||Yusakul, G., Sakamoto, S., Tanaka, H., Morimoto, S., Modification of first constant domain of heavy chain (CH1) enabled effective folding of functional anti-forskolin antigen-binding fragment (Fab) for sensitive quantitative analysis., Biotechnol. Prog., in press, 2019.05.
||Yusakul, G., Sakamoto, S., Chanpokapaiboon, K., Tanaka, H., Morimoto, S, Preincubation format for a sensitive immunochromatographic assay for monocrotaline, a toxic pyrrolizidine alkaloid., Phytochem. Anal., in press, 2019.05.
||Sakamoto, S., Wada, S., Morita, Y., Yamaguchi, T., Tanaka, H., Morimoto, S., Magnetic particles-based enzyme immunoassay for rapid determination of secoiridoid glycoside, amarogentin, Talanta, in press, 2019.01.
||Sakamoto S.,Wada, S., Tanaka, H., Morimoto, S., Sensitive quantitative analysis of the bitter glycoside, amarogentin by specific monoclonal antibody-based indirect competitive enzyme-linked immunosorbent assay, RSC Advances, in press, 2018.05.
||＃Pongkitwitoon, B., Sakamoto, S., Nagamitsu, R., Putalun W., Tanaka, H., Morimoto, S., A monoclonal antibody-based enzyme-linked immunosorbent assay for determination of homoharringtonine, Planta Medica, in press, 2018.02.
||Sakamoto S, Miyamoto T, Usui K, Tanaka H, Morimoto S, Sodium-periodate Mediated Harringtonine Derivatives and Their Antiproliferative Activity against HL-60 Acute Leukemia Cells, Journal of Natural Products, 81, 1, 34-40, 2018.01, Harringtonine (HT) is a naturally occurring alkaloid isolated from the plant genus
Cephalotaxus. It possesses antileukemic activity and has been clinically utilized for the treatment of
acute leukemia and lymphoma. Sodium periodate (NaIO4) was reacted with HT to produce five HT
derivatives including four novel compounds. Their antiproliferative activity against HL-60 acute
promyelocytic leukemia cells revealed that the presence of the C-5′ methyl group enhances the
antiproliferative activity because the IC50 values of the HT derivatives, including HT1
(5′-de-O-methylharringtonine), was at least 2,000 times higher (> 100 μM) than that of HT (~47 nM).
In addition, an indirect competitive enzyme-linked immunosorbent assay (icELISA) using a
monoclonal antibody against HT (MAb 1D2) revealed that these antiproliferative activities were
related to their cellular uptake. These results indicated that esterification of HT1 at the C-4′
carboxylic acid group may enhance the antiproliferative activity of HT..
||Paudel, MK., Sakamoto, S., Tanaka, H., Morimoto, S., An overview and comparison of a recombinant antigen-binding fragment and an antigen-binding fragment from a monoclonal antibody against wogonin glucuronide, Journal of Natural Medicines, 71, 4, 703-710, 2017.10.
||Sakamoto S, Nagamitsu R, Yusakul G, Tomofumi M, Hiroyuki Tanaka, Morimoto S, Ultrasensitive immunoassay for monocrotaline using monoclonal antibody produced by N, N'-carbonyldiimidazole mediated hapten-carrier protein conjugates, Talanta, 168, 67-72, 2017.06.
||Paudel M, Sakamoto S, Huy LV, Hiroyuki Tanaka, Miyamoto T, Morimoto S, The effect of varying the peptide linker length in a single chain variable fragment antibody against wogonin glucuronide, Journal of Biotechnology, 251, 47-52, 2017.06.
||Sakamoto S, Yusakul G, Tsuneura Y, Putalun W, Usui K, Miyamoto T, Tanaka H, Morimoto S, Sodium periodate-mediated conjugation of harringtonine enabling the production of a highly specific monoclonal antibody, and the development of a sensitive quantitative analysis method, Analyst, 142, 1140-1148, 2017.04.
