Metallomics Studies of Human Blood Serum from Treated Bipolar Disorder Patients

In the present work, metallomics studies using biomolecular (matrix-assisted laser desorption ionization time-of-flight tandem mass spectrometry, MALDI-TOF MS/MS) and elemental mass spectrometry (laser ablation inductively coupled plasma mass spectrometry, LA-ICPMS) of human blood serum samples from bipolar disorder (BD) patients compared to controls were performed. The serum samples from three different groups: control (n = 25), BD patients treated with Li (n = 15), and BD patients not treated with Li (n = 10), were pooled according to their groups and separated by two-dimensional polyacrylamide gel electrophoresis (2-D PAGE). Then, in order to determine the metals bound to the protein spots and search for differences among the studied groups, the 2-D gels were analyzed by LA-ICPMS in three distinct modes: bioimaging of metals in gel sections, line scan through the protein spots, and microlocal analysis of selected protein spots. MALDI-TOF MS/MS characterized 32 serum proteins, and they were associated with the metals previously detected. When comparing control and treated BD patient groups, a differentiation in terms of metals bound to proteins was possible to observe. The main metals bound to proteins found in all groups were Na, Mg, Zn, Ca, and Fe. Mn was only detected in the control group; Co was only observed in the control and BD patients treated with Li group. K and Ti were only found in the BD patient groups, and P was only observed in control and BD patients not treated with Li drugs. This exploratory work shows that the association of LA-ICPMS with MALDI-TOF MS/MS is a powerful strategy in metallomics studies applied to determine differences in metal-containing proteins, being able to play an important role on the discovery of potential markers for BD and its treatment with Li in serum samples.