Production of monoclonal antibodies against a major purgative component, sennoside B, their characterization and use in ELISA.
For immunization, sennoside B was conjugated with bovine serum albumin. The hapten density in the antigen conjugate was determined to be 3 mol mol(-1) protein by matrix-assisted laser desorption-ionization TOF mass spectrometry. A hybridoma secreting monoclonal antibody against sennoside B was produced by fusing splenocytes from mouse immunized with the sennoside B conjugate and mouse myeloma cells. Weak cross-reactivities occurred with sennoside A which is a stereochemical isomer, and a monomer of sennoside B, rhein, but no cross-reactivity was observed with other related anthraquinones and phenolics. The range of the assay extended from 0.5 ng ml(-1) to 15 ng ml(-1) of sennoside B, and good correlation between ELISA and HPLC methods was obtained when crude extracts of rhubarb were analyzed.
Morinaga
O
O
Department of Pharmacognosy, Graduate School of Pharmaceutical Sciences, Kyushu University, Fukuoka, Japan.
Nakajima
S
S
Tanaka
H
H
Shoyama
Y
Y
D000880:Anthraquinones / Q000032:analysis / Q000276:immunology*; D000911:Antibodies, Monoclonal / Q000302:isolation & purification*; D002400:Cathartics / Q000032:analysis*; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D012676:Senna Extract; D000081226:Sennosides
D000880:Anthraquinones; D000911:Antibodies, Monoclonal; D002400:Cathartics; D000081226:Sennosides; D012676:Senna Extract