Fingerprinting of natural product by eastern blotting using monoclonal antibodies

2016 ◽  
Vol 07 (05) ◽  
Author(s):  
Yukihiro Shoyama
Keyword(s):  
Monoclonal Antibodies ◽  
Natural Product ◽  
Eastern Blotting

scholarly journals Fingerprinting of Natural Product by Eastern Blotting Using Monoclonal Antibodies

2012 ◽  
Vol 2012 ◽  
pp. 1-7
Author(s):  
Hiroyuki Tanaka ◽  
Waraporn Putalun ◽  
Yukihiro Shoyama
Keyword(s):  
Monoclonal Antibodies ◽  
Natural Product ◽  
Cross Reactivity ◽  
Blue Color ◽  
Ginsenoside Rb1 ◽  
Double Staining ◽  
Eastern Blotting ◽  
The Central Nervous System ◽  
Rf Values ◽  
Solasodine Glycosides

We succeeded in developing the fingerprint of natural product by eastern blotting using monoclonal antibodies. After developing and separating them on a TLC plate, solasodine glycosides are oxidized by NaIO4 and reacted with a protein to give conjugates which are recognized with anti-solamargine monoclonal antibody (MAb). Anti-solamargine MAb having wide cross-reactivity can stain and detect all solasodine glycosides by fingerprint. Different sensitivity between solamargine and solasonine was observed. The detection limit was 1.6 ng of solasonine. The hydrolysed products of solamargine were determined by fingerprint of eastern blotting compared to their Rf values depending on the sugar number. Fingerprint by eastern blotting using anti-ginsenoside Rb1 MAb distinguished the formula containing ginseng prescribed in traditional Chinese medicine. By double-staining of ginsenosides it is possible to suggest that the staining color shows the pharmacological activity, such as the purple bands indicate ginsenosides having stimulation activity, and the blue color indicated compound like ginsenosides possessed the depression affect for the central nervous system (CNS), respectively.

scholarly journals Studies on Natural Products Using Monoclonal Antibodies: A Review

Antibodies ◽  
2021 ◽  
Vol 10 (4) ◽  
pp. 43
Author(s):  
Yukihiro Shoyama
Keyword(s):  
Natural Products ◽  
Monoclonal Antibodies ◽  
Thin Layer ◽  
Small Molecule ◽  
Polyvinylidene Fluoride ◽  
Herbal Medicines ◽  
Sugar Moiety ◽  
Eastern Blotting ◽  
Tlc Plates ◽  
Layer Chromatography

An immunoblotting system (“eastern blotting”) was developed for small-molecule herbal medicines like glycosides, with no conjugation function to the membrane. Briefly, the crude extracts of herb medicines were developed by thin-layer chromatography (TLC). The small-molecule herbal medicines on TLC plates were transferred to polyvinylidene fluoride (PVDF) or polyethersulfone (PES) membranes by heating. Antigen components were divided into two categories based on their function, i.e., their membrane recognizing (aglycone part) and fixing (sugar moiety) abilities. This procedure allows for the staining of only target glycosides. Double eastern blotting was developed as a further staining system for two herb medicines using a set of MAbs and substrates.

NIH assistance for new therapeutic development: NIH-RAID Pilot Program

2009 ◽  
Vol 15 (1) ◽  
Author(s):  
David G Badman
Keyword(s):  
Gene Therapy ◽  
Monoclonal Antibodies ◽  
Natural Product ◽  
Small Molecule ◽  
Recombinant Proteins ◽  
Therapeutic Agents ◽  
Preclinical Development ◽  
Non Profit ◽  
Data Services ◽  
Therapeutic Development

The National Institutes of Health (NIH) wishes to alert the biotech/medical research community to an opportunity to obtain assistance in the development of new therapeutic agents. The NIH Roadmap has established a pilot programme, the NIH-Rapid Access to Interventional Development (RAID) Pilot, to make available, on a competitive basis, critical resources needed for the development of new small-molecule or natural product-derived therapeutic agents. This programme, part of the Translational Research component of Reengineering the Clinical Research Enterprise, uses resources of NCI's Developmental Therapeutics Program. Services provided depend upon the project and strength of the preliminary data. Services potentially available include bulk supply, GMP manufacturing, formulation, assay development suitable for pharmacokinetic testing, and animal toxicology. Assistance can also be provided in the regulatory process. Currently, animal efficacy studies and synthesis of recombinant proteins, monoclonal antibodies, or reagents for gene therapy are not supported. The NIH-RAID Pilot will, however, consider requests for services to support later-stage preclinical development of monoclonal antibodies, recombinant proteins, and gene therapy agents. Additionally, the NIH-RAID Pilot will now consider requests for the manufacture of small-molecule or natural product material for any clinical study. Proposals must originate from academic or non-profit investigators, but collaboration with industry partners is encouraged.

