Panax genus belonging to a family of Araliaceae grow in Asia(9 species)and in North America(2 species).Especially Panax ginseng was listed in Chinese medical book,Shennong's Classic of Materia Medica approximately...Panax genus belonging to a family of Araliaceae grow in Asia(9 species)and in North America(2 species).Especially Panax ginseng was listed in Chinese medical book,Shennong's Classic of Materia Medica approximately 2 thousand years ago and Panax species are now one of the most important natural medicine.Since Panax species contain approximately 260 ginsenosides,its quality control and pharmacological movement of ginsenoside in body are not enough understanding.Monoclonal antibodies against ginsenosides were prepared and set up the enzyme linked immunosorbent assay system for the quality control of natural product.Furthermore,we developed Eastern blotting system using monoclonal antibodies resulted that protopanaxadiol and protopanaxatriol group ginsenosides can be separately stained by two corresponding monoclonal antibodies,respectively.It became evident that the ginseng slice was stained by Eastern blotting system.Histochemical staining of ginseng can make clear the ginsenoside-Rb1 distribution in cells and tissues.Double Eastern blotting system facilitated by two monoclonal antibodies like anti-ginsenoside-Rb1 and anti-ginsenoside-Rg1 monoclonal antibodies gives several information such as sugar number,structure,and qualitative/quantitative evaluation.Immunoaffinity column combined with monoclonal antibodies succeeded one-step isolation of ginsenoside and make it possible to prepare knockout extract of which only antigen molecule was removed suggesting the pharmacological and biological value of antigen molecule in the crude extract.展开更多
Ginseng is well-known traditional medicine especially in Asian countries.Ginseng was listed in 120 safety medicines of the Chinese book,Shennong’s Classic of Materia Medica(written in the Eastern Han Dynasty;25–220 ...Ginseng is well-known traditional medicine especially in Asian countries.Ginseng was listed in 120 safety medicines of the Chinese book,Shennong’s Classic of Materia Medica(written in the Eastern Han Dynasty;25–220 C.E.).In Panax species several components like ginsenosides,triterpene saponins,polyacetylenes,polysaccharides,phenolics,alkaloids,lignans and peptides are contained^([1]).Yang et al.reviewed that 257 typical dammarane-type triterpenes,ginsenosides were isolated from Panax species and divided into protopanaxadiol and protopanaxatriol types having sugars attached to hydroxyl group at C-3 and C-20 or at C-6 and C-20,respectively^([2]).Ginsenoside Rc is a protopanaxadiol type ginsenoside such as a target compound taken up in the corresponding paper^([3]).展开更多
Green fluorescent protein (GFP) is a protein that emits green fluorescence when exposed to a radiation of ultraviolet wavelength range, even without the addition of substrate and cofactor. Because of such characterist...Green fluorescent protein (GFP) is a protein that emits green fluorescence when exposed to a radiation of ultraviolet wavelength range, even without the addition of substrate and cofactor. Because of such characteristics, the usage of GFP is widespread in both in vivo and in vitro applications. In addition, recent advances in biotechnology have enabled GFP to be expressed in various hosts, including bacteria, yeast, plants, animals, and even living-cells, for multiple purposes. Currently, GFP is a subject of great interest in the analytical sciences, especially in immunoassays for qualitative and quantitative analyses, when it is fused with an antibody because of the high sensitivity of GFP and antigen-binding specificity of antibodies. Recently the fluobody, which is a fusion protein of GFP with single-chain variable fragment antibody (scFv), has become a useful tool in various fields. We review here the applications of GFP as fluobodies in immunoassays.展开更多
We prepared many kinds of monoclonal antibodies(MAbs)against natural products and developed a new staining method using MAb named as Eastern blotting.Natural products containing glycoside were developed by TLC and the...We prepared many kinds of monoclonal antibodies(MAbs)against natural products and developed a new staining method using MAb named as Eastern blotting.Natural products containing glycoside were developed by TLC and the TLC plate was covered by PVDF or PES membrane and blotted.The membrane was treated with NaIO4,and展开更多
Monoclonal antibodies(MAbs) against natural products with low-molecular weights have become an important tool when combined with other analytical systems. Eastern blotting involves a typical staining system wherein, f...Monoclonal antibodies(MAbs) against natural products with low-molecular weights have become an important tool when combined with other analytical systems. Eastern blotting involves a typical staining system wherein, for example, glycosides can be blotted to a membrane and cross-linked and stained using MAbs. An immunoaffinity column combined with a monoclonal antibody allows a one-step purification of hapten compounds or preparation of a knockout extract that removes only the target hapten molecules from a crude extract. Here, we discuss the application of these extracts. Single-chain variable fragment(scFv) proteins have led to novel assay systems such as fluobodies or antibodies coupled with green fluorescent protein for natural products. A typical novel application of a scFv gene would be in the context of plant breeding; this is designated "missile-type" molecular breeding, intended to increase hapten molecule concentrations in transgenic plants.展开更多
