Quantitative determination of noncovalently bound acridinium in protein conjugates by liquid chromatography/electrospray ion trap mass spectrometry.
A sensitive and robust liquid chromatography/electrospray ion trap mass spectrometry (LC/MS/MS) method has been developed for the quantitative determination of noncovalently bound acridinium free acid in protein-acridinium conjugates. The lower level of quantitation (LOQ) for acridinium free acid was determined to be 0.6 ng. The assay was validated with a linear concentration range of 0.6-60 ng. The method requires minimum sample handling and is specific, reproducible, and provides a new aspect for protein-acridinium conjugate characterization.
Adamczyk
M
M
Department of Chemistry (9NM), Abbott Diagnostics Division, Abbott Laboratories, 100 Abbott Park Road, Abbott Park, IL 60064-6016, USA. maciej.adamczyk@abbbott.com
Gebler
J C
JC
Shreder
K
K
Wu
J
J
D000166:Acridines / Q000032:analysis* / Q000302:isolation & purification; D002138:Calibration; D002851:Chromatography, High Pressure Liquid; D011506:Proteins / Q000032:analysis* / Q000302:isolation & purification; D021241:Spectrometry, Mass, Electrospray Ionization; D013388:Succinimides / Q000032:analysis* / Q000302:isolation & purification
D000166:Acridines; C066889:N-hydroxysuccinimide acridinium ester; D011506:Proteins; D013388:Succinimides