scholarly journals Pharmacologically Active Saikosaponin inBupleurum falcatumDetected by Competitive ELISA and Eastern Blotting Using Monoclonal Antibodies

2008 ◽  
Vol 11 (2) ◽  
pp. 192-197 ◽  
Author(s):  
Tae Kwon Shon ◽  
Shu-hang Zhu ◽  
Sang Chul Lee ◽  
Yukihiro Shoyama ◽  
Hiroyuki Tanaka
Keyword(s):  
Monoclonal Antibodies ◽  
Competitive Elisa ◽  
Eastern Blotting ◽  
Pharmacologically Active

Eastern blotting and use of anti-saikosaponin a monoclonal antibodies for detection of saikosaponins

2006 ◽  
Vol 61 (2) ◽  
pp. 178-183 ◽  
Author(s):  
Shu-hang Zhu ◽  
Osamu Morinaga ◽  
Shinichi Shimokawa ◽  
Tae Kwon Shon ◽  
Sang Chul Lee ◽  
...  
Keyword(s):  
Monoclonal Antibodies ◽  
Eastern Blotting ◽  
Saikosaponin A

Development of eastern blotting technique for sennoside A and sennoside B using anti-sennoside A and anti-sennoside B monoclonal antibodies

2009 ◽  
Vol 20 (2) ◽  
pp. 154-158 ◽  
Author(s):  
Osamu Morinaga ◽  
Takuhiro Uto ◽  
Seiichi Sakamoto ◽  
Waraporn Putalun ◽  
Sorasak Lhieochaiphant ◽  
...  
Keyword(s):  
Monoclonal Antibodies ◽  
Eastern Blotting ◽  
Blotting Technique

Ultrastructural analysis of unique glycoconjugates in the rat olfactory system

1992 ◽  
Vol 50 (1) ◽  
pp. 890-891
Author(s):  
James E. Crandall ◽  
Linda C. Hassinger ◽  
Gerald A. Schwarting
Keyword(s):  
Central Nervous System ◽  
Nervous System ◽  
Monoclonal Antibodies ◽  
Olfactory System ◽  
Olfactory Bulbs ◽  
Temporal And Spatial Patterns ◽  
Carbohydrate Epitopes ◽  
Olfactory Systems ◽  
Temporal And Spatial ◽  
Cell Surface Glycoconjugates

Cell surface glycoconjugates are considered to play important roles in cell-cell interactions in the developing central nervous system. We have previously described a group of monoclonal antibodies that recognize defined carbohydrate epitopes and reveal unique temporal and spatial patterns of immunoreactivity in the developing main and accessory olfactory systems in rats. Antibody CC2 reacts with complex α-galactosyl and α-fucosyl glycoproteins and glycolipids. Antibody CC1 reacts with terminal N-acetyl galactosamine residues of globoside-like glycolipids. Antibody 1B2 reacts with β-galactosyl glycolipids and glycoproteins. Our light microscopic data suggest that these antigens may be located on the surfaces of axons of the vomeronasal and olfactory nerves as well as on some of their target neurons in the main and accessory olfactory bulbs.

Structural and Chemical Studies of Pathological Fibers in Human Neurofibrillary Degeneration

1985 ◽  
Vol 43 ◽  
pp. 726-729
Author(s):  
K.S. Kosik ◽  
L.K. Duffy ◽  
S. Bakalis ◽  
C. Abraham ◽  
D.J. Selkoe
Keyword(s):  
Monoclonal Antibodies ◽  
Alzheimer Disease ◽  
Human Brain ◽  
Neurofibrillary Tangles ◽  
Morphological Analysis ◽  
High Specificity ◽  
Cytoskeletal Proteins ◽  
Neuritic Plaque ◽  
Protein Chemistry ◽  
Chemical Studies

The major structural lesions of the human brain during aging and in Alzheimer disease (AD) are the neurofibrillary tangles (NFT) and the senile (neuritic) plaque. Although these fibrous alterations have been recognized by light microscopists for almost a century, detailed biochemical and morphological analysis of the lesions has been undertaken only recently. Because the intraneuronal deposits in the NFT and the plaque neurites and the extraneuronal amyloid cores of the plaques have a filamentous ultrastructure, the neuronal cytoskeleton has played a prominent role in most pathogenetic hypotheses.The approach of our laboratory toward elucidating the origin of plaques and tangles in AD has been two-fold: the use of analytical protein chemistry to purify and then characterize the pathological fibers comprising the tangles and plaques, and the use of certain monoclonal antibodies to neuronal cytoskeletal proteins that, despite high specificity, cross-react with NFT and thus implicate epitopes of these proteins as constituents of the tangles.

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