||Sakamoto S., Yusakul G., Nuntawong P., Kitisripanya T., Putalun W., Miyamoto T., Tanaka H., Morimoto S, Development of an indirect competitive immunochromatographic strip test for rapid detection and determination of anticancer drug, harringtonine, J. Chromatgr. B, 1048, 150-154, 2017.03.
||Paudel M, Sakamoto S, Huy LV, Hiroyuki Tanaka, Miyamoto T, Takano A, Morimoto S, Development of an immunoassay using an anti-wogonin glucuronide monoclonal antibody, J. Immunoassay Immunochem., 38, 2017.02.
||Taura F, Iijima M, Yamanaka E, Takahashi H, Saeki H, Morimoto S, Asakawa Y, Kurosaki F, Morita H, A novel class of plant type III polyketide synthase involved in orsellinic acid biosynthesis from Rhododendron dauricum, Frontiers in Plant Science, 7, 記事番号1452, 2016.09.
||Yusakul G., Sakamoto S, Pongkitwitoon B, Tanaka H, Morimoto S, Effect of linker length between variable domains of single chain variable fragment antibody against daidzin on its reactivity, Biosci. Biotechnol. Biochem., 80, 1306-1312, 2016.07.
||Yusakul G, Sakamoto S, Tanaka H, Morimoto S, Efficient expression of single chain variable fragment antibody against paclitaxel using Bombyx mori nucleopolyhedrovirus bacmid DNA system and its characterizations, J. Nat. Med., 70, 3, 592-601, 2016.07.
||Sakamoto S, Kohno T, kuniyoshi shimizu, Tanaka H, Morimoto S, Detection of ganoderic acid A in Ganoderma lingzhi by an indirect competitive enzyme-linked immunosorbent assay, Planta Medica, 82, 747-751, 2016.05.
||Sakamoto S, Yusakul G, Pongkitwitoon B, Tanaka H, Morimoto S, Colloidal gold-based indirect competitive immunochromatographicassay for rapid detection of bioactive isoflavone glycosides daidzin andgenistin in soy products, Food Chem., 194, 191-195, 2016.03.
||Yusakul G, Morimoto S, Juengwatanatrakul T, Putalun W, Tanaka H, Sakamoto S, Preparation and application of a monoclonal antibody against the isoflavone glycoside daidzin using a Mannich reaction-derived hapten conjugate, Phytochem. Anal., 27, 81-85, 2016.01.
||Sakamoto S, Nagamitsu R, Matsuura Y, Tsuneura Y, Kurose H, Tanaka H, Morimoto S, A new approach of indirect enzyme-linked immunosorbent assay for determination of D-glutamic acid through in situ conjugation, Journal of Immunoassay and Immunochemistry, in press, 2016.01.
||Yusakul G, Udomsin O, Morimoto S, Tanaka H, Juengwatanatrakul T, Putalun W, Enzyme-linked immunosorbent assay by enhanced chemiluminescence detection for the standardization of estrogenic miroestrol in Pueraria candollei Graham ex Benth, Luminescence, 30, 568-575, 2015.08.
||Seiichi Sakamoto, Yusakul G., Pongkitwitoon B, Paudel, M.K., Hiroyuki Tanaka, Satoshi Morimoto, “Simultaneous determination of soy isoflavone glycosides, daidzin and genistin by monoclonal antibody-based highly sensitive indirect competitive enzyme-linked immunosorbent assay, Food. Chem., 169, 127-133, 2015.01.
||Limsuwanchote S, Wungsintaweekul J, Yusakul G, Han J.Y., Kaori Tabata, Hiroyuki Tanaka, Yukihiro Shoyama, Satoshi Morimoto, Preparation of a monoclonal antibody against notoginsenoside R1, a distinctive saponin fromPanax notoginseng, and its application to indirect competitive ELISA, Planta Med, 80, 4, 337-342, 2014.04.
