IMAC30 Chips
Add 80 µL of the IMAC30 chip-binding buffer into each well in the bioprocessor set up containing the IMAC30 chips.
Add 20 µL of each ion exchange bead eluate (serum fractions 1–6) to the well for sample binding.
Mix well by shaking for 30 min at RT.
Remove the serum fractions.
Wash the chip spot in the well by adding 150 µL of IMAC30 chip-binding buffer onto each well and shake for 5 min at RT.
Remove buffer.
Repeat washing (steps 5 and 6) once more.
Rinse with water two times by adding 200 µL of Milli-Q water to each well and discard immediately.
Remove the chips from bioprocessor and air-dry the chips for 5 min.
Add 1 µL of 50% sinapinic acid (which is an energy absorbing molecule solution) to each chip spot.
Air-dry the chips for about 10 min.
Add 1 µL of 50% sinapinic acid and air-dry again.
The chip is ready to be read in the PBS IIc mass spectrometer reader.
Remember to process at least one reference control serum concurrently with the patients’ samples on each chip for quality control of chip-to-chip variability.