title: Fully validated LCMS bioanalysis of Bevacizumab in human plasma using nano-surface and molecular-orientation limited (nSMOL) proteolysis.
abstract: The chemistry of nano-surface and molecular-orientation limited (nSMOL) proteolysis is the Fab-selective limited proteolysis by making use the difference of protease nanoparticle diameter (200 nm) and antibody resin pore diameter (100 nm). In this report, we have demonstrated that the full validation for Bevacizumab bioanalysis in human plasma using nSMOL. The immunoglobulin fraction was collected by Protein A resin from plasma, then nSMOL reaction was performed using the FG nanoparticle-immobilized trypsin under the nondenaturing physiological condition at 50 °C for 6 h. After removal of resin and nanoparticles, the signature peptide of Bevacizumab complementarity-determining region (CDR) and internal standard P14R were simultaneously quantified by LCMS multiple reaction monitoring (MRM). This nSMOL method quantification of Bevacizumab showed sensitivity of 0.146 μg/ml and linearity of 0.146-300 μg/ml. The intra- and inter-assay precision of lower limit of quantification (LLOQ), low quality control (LQC), middle quality control (MQC), and high quality control (HQC) was 7.94-15.2% and 14.6%, 7.15-13.5% and 11.7%, 2.63-6.47% and 5.83%, and 3.09-4.35% and 4.45%, respectively. These results indicate that nSMOL is also significant method for Bevacizumab bioanalysis in human plasma.
authors.affiliation: Life Science Research Center, SHIMADZU Corporation, Kanda-Nishiki-cho, Chiyoda-ku, Tokyo 101-8448, Japan.
authors.forename: Noriko
authors.initials: N
authors.lastname: Iwamoto
authors.affiliation: Life Science Research Center, SHIMADZU Corporation, Kanda-Nishiki-cho, Chiyoda-ku, Tokyo 101-8448, Japan.
authors.forename: Yukari
authors.initials: Y
authors.lastname: Umino
authors.affiliation: Life Science Research Center, SHIMADZU Corporation, Kanda-Nishiki-cho, Chiyoda-ku, Tokyo 101-8448, Japan.
authors.forename: Chikage
authors.initials: C
authors.lastname: Aoki
authors.affiliation: CMIC Pharma Science Co., Ltd., Shibaura, Minato-ku, Tokyo 105-0023, Japan.
authors.forename: Naoe
authors.initials: N
authors.lastname: Yamane
authors.affiliation: Division of Clinical Pharmacology, National Cancer Center, Tsukiji, Chuo-ku, Tokyo 104-0045, Japan.
authors.forename: Akinobu
authors.initials: A
authors.lastname: Hamada
authors.affiliation: Life Science Research Center, SHIMADZU Corporation, Kanda-Nishiki-cho, Chiyoda-ku, Tokyo 101-8448, Japan. Electronic address: t-shima@shimadzu.co.jp.
authors.forename: Takashi
authors.initials: T
authors.lastname: Shimada
mesh_terms: D000068258:Bevacizumab / Q000097:blood* / Q000737:chemistry*; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D006801:Humans; D049329:Nanostructures / Q000737:chemistry*; D010949:Plasma / Q000737:chemistry*; D059748:Proteolysis; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D053719:Tandem Mass Spectrometry / Q000379:methods; D013810:Therapeutic Equivalency
chemical_list: D000068258:Bevacizumab
keywords: Antibody drugs; Bioanalysis; Clinical pharmacokinetics; LCMS; nSMOL