Systematic investigation of orthogonal SPE sample preparation for the LC-MS/MS bioanalysis of a monoclonal antibody after pellet digestion.
BACKGROUND
Increasing assay sensitivity is critical for promoting the application of LC-MS/MS quantitative bioanalysis of therapeutic proteins. A sample processing method that can selectively remove the abundant background peptides in the serum tryptic digests and retain the target peptides can greatly improve the assay sensitivity.


RESULTS
Mixed-mode strong-cation exchange SPE was systematically investigated as an orthogonal sample separation technique to reversed-phase UHPLC for the analysis of a test monoclonal antibody, BMS-986012, in monkey serum after pellet digestion. Strong cation exchange SPE efficiently removed most of the background peptides and reduced the matrix effect and background level in the monitored mass transition channels. As a result, improved sensitivity was observed for the surrogate peptides VVSV and SLIY.


CONCLUSIONS
This orthogonal approach provides a simple and easy-to-develop sample preparation method that can selectively remove most background peptides and extract the target peptides, therefore, improving the LC-MS/MS assay sensitivity.
Bioanalytical Sciences, Analytical & Bioanalytical Development, Bristol-Myers Squibb, Princeton, NJ 08543, USA.
Long
L
Yuan
Anne-Françoise
AF
Aubry
Mark E
ME
Arnold
Qin C
QC
Ji
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000097:blood* / Q000302:isolation & purification / Q000378:metabolism; D001798:Blood Proteins / Q000378:metabolism; D002851:Chromatography, High Pressure Liquid; D000882:Haplorhini; D006801:Humans; D007553:Isotope Labeling; D010455:Peptides / Q000032:analysis / Q000737:chemistry / Q000302:isolation & purification; D052616:Solid Phase Extraction; D053719:Tandem Mass Spectrometry; D014357:Trypsin / Q000378:metabolism
D000911:Antibodies, Monoclonal; D001798:Blood Proteins; D010455:Peptides; D014357:Trypsin