Skip Navigation
Skip to contents

PHRP : Osong Public Health and Research Perspectives



Page Path
HOME > Search
1 "freeze–thaw cycles"
Article category
Publication year
Original Article
Effect of Repeated Freezing and Thawing on Biomarker Stability in Plasma and Serum Samples
Jae-Eun Lee, Shine Young Kim, So-Youn Shin
Osong Public Health Res Perspect. 2015;6(6):357-362.   Published online December 31, 2015
  • 3,292 View
  • 24 Download
  • 45 Crossref
AbstractAbstract PDF
The stability of circulating proteins can be affected by repeated freezing and thawing. The aim of our study was to identify the effect of repeated freezing and thawing on the plasma and serum concentrations of eight proteins [interferon-γ, interleukin (IL)-8, IL-15, IL-17A, matrix metalloproteinase (MMP)-7, tumor necrosis factor-α, vascular endothelial growth factor (VEGF), and VEGF receptor 2 (VEGF-R2)].
We assessed the concentration changes of these proteins in 30 plasma and serum samples subjected to three, four, or five freeze–thaw cycles, and compared these with the concentration changes in the samples that were subjected to two freeze–thaw cycles before analysis.
Repeated freezing and thawing by up to five cycles did not modify the plasma and serum concentrations of interferon-γ, IL-8, and VEGF-R2, while levels of MMP-7, tumor necrosis factor-α, and VEGF were significantly changed in both plasma and serum samples. Moreover, MMP-7 and VEGF concentrations tended to increase with freeze–thaw cycles. They were more elevated in plasma samples (up to about 15%) than in serum samples (up to about 7%), suggesting that serum is the preferred sample type for the analysis of circulating proteins.
This is the first report on the effect of repeated freezing and thawing on plasma concentrations of MMP-7 and VEGF-R2. Our findings propose that researchers should consider the number of freeze–thaw cycles to select plasma or serum samples, depending on the type of analyte.


Citations to this article as recorded by  
  • Reliability, stability during long-term storage, and intra-individual variation of circulating levels of osteopontin, osteoprotegerin, vascular endothelial growth factor-A, and interleukin-17A
    Tomoki Nakamizo, John Cologne, Takeshi Kishi, Tetsuya Takahashi, Mayumi Inoue, Hiroyuki Ryukaku, Tomonori Hayashi, Yoichiro Kusunoki, Saeko Fujiwara, Waka Ohishi
    European Journal of Medical Research.2024;[Epub]     CrossRef
  • Quantitative and qualitative analysis of stability for 16 serum immunoregulators over 50 freeze–thaw cycles
    R. N. Chowdhury, A. Armato, E. Culver, L. Shteynman, C. Kurien, B. Cradin, F. Margolin, T. Nguyen, C. Cardona, N. Kabir, R. M. Garruto, J. K. Lum, K. Wander
    American Journal of Human Biology.2024;[Epub]     CrossRef
  • Dynamics of serum levels of TNF-α in a longitudinal follow-up study in 98 patients with juvenile idiopathic arthritis treated with anti-TNF-α biological drugs
    N. Morozova, M. Zajc Avramovič, G. Markelj, N. Toplak, T. Avčin
    Clinical Rheumatology.2024;[Epub]     CrossRef
  • Sample processing time but not storage time affects complement activation markers C4a, C4d, C3a, iC3b, Bb, C5a, and sC5b-9 levels in EDTA-plasma of individuals at clinical high-risk for psychosis
    Eleftheria Kodosaki, Colm Healy, Jonah F. Byrne, Melanie Föcking, Mary Cannon, Diana O. Perkins, David Cotter, Meike Heurich
    Biomarkers in Neuropsychiatry.2024; 10: 100097.     CrossRef
  • Biobanking for Neurodegenerative Diseases: Challenge for Translational Research and Data Privacy
    Emilia Giannella, Valentino Notarangelo, Caterina Motta, Giulia Sancesario