||Tanaka H., Paudel M., Juengwatanatrakul T., Sasaki-Tabata K., Waraporn P., De-Eknamkul W., Matangkasombut O., Shoyama Y., Morimoto, S., Fluobodies against Bioactive Natural Product and Its Application in Fluorescence-Linked Immunosorbent Assay
, Antibodies, in press, 2012.10, Immunochemical Analysis of Anti-malarial Drugs, Artemisinin and Artesunate.
||Yoshinari Shoyama, Taro Tamada, Kazuo Kurihara, Ayako Takeuchi, Futoshi Taura, Shigeki Arai, Blaber Michael, Yukihiro Shoyama, Satoshi Morimoto,
Ryota Kuroki, Ph.D.
, Tetrahydrocannabinolic Acid (THCA) Synthase, the Enzyme Controlling the Psychoactivity of Cannabis sativa
, Journal of Molecular Biology, 423, 96-105, 2012.10, ∆1-Tetrahydrocannabinolic acid (THCA) synthase catalyzes the oxidative cyclization of cannabigerolic acid (CBGA) into THCA, the precursor of the primary psychoactive agent ∆1-tetrahydrocannabinol in Cannabis sativa. In order to investigate the structure-function relationship of THCA synthase, the enzyme was over-produced in insect cells, purified, crystallized, and the tertiary structure determined to 2.75 Å resolution by X-ray crystallography (Rcryst=19.9%). The THCA synthase enzyme is a member of the p-cresol methyl-hydroxylase superfamily, and the tertiary structure is divided into two domains (domains I and II), with a flavin adenine dinucleotide (FAD) coenzyme positioned between each domain and covalently bound to His114 and Cys176 (located in domain I). The ionized residues in the active site of THCA synthase were investigated by mutational analysis and X-ray structure. The catalysis of THCA synthesis involves a hydride transfer from C3 of CBGA to N5 of flavin adenine dinucleotide (FAD) and the deprotonation of O6’ of CBGA. Mutation analysis indicates that the reaction does not involve the carboxyl group of Glu442 that was identified as the catalytic base in the related Berberine Bridge enzyme (BBE), but instead involves the hydroxyl group of Tyr484. Mutations at the active site residues His292 and Tyr417 resulted in a decrease in, but not elimination of, the enzymatic activity of THCA synthase, suggesting a key role for these residues in substrate binding and not direct catalysis. .
||Sakamoto S, Pongkitwitoon B, Nakahara H, Shibata O, Shoyama Y, Tanaka H, Morimoto S
, Fluobodies against Bioactive Natural Product and Its Application in Fluorescence-Linked Immunosorbent Assay
, Antibodies, 1, 239-258, 2012.09, Abstract: An enzyme-linked immunosorbent assay (ELISA) using monoclonal antibody (MAb), Fab antibody, and single-chain variable fragment (scFv) antibody have become one of the promising analytical methods owing to its rapidity, sensitivity, and reliability. Recently, a chimera of green fluorescent protein (GFP) with a scFv antibody, named fluobody, has been proposed as a probe for an alternative immunosorbent assay; i.e., fluorescence-linked immunosorbent assay (FLISA). In this FLISA, even more sensitive, simple, and rapid immunoassay can be performed by detecting highly sensitive fluorophore of GFP that is genetically and directly fused to scFv antibody. In addition, time- and cost-consuming secondary antibody reaction and following enzyme-substrate reaction necessary for conventional ELISA can be avoided, making it possible to complete the assay more rapid. Focusing on naturally occurring bioactive product, fluobody recognizing 1,4-naphthoquinone, plumbagin and triterpenoid saponin, ginsenosides was successfully expressed in Escherichia coli (E. coli) and applied them to FLISA. The construction, expression, and the potential use of fluobody in quantitative/qualitative analysis of bioactive natural product have been reviewed in this article.
