Issue 7, 2017

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.

Graphical abstract: Sodium periodate-mediated conjugation of harringtonine enabling the production of a highly specific monoclonal antibody, and the development of a sensitive quantitative analysis method

Article information

Article type
Paper
Submitted
30 Dec 2016
Accepted
11 Mar 2017
First published
13 Mar 2017

Analyst, 2017,142, 1140-1148

Sodium periodate-mediated conjugation of harringtonine enabling the production of a highly specific monoclonal antibody, and the development of a sensitive quantitative analysis method

S. Sakamoto, G. Yusakul, Y. Tsuneura, W. Putalun, K. Usui, T. Miyamoto, H. Tanaka and S. Morimoto, Analyst, 2017, 142, 1140 DOI: 10.1039/C6AN02751B

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