    The Neuroscientist.2023; 29(2): 190.     CrossRef
  • Keep cool! Observed temperature variations at different process stages of the biobanking workflow – examples from the Leipzig medical biobank
    Juliane Weikert, Angelina Mehrländer, Ronny Baber
    Journal of Laboratory Medicine.2023; 47(2): 69.     CrossRef
  • The Stability of the Anti-Müllerian Hormone in Serum and Plasma Samples under Various Preanalytical Conditions
    Radana Vrzáková, Václav Šimánek, Ondřej Topolčan, Vladimír Vurm, David Slouka, Radek Kučera
    Diagnostics.2023; 13(8): 1501.     CrossRef
  • Differences in the inflammatory proteome of East African and Western European adults and associations with environmental and dietary factors
    Godfrey S Temba, Nadira Vadaq, Vesla Kullaya, Tal Pecht, Paolo Lionetti, Duccio Cavalieri, Joachim L Schultze, Reginald Kavishe, Leo AB Joosten, Andre J van der Ven, Blandina T Mmbaga, Mihai G Netea, Quirijn de Mast
    eLife.2023;[Epub]     CrossRef
  • Influence of Freezing Temperature, Freezing Duration, and Repeated Freeze/Thaw Cycles on Electrophoretic Profiles in the White Stork (Ciconia ciconia)
    Milan Thorel, Yannick Roman, Antoine Leclerc
    Journal of Avian Medicine and Surgery.2023;[Epub]     CrossRef
  • Clinical Spectroscopy: Lost in Translation?
    James M. Cameron, Christopher Rinaldi, Samantha H. Rutherford, Alexandra Sala, Ashton G. Theakstone, Matthew J. Baker
    Applied Spectroscopy.2022; 76(4): 393.     CrossRef
  • Correlation of Repeat Measurements of 27 Candidate Protein Markers for Colorectal Cancer Screening Taken Three Years and Multiple Freeze–Thaw Cycles Apart
    Megha Bhardwaj, Petra Schrotz-King, Hermann Brenner
    Life.2022; 12(3): 359.     CrossRef
  • Development of quinone linked immunosorbent assay (QuLISA) based on using Folin’s reagent as a non-enzymatic tag: Application to analysis of food allergens
    Mahmoud El-Maghrabey, Yudai Sato, Fatema Kaladari, Naoya Kishikawa, Naotaka Kuroda
    Sensors and Actuators B: Chemical.2022; 368: 132167.     CrossRef
  • A systematic review and meta-analysis of preanalytical factors and methodological differences influencing the measurement of circulating vascular endothelial growth factor
    Ulrika Sjöbom, Anders K. Nilsson, Hanna Gyllensten, Ann Hellström, Chatarina Löfqvist, Angela J. Glading
    PLOS ONE.2022; 17(7): e0270232.     CrossRef
  • Cytokines: From Clinical Significance to Quantification
    Chao Liu, Dewei Chu, Kourosh Kalantar‐Zadeh, Jacob George, Howard A. Young, Guozhen Liu
    Advanced Science.2021;[Epub]     CrossRef
  • Microheterogeneity and preanalytical stability of protein biomarkers of inflammation and renal function
    Jie Gao, Arve Ulvik, Adrian McCann, Per Magne Ueland, Klaus Meyer
    Talanta.2021; 223: 121774.     CrossRef
  • The effect of pre-analytical handling on the stability of fractalkine, monocyte chemoattractant protein 1 (MCP1), interleukin 6 and interleukin 8 in samples of human cerebrospinal fluid
    Winnie Charlotte Pedersen Mortensen, Laila Bendix, Hanne Irene Jensen, Claus Varnum, Lasse Enkebølle Rasmussen, Jørgen T. Lauridsen, Nis Borbye-Lorenzen, Kristin Skogstrand, Palle Toft, Henrik Bjarke Vaegter, Morten Rune Blichfeldt-Eckhardt
    Journal of Immunological Methods.2021; 494: 113057.     CrossRef
  • Standardizing salivary lactoferrin measurements to obtain a robust diagnostic biomarker for Alzheimer's disease
    Fernando Bartolome, Gorka Orive, Eva Carro
    Alzheimer's & Dementia: Diagnosis, Assessment & Di.2021;[Epub]     CrossRef
  • Pre-analytical considerations in biomarker research: focus on cardiovascular disease
