak, Timo Gemoll

Published in

iScience. Volume 29. Issue 6. Pages 115934. Jun 19, 2026. Epub Apr 28, 2026.

Abstract

Validation of biomarkers for clinical diagnostics requires high-quality samples and standardized protocols. This study investigated the effect of a single freeze-and-thaw (FT) cycle on the quality of liquid nitrogen-stored clinical serum samples. Using mass spectrometry, serum protein abundance was analyzed in a human test cohort of 99 samples and a validation cohort 109 samples. After preprocessing, 213 proteins were analyzed in the test cohort and 248 in the validation cohort. Thirty proteins changed significantly in their measurability after freeze-thaw in the test cohort, and 11 were confirmed in the validation cohort. Among them, CP and IGHV6-1 showed potential for distinguishing malignant from benign pancreatic disease. No association was found between protein intensity and storage duration. Overall, the findings show that freeze-thaw cycles are an important pre-analytical factor that should be controlled when translating protein biomarkers into clinical practice.

PMID:
42205694
Bibliographic data and abstract were imported from PubMed on 12 Sep 2026.

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