Influence of Storage and Inter- and Intra-Assay Variability on the Measurement of Inflammatory Biomarkers in Population-Based Biobanking

Robert P. van Waateringe, Anneke C. Muller Kobold, Jana V. van Vliet-Ostaptchouk, Melanie M. van der Klauw, Jan Koerts, Gabriele Anton, Annette Peters, Gerlinde Trischler, Kirsti Kvaloy, Marit Naess, Vibeke Videm, Kristian Hveem, Melanie Waldenberger, Wolfgang Koenig, Bruce H. R. Wolffenbuttel*

*Corresponding author for this work

Research output: Contribution to journalArticleAcademicpeer-review

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Background: In the present study, we examined the effect of sample storage on the reproducibility of several inflammatory biomarkers, including high-sensitivity C-reactive protein (hsCRP), high-sensitivity interleukin-6 (hsIL6), and high-sensitivity tumor necrosis factor alpha (hsTNF). In addition, we assessed inter- and intra-assay variability between collaborating biobanks. 

Methods: In total, 240 fasting plasma samples were obtained from the LifeLines biobank. Samples had been stored for less than 2 or more than 4 years at -80 degrees C. Measurements were performed at three different laboratories. hsCRP was measured by immunonephelometry and ELISA, hsIL6, and hsTNF samples were measured with ELISAs from two different manufacturers. For confirmation, similar analyses were performed on samples obtained from a subpopulation of 80 obese individuals. Passing-Bablok regression analysis and Bland-Altman plots were used to compare the results. 

Results: We observed good stability of samples stored at -80 degrees C. hsCRP measured on the day of blood draw was similar to levels measured after more than 4 years of storage. There were small interlaboratory differences with the R&D ELISAs for hsIL6 and hsTNF. We found a linear correlation between the Bender Medsystems ELISA and the R&D ELISA for hsIL6, with significantly higher levels measured with the R&D ELISA. Over 90% of hsTNF samples measured with the IBL ELISA were below the detection limit of 0.13ng/L, rendering this assay unsuitable for large-scale analysis. Similar results were found in the confirmation study. 

Conclusion: In summary, plasma hsCRP showed good stability in samples stored for either less than 2 years or more than 4 years at -80 degrees C. Both the R&D and Bender Medsystems for hsIL6 measurement yielded similar results. The IBL hsTNF assay is not suited for use in biobanking samples. Assays for the measurement of inflammatory biomarker assays should be rigorously tested before large sample sets are measured.

Original languageEnglish
Number of pages7
JournalBiopreservation and biobanking
Issue number6
Publication statusPublished - 1-Dec-2017


  • biobanking
  • assay
  • storage
  • variability
  • nephelometry

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