Development and Application of an Enzyme Immunoassay for Karasurin A, an Effective Protein Component of Trichosanthes kirilowii MAX. var. japonicum KITAM

Development and Application of an Enzyme Immunoassay for Karasurin A, an Effective Protein Component of Trichosanthes kirilowii MAX. var. japonicum KITAM

A method was developed for specific estimation of the content of a non-enzymatic protein, karasurin A, in fractions taken during the extraction and purification processes from a natural source. Anti-karokon serum was elicited in rabbits immunized with fragments of karokon, a dried root tuber of Tric...

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Bibliographic Details
Published in:Chemical & pharmaceutical bulletin Vol. 42; no. 3; pp. 599 - 603
Main Authors: KITAGAWA, Tsunehiro, YOSIDA, Masayo, HU, Jiango, SAKAI, Atsuko, BAY, Gang, FUJIWARA, Kunio, OGIHARA, Yukio, TAKEDA, Tadahiro
Format: Journal Article
Language:English
Published: Tokyo The Pharmaceutical Society of Japan 1994
Maruzen
Japan Science and Technology Agency
Subjects:
Animals
anti-karokon serum
Biological and medical sciences
Chinese crude drug
Drugs, Chinese Herbal - analysis
General pharmacology
Immunoenzyme Techniques
karasurin A
Medical sciences
N-Glycosyl Hydrolases
Pharmacognosy. Homeopathy. Health food
Pharmacology. Drug treatments
Plant Proteins - analysis
Plant Proteins - immunology
Rabbits - immunology
selected antibody enzyme immunoassay
Trichosanthes kirilowii
Tropical medicine
Asia
China
Root
Pharmacognosy
Homogenate
Cucurbitaceae
Enzyme immunoassay
Medicinal plant
Proteins
Folk medicine
Dicotyledones
Angiospermae
Plant origin
Spermatophyta
Quantitative analysis
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Summary:A method was developed for specific estimation of the content of a non-enzymatic protein, karasurin A, in fractions taken during the extraction and purification processes from a natural source. Anti-karokon serum was elicited in rabbits immunized with fragments of karokon, a dried root tuber of Trichosanthes kirilowii MAX. var. japonicum KITAM. Rabbit antibody specific for karasurin A was identified in anti-karokon serum by the Western blotting method. After separation by SDS-PAGE, protein bands of purified karasurin A and extracted proteins from a medicinal herb which is a karasurin A source were reacted with anti-karokon serum followed by treatment with horseradish peroxidase (HRP)-labeled Fab' of goat anti-rabbit IgG, and then bound HRP-labeled second antibody on protein bands was developed to brown by reaction with a substrate solution of the used enzyme. A novel selected antibody enzyme immunoassay (SAEIA) for karasurin A was developed using selective binding of anti-karasurin A antibody in anti-karokon serum to solid phase karasurin A and HRP-labeled Fab' of the second antibody as the tracer. Specific estimation of the content of karasurin A in several fractions taken during the isolation and purification processes of the protein were possible using the SAEIA method.
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content type line 23
ISSN:0009-2363
1347-5223
DOI:10.1248/cpb.42.599