title: Enantioselective immunorecognition of protein modification with optically active ibuprofen using polyclonal antibody.
abstract: Formation of covalently bound protein adducts with 2-arylpropionic acids (2-APAs) has been proposed as a possible explanation for hypersensitivity and toxic responses to chiral carboxylic acid drugs. To identify the cellular proteins chemically modified with optically active (S)-ibuprofen, we generate polyclonal antibodies by immunizing rabbits with immunogen coupled to bovine serum albumin (BSA) via the spacer of 4-aminobutyric acid. The resulting antibodies largely cross-reacted with N-alpha-(t-butoxycarbonyl)--(S)-ibuprofenyl lysine as well as with the conjuguated (S)-ibuprofen with glycine and taurine and unconjugated (S)-ibuprofen, enabling enantioselective detection of (S)-ibuprofen residues anchored on ovalbumin molecules, introduced by the reaction of the ibuprofen p-nitrophenyl ester. Furthermore, immunoblotting with an antibody allows the enantioselective detection of (S)-ibuprofen-introduced glutathione-S-transferase (GST). These results indicate that the developed method will be useful for monitoring the generation and localization of protein covalently bound with (S)-ibuprofen, which may be the cause of ibuprofen-induced toxicity.
authors.affiliation: Faculty of Pharmaceutical Sciences, Kinki University, 3-4-1 Kowakae, Higashi-osaka 577-8502, Japan.
authors.forename: Hiromi
authors.initials: H
authors.lastname: Ito
authors.forename: Shunji
authors.initials: S
authors.lastname: Ishiwata
authors.forename: Takeshi
authors.initials: T
authors.lastname: Kosaka
authors.forename: Rika
authors.initials: R
authors.lastname: Nakashima
authors.forename: Harunori
authors.initials: H
authors.lastname: Takeshita
authors.forename: Sakiko
authors.initials: S
authors.lastname: Negoro
authors.forename: Masako
authors.initials: M
authors.lastname: Maeda
authors.forename: Shigeo
authors.initials: S
authors.lastname: Ikegawa
mesh_terms: D000595:Amino Acid Sequence; D000906:Antibodies / Q000276:immunology*; D004591:Electrophoresis, Polyacrylamide Gel; D007052:Ibuprofen / Q000737:chemistry* / Q000276:immunology; D009682:Magnetic Resonance Spectroscopy; D008969:Molecular Sequence Data; D011506:Proteins / Q000737:chemistry*; D013237:Stereoisomerism
chemical_list: D000906:Antibodies; D011506:Proteins; D007052:Ibuprofen