title: Application of nano-surface and molecular-orientation limited proteolysis to LC-MS bioanalysis of cetuximab.
abstract: BACKGROUND
  We recently reported the principle of nano-surface and molecular-orientation limited (nSMOL) proteolysis, which is useful for LC-MS bioanalysis of antibody drugs.
  
  
  METHODS
  The nSMOL is a Fab-selective limited proteolysis which utilizes the difference of protease-immobilized nanoparticle diameter (200 nm) and antibody collection resin pore (100 nm). We have demonstrated the full validation for chimeric antibody cetuximab bioanalysis in human plasma using nSMOL. Signature peptides (SQVFFK, ASQSIGTNIHWYQQR and YASESISGIPSR) in cetuximab complementarity-determining region were simultaneously quantitated by LC-MS multiple reaction monitoring.
  
  
  CONCLUSIONS
  This nSMOL quantification showed sensitivity of 0.586 µg/ml and linearity of 0.586 to 300 µg/ml. Full validation study archived the guideline criteria of low Mw drug compounds. These results indicate that nSMOL is also significant method for cetuximab bioanalysis.
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: Megumi
authors.initials: M
authors.lastname: Takanashi
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: Division of Clinical Pharmacology & Translational Research, Exploratory Oncology Research & Clinical Trial Center, National Cancer Center, Tokyo, 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.
authors.forename: Takashi
authors.initials: T
authors.lastname: Shimada
mesh_terms: D000595:Amino Acid Sequence; D000970:Antineoplastic Agents / Q000032:analysis / Q000097:blood*; D000068818:Cetuximab / Q000032:analysis / Q000097:blood*; D002853:Chromatography, Liquid / Q000379:methods; D005260:Female; D006801:Humans; D055767:Immobilized Proteins / Q000737:chemistry; D057230:Limit of Detection; D008297:Male; D013058:Mass Spectrometry / Q000379:methods*; D008958:Models, Molecular; D049329:Nanostructures / Q000737:chemistry; D059748:Proteolysis; D016415:Sequence Alignment; D014357:Trypsin / Q000737:chemistry
chemical_list: D000970:Antineoplastic Agents; D055767:Immobilized Proteins; D014357:Trypsin; D000068818:Cetuximab
keywords: LC–MS bioanalysis; clinical pharmacokinetics; nSMOL