In recent years the blood proteome has been increasingly researched for biomarkers for early detection of colorectal cancer (CRC). Blood samples from screening studies are often subject to preanalytical variability and repeated freeze−thaw cycles. We aimed to assess the correlation of repeat measurements of 27 candidate protein markers for CRC screening taken three years and multiple freeze−thaw cycles apart. The concentrations of 27 protein markers were measured in plasma samples of 39 newly detected CRC cases from a cohort of 9245 participants of screening colonoscopies. The proteins were measured using proximity extension assays (PEA) carried out on the same set of samples twice, three years apart, with an average of three freeze−thaw cycles in between the two measurements. Pearson’s product moment correlation coefficients were calculated. Correlation coefficients ranged from +0.43 to +0.97, with a median of 0.67 and an interquartile range of +0.58 to +0.84, with all p-values of correlation being <0.01 (<0.0005 for 22 proteins, <0.001 for 4 proteins). Repeat measurements of the 27 protein biomarkers for CRC screening performed three years later, and on average three freeze−thaw cycles apart, showed moderate to high levels of correlation. Apart from the effects of freeze−thaw cycles, slightly different preprocessing performed on the data may have contributed to recorded differences between measurements.
Keywords: colorectal cancer; freeze–thaw cycles; plasma proteins; proximity extension assays; sample handling.