||Sritularak, B, Juengwatanatrakul, T, Putalun, W, Tanaka, H, Morimoto, S , A rapid one-step immunochromatographic assay for the detection of asiaticoside, Journal of Natural Medicines, 10.1007/s11418-011-0582-2, 78, 11, 1287-1288, 2012.04, Asiaticoside has been identified as the most active compound in Centella asiatica. In order to screen a large number of plant samples for the presence of asiaticoside, a rapid and simple technique is required that utilizes small quantities for test samples. In this study, an immunochromatographic strip test has been developed for the detection of asiaticoside in plant samples that uses a monoclonal antibody against asiaticoside. The limit of detection for the strip test was 12.5 mu g/ml. Immunoassay using monoclonal antibodies could be useful for the determination of small quantities of asiaticoside in plant extracts. .
||Paudel M.K., Putalun W., Sritularak B, Morinaga, O., Shoyama Y., Tanaka H. and Morimoto S., Development pf a combined technique using a rapid one-step immunochromatographic assay and indirect competitive ELISA for the rapid detection of bacalin, Analytica Chimica Acta, 10.1016/j.aca.2011.05.054 , 701, 2, 189-193, 701, (2) 189-193, 2012.02, A colloidal gold conjugated anti-baicalin monoclonal antibody (anti-BA MAb) was prepared and used in an immunochromatographic assay (ICA) for BA in Scutellariae Radix and Kampo medicines. This competitive ICA uses an anti-BA MAb which shows a high specificity for BA and baicalein. Its advantages include a short assay time (15 min), no dependence on any instrumental systems, and it can detect BA in plant materials and Kampo medicines. The limit of detection for the ICA was found to be around 0.6 mu g mL(-1) of baicalin. Moreover, the usefulness of the combination of indirect competitive ELISA and the ICA using anti-BA MAb as a quality control method was confirmed for analysis of BA in Scutellariae Radix and Kampo medicines with a sufficient sensitivity (200 ng mL(-1) to 2 mu g mL(-1)), obtainable in an easy and timely manner. .
||Madan K Paudel, Ayako Takei, Junichi Sakoda, Juengwatanatrakul, Thaweesak, Kaori Tabata, Putalun Waraporn, Shoyama Yukihiro, Hiroyuki Tanaka, Satoshi Mrimoto, Preparation of a single-chain variable fragment and a recombinant antigen-binding fragment against the anti-malarial drugs, artemisinin and artesunate, and their application in an ELISA, Analytical Chemitry, 10.1021/ac203131f, 701, 2, 2002-2008, 2012.02, Two different recombinant antibodies, a single-chain variable fragment (scFv) and an antigen-binding fragment (Fab), were prepared against artemisinin (AM) and artesunate (AS) and were developed for use in an enzyme-linked immunosorbent assay (ELISA). The recombinant antibodies, which were derived from a single monoclonal antibody against AM and AS (mAb 1C1) prepared by us, were expressed by Escherichia coli cells and their reactivity and specificity were characterized. As a result, to obtain sufficient signal in indirect ELISA, a much greater amount of a first antibody was needed in the use of scFv due to the differences of the secondary antibody and conformational stability. Therefore, we focused on the development of the recombinant Fab antibodies and applied it to indirect competitive ELISA. The specificity of the Fab was similar to that of mAb 1C1 in that it showed specific reactivity toward AM and AS only. The sensitivity of the icELISA (0.16 mu g/mL to 40 mu g/mL for AM and 8.0 ng/mL to 60 ng/mL for AS) was sufficient for analysis of antimalarial drugs, and its utility for quality control of analysis of Artemisia spp. was validated. The Fab expression and refolding systems provided a good yield of high-quality antibodies. The recombinant antibody against AM and AS provides an essential component of an economically attractive immunoassay and will be useful in other immunochemical applications for the analysis and purification of antimalarial drugs.iae Radix and Kampo medicines with a sufficient sensitivity (200 ng mL(-1) to 2 mu g mL(-1)), obtainable in an easy and timely manner. .