    Elena Revuelta-López, Jaume Barallat, Adriana Cserkóová, Carolina Gálvez-Montón, Allan S. Jaffe, James L. Januzzi, Antoni Bayes-Genis
    Clinical Chemistry and Laboratory Medicine (CCLM).2021; 59(11): 1747.     CrossRef
  • Vaginal cytokine profile and microbiota before and after lubricant use compared with condomless vaginal sex: a preliminary observational study
    Susan Tuddenham, Christina A. Stennett, Richard A. Cone, Jacques Ravel, Andrew N. Macintyre, Khalil G. Ghanem, Xin He, Rebecca M. Brotman
    BMC Infectious Diseases.2021;[Epub]     CrossRef
  • Proximity Extension Assay in Combination with Next-Generation Sequencing for High-throughput Proteome-wide Analysis
    Lotta Wik, Niklas Nordberg, John Broberg, Johan Björkesten, Erika Assarsson, Sara Henriksson, Ida Grundberg, Erik Pettersson, Christina Westerberg, Elin Liljeroth, Adam Falck, Martin Lundberg
    Molecular & Cellular Proteomics.2021; 20: 100168.     CrossRef
  • Serum or plasma? An old question looking for new answers
    Mario Plebani, Giuseppe Banfi, Sergio Bernardini, Francesco Bondanini, Laura Conti, Romolo Dorizzi, Fulvio Enrico Ferrara, Rita Mancini, Tommaso Trenti
    Clinical Chemistry and Laboratory Medicine (CCLM).2020; 58(2): 178.     CrossRef
  • Association Between Intravenous Fluid Bolus and Biomarker Trajectory During Prehospital Care
    Emily B. Brant, Jason Kennedy, Christian Martin-Gill, Vanessa Jackson, Octavia M. Peck Palmer, Clifton W. Callaway, Jeremy M. Kahn, Donald M. Yealy, Derek C. Angus, Christopher W. Seymour
    Prehospital Emergency Care.2020; 24(2): 196.     CrossRef
  • Evaluating sample stability in the clinical laboratory with the help of linear and non-linear regression analysis
    Joachim K.W. Pum
    Clinical Chemistry and Laboratory Medicine (CCLM).2020; 58(2): 188.     CrossRef
  • Optimization of Saliva Collection and Immunochromatographic Detection of Salivary Pepsin for Point-of-Care Testing of Laryngopharyngeal Reflux
    Young Ju Lee, Jiyoon Kwon, Sanggyeong Shin, Young-Gyu Eun, Jae Ho Shin, Gi-Ja Lee
    Sensors.2020; 20(1): 325.     CrossRef
  • Stability of 12 T-helper cell-associated cytokines in human serum under different pre-analytical conditions
    Ulrik Stervbo, Sharon Bajda, Patrizia Wehler, Benjamin J. Rohn, Melanie Streichhahn, Sehriban Temizsoy, Eva Kohut, Toralf Roch, Richard Viebahn, Timm H. Westhoff, Nina Babel
    Cytokine.2020; 129: 155044.     CrossRef
  • Quinone-based antibody labeling reagent for enzyme-free chemiluminescent immunoassays. Application to avidin and biotinylated anti-rabbit IgG labeling
    Mahmoud El-Maghrabey, Naoya Kishikawa, Shiori Harada, Kaname Ohyama, Naotaka Kuroda
    Biosensors and Bioelectronics.2020; 160: 112215.     CrossRef
  • Standardized approaches for clinical sampling and endpoint ascertainment in tuberculous meningitis studies
    Ursula K Rohlwink, Felicia C Chow, Sean Wasserman, Sofiati Dian, Rachel PJ Lai, Lidya Chaidir, Raph L Hamers, Robert J Wilkinson, David R Boulware, Fiona V Cresswell, Arjan van Laarhoven
    Wellcome Open Research.2020; 4: 204.     CrossRef
  • Exploring pre-analytical factors for the optimisation of serum diagnostics: Progressing the clinical utility of ATR-FTIR spectroscopy
    James M. Cameron, Holly J. Butler, David J. Anderson, Loren Christie, Lily Confield, Katie E. Spalding, Duncan Finlayson, Stuart Murray, Zanib Panni, Christopher Rinaldi, Alexandra Sala, Ashton G. Theakstone, Matthew J. Baker
    Vibrational Spectroscopy.2020; 109: 103092.     CrossRef
  • QuantiFERON-TB Gold Plus with Chemiluminescence Immunoassay: Do We Need a Higher Cutoff?
