Quantitative aspects of the analysis of the monoclonal antibody trastuzumab using high-performance liquid chromatography coupled with electrospray mass spectrometry.
The analytical possibilities of quantification of the intact monoclonal antibody trastuzumab by high-performance liquid chromatography coupled with electrospray mass spectrometry (HPLC-ESI-MS) were investigated. To clarify the results obtained by LC-MS, complementary experiments were performed using direct UV-spectrophotometry and high-performance liquid chromatography coupled with ultraviolet detection (HPLC-UV). A polystyrene-divinylbenzene (POROS) column was applied with gradient elution using formic acid 0.08% (v/v) in water and formic acid 0.08% (v/v) in acetonitrile as mobile phase for chromatographic analysis. Quantification on LC-MS was performed by using the peak area of the total ion current (TIC) chromatograms of one charge state. Non-linearity and sensitivity loss were the major limitations observed with the LC-MS method, of which the non-linearity is most likely caused by detector saturation. The sensitivity loss during analysis could be reduced by lowering the MS source temperature. This parameter is critical in creating a robust LC-MS system for the quantitative analysis of trastuzumab.
Damen
Carola W N
CW
Department of Pharmacy & Pharmacology, Slotervaart Hospital/The Netherlands Cancer Institute, Louwesweg 6, 1066 EC Amsterdam, The Netherlands. carola.damen@slz.nl
Rosing
Hilde
H
Schellens
Jan H M
JH
Beijnen
Jos H
JH
D000911:Antibodies, Monoclonal / Q000032:analysis*; D061067:Antibodies, Monoclonal, Humanized; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D012680:Sensitivity and Specificity; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*; D000068878:Trastuzumab
D000911:Antibodies, Monoclonal; D061067:Antibodies, Monoclonal, Humanized; D000068878:Trastuzumab