文摘Panax genus belonging to a family of Araliaceae grow in Asia(9 species)and in North America(2 species).Especially Panax ginseng was listed in Chinese medical book,Shennong's Classic of Materia Medica approximately 2 thousand years ago and Panax species are now one of the most important natural medicine.Since Panax species contain approximately 260 ginsenosides,its quality control and pharmacological movement of ginsenoside in body are not enough understanding.Monoclonal antibodies against ginsenosides were prepared and set up the enzyme linked immunosorbent assay system for the quality control of natural product.Furthermore,we developed Eastern blotting system using monoclonal antibodies resulted that protopanaxadiol and protopanaxatriol group ginsenosides can be separately stained by two corresponding monoclonal antibodies,respectively.It became evident that the ginseng slice was stained by Eastern blotting system.Histochemical staining of ginseng can make clear the ginsenoside-Rb1 distribution in cells and tissues.Double Eastern blotting system facilitated by two monoclonal antibodies like anti-ginsenoside-Rb1 and anti-ginsenoside-Rg1 monoclonal antibodies gives several information such as sugar number,structure,and qualitative/quantitative evaluation.Immunoaffinity column combined with monoclonal antibodies succeeded one-step isolation of ginsenoside and make it possible to prepare knockout extract of which only antigen molecule was removed suggesting the pharmacological and biological value of antigen molecule in the crude extract.
文摘Ginseng is well-known traditional medicine especially in Asian countries.Ginseng was listed in 120 safety medicines of the Chinese book,Shennong’s Classic of Materia Medica(written in the Eastern Han Dynasty;25–220 C.E.).In Panax species several components like ginsenosides,triterpene saponins,polyacetylenes,polysaccharides,phenolics,alkaloids,lignans and peptides are contained^([1]).Yang et al.reviewed that 257 typical dammarane-type triterpenes,ginsenosides were isolated from Panax species and divided into protopanaxadiol and protopanaxatriol types having sugars attached to hydroxyl group at C-3 and C-20 or at C-6 and C-20,respectively^([2]).Ginsenoside Rc is a protopanaxadiol type ginsenoside such as a target compound taken up in the corresponding paper^([3]).
文摘Green fluorescent protein (GFP) is a protein that emits green fluorescence when exposed to a radiation of ultraviolet wavelength range, even without the addition of substrate and cofactor. Because of such characteristics, the usage of GFP is widespread in both in vivo and in vitro applications. In addition, recent advances in biotechnology have enabled GFP to be expressed in various hosts, including bacteria, yeast, plants, animals, and even living-cells, for multiple purposes. Currently, GFP is a subject of great interest in the analytical sciences, especially in immunoassays for qualitative and quantitative analyses, when it is fused with an antibody because of the high sensitivity of GFP and antigen-binding specificity of antibodies. Recently the fluobody, which is a fusion protein of GFP with single-chain variable fragment antibody (scFv), has become a useful tool in various fields. We review here the applications of GFP as fluobodies in immunoassays.
文摘We prepared many kinds of monoclonal antibodies(MAbs)against natural products and developed a new staining method using MAb named as Eastern blotting.Natural products containing glycoside were developed by TLC and the TLC plate was covered by PVDF or PES membrane and blotted.The membrane was treated with NaIO4,and
文摘Monoclonal antibodies(MAbs) against natural products with low-molecular weights have become an important tool when combined with other analytical systems. Eastern blotting involves a typical staining system wherein, for example, glycosides can be blotted to a membrane and cross-linked and stained using MAbs. An immunoaffinity column combined with a monoclonal antibody allows a one-step purification of hapten compounds or preparation of a knockout extract that removes only the target hapten molecules from a crude extract. Here, we discuss the application of these extracts. Single-chain variable fragment(scFv) proteins have led to novel assay systems such as fluobodies or antibodies coupled with green fluorescent protein for natural products. A typical novel application of a scFv gene would be in the context of plant breeding; this is designated "missile-type" molecular breeding, intended to increase hapten molecule concentrations in transgenic plants.
