Production of monoclonal antibodies and development of an ELISA for solamargine

被引:70
作者
Ishiyama, M
Shoyama, Y
Murakami, H
Shinohara, H
机构
[1] KYUSHU UNIV,FAC PHARMACEUT SCI,DEPT PHARMACOGNOSY,HIGASHI KU,FUKUOKA 812,JAPAN
[2] KYUSHU UNIV,FAC AGR,GRAD SCH GENET RESOURCES TECHNOL,HIGASHI KU,FUKUOKA 812,JAPAN
关键词
steroidal alkaloid; Solanum khasianum; solamargine; ELISA; monoclonal antibody; qualitative analysis; mass spectrometry;
D O I
10.1007/BF00767762
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
The ratio of hapten and bovine serum albumin in an antigen conjugate was determined by matrix-assisted laser desorption/ionization mass spectrometry. A hybridoma secreting monoclonal antibodies against solamargine was produced by fusing splenocytes immunized with a solamargine-bovine serum albumin conjugate with HAT-sensitive mouse myeloma cell line, P3-X63-Ag8-653. Extensive cross-reaction of anti-solamargine antibodies against solasonine appeared. Aglycone of solamargine, solasodine cross-reacted with anti-solamargine antibodies resulting in a 43.8% cross-reaction. Insignificant cross-reaction appeared with tomatine (2.06%). The full measuring range of the assay extends from 57.5 pmol ml(-1) to 11.5 nmol ml(-1) of solamargine.
引用
收藏
页码:153 / 158
页数:6
相关论文
共 16 条
[1]  
Galfre G, 1981, Methods Enzymol, V73, P3
[2]   ANTIBODY-PRODUCTION BY HYBRIDOMAS [J].
GODING, JW .
JOURNAL OF IMMUNOLOGICAL METHODS, 1980, 39 :285-308
[3]   DETERMINATION OF TETRAHYDROCANNABINOLIC ACID-CARRIER PROTEIN CONJUGATE BY MATRIX-ASSISTED LASER-DESORPTION IONIZATION MASS-SPECTROMETRY AND ANTIBODY-FORMATION [J].
GOTO, Y ;
SHIMA, Y ;
MORIMOTO, S ;
SHOYAMA, Y ;
MURAKAMI, H ;
KUSAI, A ;
NOJIMA, K .
ORGANIC MASS SPECTROMETRY, 1994, 29 (11) :668-671
[4]  
HATANO K, 1989, PLANTA MED, V56, P131
[5]  
LANGONE JJ, 1982, J IMMUNOL METHODS, V51, P33
[6]   STEROIDAL ALKALOIDS FROM SOLANUM-KHASIANUM - APPLICATION OF C-13 NMR-SPECTROSCOPY TO THEIR STRUCTURAL ELUCIDATION [J].
MAHATO, SB ;
SAHU, NP ;
GANGULY, AN ;
KASAI, R ;
TANAKA, O .
PHYTOCHEMISTRY, 1980, 19 (09) :2017-2020
[7]  
MATSUMOTO M, 1989, PLANT CELL REP, V7, P636, DOI 10.1007/BF00272048
[8]   GROWTH OF HYBRIDOMA CELLS IN SERUM-FREE MEDIUM - ETHANOLAMINE IS AN ESSENTIAL COMPONENT [J].
MURAKAMI, H ;
MASUI, H ;
SATO, GH ;
SUEOKA, N ;
CHOW, TP ;
KANOSUEOKA, T .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA-BIOLOGICAL SCIENCES, 1982, 79 (04) :1158-1162
[9]  
SHOYAMA Y, 1993, BIOL PHARM BULL, V16, P1051
[10]   DIRECT DETERMINATION OF FORSKOLIN BOVINE SERUM-ALBUMIN CONJUGATE BY MATRIX-ASSISTED LASER-DESORPTION IONIZATION MASS-SPECTROMETRY [J].
SHOYAMA, Y ;
SAKATA, R ;
ISOBE, R ;
MURAKAMI, H .
ORGANIC MASS SPECTROMETRY, 1993, 28 (09) :987-988