A monoclonal antibody to cyclomaltoheptaose (beta-cyclodextrin): Characterization and use for immunoassay of beta-cyclodextrin and its derivatives.
Cyclomaltoheptaose (beta-cyclodextrin, beta-CD) is a promising compound for application in various industrial fields because of its ability to entrap various compounds into its hydrophobic cavity. A monoclonal antibody (A7) to beta-CD was generated by using a conjugate of glucosaminylmaltosyl-beta-CD and bovine serum albumin as an antigen. The A7 monoclonal antibody was IgM/kappa and reacted with beta-CD with high specificity. The epitope recognized by the A7 monoclonal antibody seemed to be located on the secondary hydroxyl groups of the rim side of the beta-CD molecule. The dissociation constant of the complex of beta-CD and the immobilized A7 monoclonal antibody was determined to be 1.2 x 10(-4) M. A competitive ELISA using the A7 monoclonal antibody enabled determination of beta-CD and its derivatives with a detection limit of 0.05 muM. This immunoassay was useful to determine beta-CD in biological fluids such as human plasma and urine after appropriate pretreatment of the samples.
Ikura
Koji
K
Department of Applied Biology, Kyoto Institute of Technology, Kyoto, Japan, ikura@ipc.kit.ac.jp.
Fujimoto
Jun-Ichi
J
Kubonishi
Kosuke
K
Natsuka
Shunji
S
Hashimoto
Hiroyuki
H
Ito
Tetsuya
T
Fujita
Koki
K