||Sakamoto, S., Putalun, W., Pongkitwitoon, B., Shoyama, Y., Tanaka, H., Morimoto, S., Modulation of plumbagin production in Plumbago zeylanica using a single-chain variable fragment antibody against plumbagin, Plant Cell Reports, 10.1007/s00299-011-1143-6, 31, 1, 103-110, 2012.01, A single-chain variable fragment antibody (scFv) against plumbagin (PL) accumulated the PL production in the hairy roots of Plumbago zeylanica. Recombinant Agrobacterium rhizogenes (ATCC 15834) containing an scFv gene against PL (PL-scFv) were obtained through triparental mating and transformed into P. zeylanica to induce PL-scFv protein in the hairy roots. Up to 40 mu g recombinant PL-scFv were expressed per milligram of soluble protein in transgenic P. zeylanica hairy root cultures. The mean PL content obtained from transgenic hairy roots (12.24 mu g/100 mg dry weight) exhibited 2.2 times higher than those obtained from wild-type (5.48 mu g/100 mg dry weight). The high correlation between the PL-scFv expression level and PL content of the recombinant plants suggested that the PL biosynthesis pathway had been modulated by the expression of PL-scFv protein in the hairy roots of P. zeylanica. .
||Sakamoto, S., Pongkitwitoon, B., Sasaki-Tabata, K., Putalun, W., Maenaka, K., Tanaka, H., Morimoto, S.
, A fluorescent single domain antibody against plumbagin expressed in silkworm larvae for fluorescence-linked immunosorbent assay (FLISA)
, Analyst, 10.1039/c1an15027h , 136, 10, 2056-2063, 136 (10) 2056-2063, 2011.10.
||Motosuke Hirunuma, Yoshinari Shoyama, Kaori Sasaki, Seiichi Sakamoto, Futoshi Taura, Yukihiro Shoyama, Hiroyuki Tanaka and Satoshi Morimoto, Flavone-catalyzed apoptosis in Scutellaria baicalensis, Phytochemistry, 10.1016/j.phytochem.2011.02.009 , 72, 8, 752-760, 72 (8) 752-760, 2011.06.
||Sakamoto, S., Pongkitwitoon, B., Nakamura, S., Sasaki-Tabata, K., Tanizaki, Y., Maenaka, K. Tanaka, H., Morimoto, S.
, Construction, expression, and characterization of a single-chain variable fragment antibody against 2, 4- dichlorophenoxyacetic acid in the hemolymph of silkworm larvae, Appl. Biochem. Biotechnol., 10.1007/s12010-011-9168-4 , 164, 6, 715-728, 164 (6) 715-728, 2011.06.
||Sakamoto, S., Tanizaki, Y., Pongkitwitoon, B., Tanaka, H., Morimoto, S., A chimera of green fluorescent protein with single chain variable fragment antibody against ginsenosides for fluorescence-linked immunosorbent assay
, Protein Expression and Purification, 10.1016/j.pep.2011.01.010 , 77, 1, 124-130 , 77 (1) 124-130, 2011.05.
||Juengwatanatraku, T., Sritularak, B., Amornnopparattanakul, P.,Tassanawat, P.,Putalun, W., Tanaka, H., Morimoto, S. , Preparation of a specific monoclonal antibody to asiaticoside for the development of an enzyme-linked immunosorbent assay , Analyst, 10.1039/c0an00868k , 136, 5, 1013-1017, 136 (5) 1013-1017, 2011.05.
||B. Pongkitwitoon, S. Sakamoto, O. Morinaga, T. Juengwatanatrakul, Y. Shoyama, H. Tanaka, S. Morimoto., Single-chain variable fragment antibody against ginsenoside Re as an effective tool for the determination of ginsenosides in various ginsengs, J. Nat. Med., 10.1007/s11418-010-0446-1 , 65, 1, 24-30, 65 (1) 24-30, 2011.01.
