LC-MS/MS Bioanalysis of Trastuzumab Using Fab-Selective Proteolysis ‘nSMOL’

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One of the greatest benefits of antibody drugs is the safety as this same molecule constitutes our natural defence system against antigens. However, this benefit turns into a barrier to efficient bioanalysis; methodology is needed to selectively detect administered antibody without interference from host-derived antibodies. Although LC-MS/MS is expected to be the alternative to the conventional ELISA technique for its robustness and ease of method development, strong ionization suppression exerted by high-abundance peptides derived from the constant (Fc) region limits the sensitivity of low-abundance peptides in the variable (Fab) region.

Nano-surface and molecular orientation limited proteolysis (nSMOL™) is Shimadzu’s proprietary, innovative technology that enables selective proteolysis of the Fab region of antibodies. Peptide sample prepared by the nSMOL kit has substantially lower complexity near-zero interference from host antibodies, thus enabling LC-MS/MS sensitivity to reach down to the level required for pre-clinical and clinical studies. This application news reports the fully-validated LC-MS/MS method developed for Trastuzumab determination in plasma.

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