Separation mode Carrier ampholyte mixture Separation conditions Mobili-zation Coating Interface Sheath liquid MS instrument Analyte Ref.
CIEF
Ampholytes: pH 3–10 (1.5% v/v) in 30:70 glycerol/water (v/v)
Anolyte: 50mM formic acid/1 mM glutamic acid (pH 2.35); catholyte: 100mM NH3/1 mM lysine (pH 10.6); normal polarity (30 kV)
50 mbar
Bare fused silica
Coaxial SL
MeOH/water/acetic acid (50:50:1 v/v/v)
Single quadrupole (Series 1100 MSD)
Ribonuclease A, a-chymotrypsinogen A type II, myoglobin, carbonic anhydrase II, b-lactoglobulin A, trypsin inhibitor
CIEF
Ampholytes: pH 3–10 (1.5% v/v) and pH 9–11 (1% v/v) in 40/60 (v/v) glycerol/water
Anolyte: 50 mM formic acid/1 mM glutamic acid (pH 1.9); catholyte: 100 mM NH3/1 mM arginine (pH 11.3); normal polarity (30 kV);
50 mbar, 30 kV
LPA
Coaxial SL
10 mM AmAc (pH 5.0)
Ion trap (Esquire 3000+)
Recombinant wild-type IFN-γ, single and deletion mutants
CIEF
For SA/ CA Ampholytes: pH 3−10 and pH 5−8 (1.5% v/v in total) in 10 mM AmAc and 15% glycerol (pH 6.7); For ADC Ampholytes: 0.75% (v/v) pH 3−10 and 8−10.5 with a ratio of 1:2
Anolyte: 0.1% (v/v) formic acid (FA), 15% glycerol (pH 3.0); catholyte: 10 mM AmBicarb, 15% glycerol, (pH 10.0); normal polarity (30 kV);
10 mbar, 20 kV
LPA
Nano SL
10 mM AmAc, 10% (v/v) ACN (pH 5.0)
Q-TOF (6545XT)
Recombinant streptavidin (SA), carbonic anhydrase II (CA), interchain cysteine-linked antibody−drug conjugate (ADC)