title: Sodium periodate-mediated conjugation of harringtonine enabling the production of a highly specific monoclonal antibody, and the development of a sensitive quantitative analysis method.
abstract: Harringtonine (HT) is a promising natural product that is mainly isolated from plants of the genus Cephalotaxus. Due to its remarkable antileukemic activities, HT has been utilized clinically in China for the treatment of acute promyelocytic leukemia (APL). No antibody that recognizes free HT has been reported to date due to the difficulty of preparing antigen conjugates in which haptens bind to a carrier protein. To overcome this difficulty, we focused on sodium periodate (NaIO4), which catalyzes unique oxidative reactions; the resulting conjugates enabled the production of a highly specific monoclonal antibody (MAb) against HT (MAb 1D2) and the establishment of an indirect competitive enzyme-linked immunosorbent assay (icELISA) for the determination of HT. Further analysis revealed that MAb 1D2 was produced by the HT3 (8-carbonyl HT)-based conjugate antigen; HT3 was synthesized by a NaIO4-mediated oxidative reaction. The minimum detectable concentration for HT in the icELISA system was found to be 0.76 ng mL-1, which is approximately 13 to 160 times more sensitive than a conventional HPLC system. Several validation analyses revealed that the icELISA using MAb 1D2 is sufficiently accurate, reliable, and sensitive to assess small amounts of HT in plant samples.
authors.affiliation: Graduate School of Pharmaceutical Sciences, Kyushu University, 3-1-1 Maidashi, Higashi-ku, Fukuoka 812-8582, Japan. s.sakamoto@phar.kyushu-u.ac.jp.
authors.forename: Seiichi
authors.identifier: http://orcid.org/0000-0001-6871-522X
authors.initials: S
authors.lastname: Sakamoto
authors.affiliation: Graduate School of Pharmaceutical Sciences, Kyushu University, 3-1-1 Maidashi, Higashi-ku, Fukuoka 812-8582, Japan. s.sakamoto@phar.kyushu-u.ac.jp.
authors.forename: Gorawit
authors.initials: G
authors.lastname: Yusakul
authors.affiliation: Graduate School of Pharmaceutical Sciences, Kyushu University, 3-1-1 Maidashi, Higashi-ku, Fukuoka 812-8582, Japan. s.sakamoto@phar.kyushu-u.ac.jp.
authors.forename: Yumi
authors.initials: Y
authors.lastname: Tsuneura
authors.affiliation: Department of Pharmacognosy, Faculty of Pharmaceutical Sciences, Khon Kaen University, Khon Kaen 40002, Thailand.
authors.forename: Waraporn
authors.initials: W
authors.lastname: Putalun
authors.affiliation: Graduate School of Pharmaceutical Sciences, Kyushu University, 3-1-1 Maidashi, Higashi-ku, Fukuoka 812-8582, Japan. s.sakamoto@phar.kyushu-u.ac.jp.
authors.forename: Kazuteru
authors.initials: K
authors.lastname: Usui
authors.affiliation: Graduate School of Pharmaceutical Sciences, Kyushu University, 3-1-1 Maidashi, Higashi-ku, Fukuoka 812-8582, Japan. s.sakamoto@phar.kyushu-u.ac.jp.
authors.forename: Tomofumi
authors.initials: T
authors.lastname: Miyamoto
authors.affiliation: Graduate School of Pharmaceutical Sciences, Kyushu University, 3-1-1 Maidashi, Higashi-ku, Fukuoka 812-8582, Japan. s.sakamoto@phar.kyushu-u.ac.jp.
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Tanaka
authors.affiliation: Graduate School of Pharmaceutical Sciences, Kyushu University, 3-1-1 Maidashi, Higashi-ku, Fukuoka 812-8582, Japan. s.sakamoto@phar.kyushu-u.ac.jp.
authors.forename: Satoshi
authors.initials: S
authors.lastname: Morimoto
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal; D032483:Cephalotaxus / Q000737:chemistry; D002851:Chromatography, High Pressure Liquid; D004797:Enzyme-Linked Immunosorbent Assay; D006248:Harringtonines / Q000737:chemistry*; D008297:Male; D008807:Mice, Inbred BALB C; D010504:Periodic Acid / Q000737:chemistry*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
chemical_list: D000911:Antibodies, Monoclonal; D006248:Harringtonines; D010504:Periodic Acid; C009288:metaperiodate