||S. Sakamoto, F. Taura, R. Tsuchihashi, W. Putalun, J. Kinjo, H. Tanaka, S. Morimoto, Expression, purification and characterization of anti-plumbagin single-chain variable fragment antibody in Sf9 insect cell, Hybridoma, 10.1089/hyb.2010.0052 , 29, 6, 481-488 , 29 (6) 481-488, 2010.12.
||S. Sakamoto, B. Pongkitwitoon, S. Nakamura, K. Maenaka, H. Tanaka, S. Morimoto, Efficient silkworm expression of single-chain variable fragment antibody against ginsenoside Re using Bombyx mori nucleopolyhedrovirus bacmid DNA system and its application in enzyme-linked immunosorbent assay for quality control of total ginsenosides”, Journal of Biochemistry, 10.1093/jb/mvq072 , 148, 3, 335-340 , 148 (3) 335-340 , 2010.09.
||S. Sakamoto, F. Taura, B. Pongkitwitoon, W. Putalun, R. Tsuchihashi, J. Kinjo, H. Tanaka, S. Morimoto.
, Development of sensitivity-improved fluorescence-linked immunosorbent assay using a fluorescent single domain antibody against bioactive naphthoquinone, plumbagin, Anal. Bioanal. Chem., 10.1007/s00216-010-3535-9 , 396, 8, 2955-2963, 396 (8) 2955-2963, 2010.04.
||F.Taura, S. Tanaka, C. Taguchi, T. Fukamizu, H. Tanaka, Y. Shoyama and S. Morimoto, Characterization of olivetol synthase, a polyketide synthase putatively involved in cannabinoid biosynthetic pathway, FEBS Letters, 10.1016/j.febslet.2009.05.024 , 583, 12, 2061-2066, 583 (12) 2061-2066, 2009.06.
||Yoshinari Shoyama, Chitomi Sugawa, Hiroyuki Tanaka, Satoshi Morimoto, Cannabinoids act as necrosis-inducing factors in Cannabis sativa, Plant Signaling & Behavior, 3, 12, 1111-1112, 3 (12) 1111-1112, 2008.12.
||Chiho Taguchi, Futoshi Taura, Taro Tamada, Yoshinari Shoyama, Yukihiro Shoyama, Hiroyuki Tanaka, Ryota Kuroki and Satoshi Morimoto, Crystallization and preliminary X-ray diffraction studies of polyketide synthase-1 (PKS-1) from Cannabis sativa, Acta Cryst. F, 10.1107/S1744309108003795 , f64, 3, 217-220, F64 (3) 217-220, 2008.02.
||F.Taura, E. Dono, S. Sirikantaramas, K. Yoshimura, Y. Shoyama, and S. Morimoto, Production of THCA by the biosynthetic enzyme secreted from transgenic Pichia pastoris, Biochem. Biophys. Res. Commun. , 10.1016/j.bbrc.2007.07.079, 361, 3, 675-680, 361(3) 675-680
||S. Morimoto, Y.Tanaka, K.Sasaki, H. Tanaka, T. Fukamizu, Y. Shoyama, Y. Shoyama and F.Taura, Identification and characterization of cannabinoids that induce cell death through mitochondrial permeablity transition in Cannabis leaf cells, J. Biol. Chem., 10.1074/jbc.M700133200 , 282, 28, 20739-20751 , 282 (28) 20739-20751, 2007.07.
||Sirikantaramas S, Morimoto S, Shoyama Y, Ishikawa Y, Wada Y, Shoyama Y, Taura F, The gene controlling marijuana psychoactivity molecular cloning and heterologous expression of D1-THCA from Cannabis sativa, J. Biol. Chem., 10.1074/jbc.M403693200, 279, 38, 39767-39774, 279 (38): 39767-39774 SEP 17 2004, 2004.09.