    Francesco Bisognin, Giulia Lombardi, Maria Carla Re, Paola Dal Monte, Geoffrey A. Land
    Journal of Clinical Microbiology.2020;[Epub]     CrossRef
  • Stability of serum biochemical markers during standard long-term storage and with a single thawing
    V. A. Kozlova, V. A. Metelskaya, M. S. Pokrovskaya, I. A. Efimova, O. A. Litinskaya, V. A. Kutsenko, E. B. Yarovaya, S. A. Shalnova, O. M. Drapkina
    Cardiovascular Therapy and Prevention.2020; 19(6): 2736.     CrossRef
  • Development and application of a porcine specific ELISA for the quantification of soluble CD163
    J. Alex Pasternak, Daniel J. MacPhee, John C.S. Harding
    Veterinary Immunology and Immunopathology.2019; 210: 60.     CrossRef
  • Dynamics of circulating TNF during adalimumab treatment using a drug-tolerant TNF assay
    Lea C. Berkhout, Merel J. l’Ami, Jill Ruwaard, Margreet H. Hart, Pleuni Ooijevaar-de Heer, Karien Bloem, Michael T. Nurmohamed, Ronald F. van Vollenhoven, Maarten Boers, Daniel F. Alvarez, Catherine H. Smith, Gerrit J. Wolbink, Theo Rispens
    Science Translational Medicine.2019;[Epub]     CrossRef
  • Standardized approaches for clinical sampling and endpoint ascertainment in tuberculous meningitis studies
    Ursula K Rohlwink, Felicia C Chow, Sean Wasserman, Sofiati Dian, Rachel PJ Lai, Lidya Chaidir, Raph L Hamers, Robert J Wilkinson, David R Boulware, Fiona V Cresswell, Arjan van Laarhoven
    Wellcome Open Research.2019; 4: 204.     CrossRef
  • The Effect of Anticoagulants, Temperature, and Time on the Human Plasma Metabolome and Lipidome from Healthy Donors as Determined by Liquid Chromatography-Mass Spectrometry
    Manoj Khadka, Andrei Todor, Kristal M. Maner-Smith, Jennifer K. Colucci, ViLinh Tran, David A. Gaul, Evan J. Anderson, Muktha S. Natrajan, Nadine Rouphael, Mark J. Mulligan, Circe E. McDonald, Mehul Suthar, Shuzhao Li, Eric A. Ortlund
    Biomolecules.2019; 9(5): 200.     CrossRef
  • A caspase-6-cleaved fragment of Glial Fibrillary Acidic Protein as a potential serological biomarker of CNS injury after cardiac arrest
    Ditte S. Jonesco, Christian Hassager, Martin Frydland, Jesper Kjærgaard, Morten Karsdal, Kim Henriksen, Stephen D. Ginsberg
    PLOS ONE.2019; 14(11): e0224633.     CrossRef
  • Angiogenic Factor and Cytokine Analysis among Patients Treated with Adjuvant VEGFR TKIs in Resected Renal Cell Carcinoma
    Wenxin Xu, Maneka Puligandla, Judith Manola, Andrea J. Bullock, Daniel Tamasauskas, David F. McDermott, Michael B. Atkins, Naomi B. Haas, Keith Flaherty, Robert G. Uzzo, Janice P. Dutcher, Robert S. DiPaola, Rupal S. Bhatt
    Clinical Cancer Research.2019; 25(20): 6098.     CrossRef
