Analysis of Metabolomics Response to Tick Saliva in Human Cells.

This chapter describes a workflow for the analysis of the metabolomic response of the human basophilic cell line KU812 exposed to Ixodes ricinus tick saliva using ¹H-nuclear magnetic resonance (NMR) spectroscopy. The protocol includes procedures for cell culture, harvesting, washing, and metabolite extraction, followed by the separation of aqueous and lipid fractions. Detailed instructions for sample preparation and NMR data acquisition are provided for both fractions. Spectral processing and metabolite quantification are performed using targeted profiling approaches based on constrained line-shape fitting, enabling accurate analysis of complex biological samples. Metabolite identification is supported by spectral databases and literature references, and absolute concentrations are obtained using internal standard-based quantification methods. Finally, data normalization based on cell number ensures comparability across samples. Overall, this protocol provides a robust and reproducible strategy for investigating metabolic alterations induced by tick saliva in human cells.