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The use of a methyl group is optimum in that it is small enough to not influence the overall activity of the molecule, while the rationale behind the location of the modification is to protect the most common degradation location of the peptides, i.e., the peptide bond

However, in epidemiological studies researchers need to consider the frequency and range of detection of the biomarker, the rarity of the outcome, and the accuracy of the exposure estimate of a given biomarker