  • Stability of important antibodies for kidney disease: pre-analytic methodological considerations
    Qiuxia Han, Songyan Li, Bo Fu, Dongwei Liu, Maoqing Wu, Xiaoli Yang, Guangyan Cai, Zhangsuo Liu, Xiangmei Chen, Hanyu Zhu
    PeerJ.2018; 6: e5178.     CrossRef
  • The association of angiogenic factors and chronic kidney disease
    Christopher E. Anderson, L. Lee Hamm, Gem Batuman, Damodar R. Kumbala, Chung-Shiuan Chen, Swapna G. Kallu, Ravi Siriki, Shilpa Gadde, Myra A. Kleinpeter, N. Kevin Krane, Eric E. Simon, Jiang He, Jing Chen
    BMC Nephrology.2018;[Epub]     CrossRef
  • Strong impact on plasma protein profiles by precentrifugation delay but not by repeated freeze-thaw cycles, as analyzed using multiplex proximity extension assays
    Qiujin Shen, Johan Björkesten, Joakim Galli, Daniel Ekman, John Broberg, Niklas Nordberg, Annika Tillander, Masood Kamali-Moghaddam, Gunnel Tybring, Ulf Landegren
    Clinical Chemistry and Laboratory Medicine (CCLM).2018; 56(4): 582.     CrossRef
  • Plasma-Derived Inflammatory Proteins Predict Oral Squamous Cell Carcinoma
    Kelly Yi Ping Liu, Xian Jun David Lu, Yuqi Sarah Zhu, Nhu Le, Hugh Kim, Catherine F. Poh
    Frontiers in Oncology.2018;[Epub]     CrossRef
  • Is vascular endothelial growth factor a useful biomarker in giant cell arteritis?
    Nicola Goodfellow, Julien Morlet, Surjeet Singh, Afsie Sabokbar, Andrew Hutchings, Vanshika Sharma, Jana Vaskova, Shauna Masters, Allahdad Zarei, Raashid Luqmani
    RMD Open.2017; 3(1): e000353.     CrossRef
  • Indices of immune function used by ecologists are mostly unaffected by repeated freeze-thaw cycles and methodological deviations
    Arne Hegemann, Sara Pardal, Kevin D. Matson
    Frontiers in Zoology.2017;[Epub]     CrossRef
  • The Enhanced Liver Fibrosis (ELF) Panel: Analyte Stability Under Common Sample Storage Conditions Used in Clinical Practice
    Oliver J Kennedy, Julie Parkes, Sudeep Tanwar, Paul M Trembling, William M Rosenberg
    The Journal of Applied Laboratory Medicine.2017; 1(6): 720.     CrossRef
  • Instability of Plasma and Serum Progastrin-Releasing Peptide During Repeated Freezing and Thawing
    Jae-Eun Lee, Jin-Hyun Lee, Maria Hong, Seul-Ki Park, Ji-In Yu, So-Youn Shin, Shine Young Kim
    Osong Public Health and Research Perspectives.2016; 7(6): 351.     CrossRef
  • Effects of Long-Term Storage Time and Original Sampling Month on Biobank Plasma Protein Concentrations
    Stefan Enroth, Göran Hallmans, Kjell Grankvist, Ulf Gyllensten
    EBioMedicine.2016; 12: 309.     CrossRef

PHRP : Osong Public Health and Research Perspectives