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Antibodies, Volume 1, Issue 3 (December 2012) – 4 articles , Pages 259-307

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Research

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607 KiB  
Article
Quality Control System for Beer Developed with Monoclonal Antibodies Specific to Barley Lipid Transfer Protein
by Yukie Murakami-Yamaguchi, Junko Hirose, Kumiko Kizu, Fumiko Okazaki, Wataru Fujii and Hiroshi Narita
Antibodies 2012, 1(3), 259-272; https://doi.org/10.3390/antib1030259 - 16 Oct 2012
Cited by 5 | Viewed by 7074
Abstract
Non-specific lipid transfer protein (LTP) in barley grain reacted with the IgE in sera drawn from food allergy patients. A sandwich-type of enzyme-linked immunosorbent assay (ELISA) was developed with mouse monoclonal antibodies raised against LTP purified with barley flour. This ELISA showed a [...] Read more.
Non-specific lipid transfer protein (LTP) in barley grain reacted with the IgE in sera drawn from food allergy patients. A sandwich-type of enzyme-linked immunosorbent assay (ELISA) was developed with mouse monoclonal antibodies raised against LTP purified with barley flour. This ELISA showed a practical working range of 0.3–3 ng/mL and no cross-reactivity with wheat, adlay and rye. Using this ELISA, LTP was determined in several types of barley-foods, including fermented foods such as malt vinegar, barley-malt miso and beer. LTP content in beer of the same kind was approximately constant, even if manufacturing factory and production days were different. Not only as a factor of foam formation and stability but also as an allergen, controlling and monitoring of LTP in beer should be considered. Taken together, our LTP-detecting ELISA can be proposed as an appropriate system for the quality control of beer. Full article
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634 KiB  
Article
Development of Eastern Blotting Technique for Analysis of Baicalin Using Anti-Baicalin Monoclonal Antibody
by Osamu Morinaga, Ryo Mukae, Takuhiro Uto, Yothawathorn Pariyawongsakul, Waraporn Putalun, Hiroyuki Tanaka and Yukihiro Shoyama
Antibodies 2012, 1(3), 284-293; https://doi.org/10.3390/antib1030284 - 27 Nov 2012
Cited by 3 | Viewed by 8244
Abstract
Scutellariae radix (S. radix) is one of the most important crude drugs used in Kampo medicines (KMs). A part of its pharmaceutical properties is due to flavone glycoside, baicalin (BI). A technique named eastern blotting was developed for the specific and [...] Read more.
Scutellariae radix (S. radix) is one of the most important crude drugs used in Kampo medicines (KMs). A part of its pharmaceutical properties is due to flavone glycoside, baicalin (BI). A technique named eastern blotting was developed for the specific and easy identification of BI in the extracts of crude drugs and KMs using anti-BI monoclonal antibody (MAb). BI separated by silica gel thin-layer chromatography (TLC) transferred to a polyethersulfone (PES) membrane was treated with a NaIO4 solution and reacted with bovine serum albumin (BSA) preparing BI-BSA conjugate on the PES membrane. Anti-BI MAb was bound and then antibody labeled with peroxidase directed against anti-BI MAb. Finally, a substrate was added and then BI was detected. As little as 1 mg of BI was still detected on the PES membrane under immunostaining method. Various samples of S. radix and KMs which contain S. radix were qualitatively analyzed, and BI was visually detected by eastern blotting technique. Furthermore, this method was applied for the immunohistochemical study to investigate the distribution of BI in the fresh root of Scutellaria baicalensis using immunoblotting by transferred from fresh root to the PES membrane. Full article
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Review

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249 KiB  
Review
Immunochemical Analysis of the Antimalarial Drugs Artemisinin and Artesunate
by Hiroyuki Tanaka, Madan K. Paudel, Ayako Takei, Junichi Sakoda, Thaweesak Juengwatanatrakul, Kaori Sasaki-Tabata, Waraporn Putalun, Wanchai De-Eknamkul, Oraphan Matangkasombut, Yukihiro Shoyama and Satoshi Morimoto
Antibodies 2012, 1(3), 273-283; https://doi.org/10.3390/antib1030273 - 02 Nov 2012
Cited by 1 | Viewed by 6798
Abstract
We prepared a monoclonal antibody (mAb 1C1) showing specificity for artemisinin (AM) and artesunate (AS), and we developed an indirect competitive enzyme-linked immunosorbent assay (icELISA) using this novel mAb. Moreover, we prepared a recombinant antibody derived from mAb 1C1 in order to overcome [...] Read more.
We prepared a monoclonal antibody (mAb 1C1) showing specificity for artemisinin (AM) and artesunate (AS), and we developed an indirect competitive enzyme-linked immunosorbent assay (icELISA) using this novel mAb. Moreover, we prepared a recombinant antibody derived from mAb 1C1 in order to overcome insufficient mAb production by hybridoma culture. A recombinant antigen-binding fragment (Fab) was easily constructed using antibody manipulation technologies and was produced by microorganisms in high yield. We herein review immunochemical approaches for analysis of the antimalarial drugs AM and AS that were able to yield analysis results for multiple samples in a short period of time using simple and reliable protocols. Full article
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355 KiB  
Review
Preparation of Knockout Extract by Immunoaffinity Column and Its Application
by Takuhiro Uto, Nguyen Huu Tung, Osamu Morinaga and Yukihiro Shoyama
Antibodies 2012, 1(3), 294-307; https://doi.org/10.3390/antib1030294 - 06 Dec 2012
Cited by 2 | Viewed by 6784
Abstract
Importance of herbal medicines have recently increased owing to rising interest in their health benefits. However, medicinal plant extracts are complex mixtures of phytochemicals that act synergistically or additively on specific and/or multiple molecular and cellular targets. Thus, it is difficult to examine [...] Read more.
Importance of herbal medicines have recently increased owing to rising interest in their health benefits. However, medicinal plant extracts are complex mixtures of phytochemicals that act synergistically or additively on specific and/or multiple molecular and cellular targets. Thus, it is difficult to examine the actual pharmacological roles of active compounds in plant extracts. This review describes a new strategy for isolating target compounds from plant extracts using immunoaffinity columns coupled with monoclonal antibodies (mAbs) against natural compounds. Through one-step purification using mAb-coupled immunoaffinity columns, we succeeded in preparing a knockout (KO) extract, which contains all components except the target compound. Furthermore, we investigated the pharmacological effects of the KO extract to reveal the actual effects of a bioactive compound in the crude extract. This approach may help determine the potential function of target compounds in herbal medicines. Full article
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