Concise synthesis of ciguatoxin ABC-ring fragments and surface plasmon resonance study of the interaction of their BSA conjugates with monoclonal antibodies.
Monoclonal antibodies (mAbs), 4H2 and 6H7, were prepared previously using a protein conjugate of a 1:1 epimeric mixture of the synthetic ABC-ring fragments of ciguatoxin (CTX), 3 and 4. Here, the interactions of these mAbs with the fragments of CTX and CTX3C, 3 and 5, were investigated by surface plasmon resonance (SPR) spectroscopy in an attempt to clarify an antigenic determinant. Compared with the previous synthesis, the fragment 3 possessing the 2S configuration was synthesized from tri-O-acetyl-D-glucal much more effectively. The mAb 4H2 was already known to show a dose-dependent binding to the bovine serum albumin (BSA) conjugate of 3, but not to that of 5. The present SPR study of 4H2 demonstrates that the A-ring side chain of 3 plays a decisive role as an epitope. Therefore, SPR can effectively replace the ELISA method for the analysis of mAbs.
Nagumo
Y
Y
Department of Chemistry, Graduate School of Science, Tohoku University, and CREST, Japan Science and Technology Corporation (JST), 980-8578, Sendai, Japan.
Oguri
H
H
Shindo
Y
Y
Sasaki
S
S
Oishi
T
T
Hirama
M
M
Tomioka
Y
Y
Mizugaki
M
M
Tsumuraya
T
T
D000911:Antibodies, Monoclonal / Q000032:analysis / Q000737:chemistry / Q000378:metabolism*; D001666:Binding Sites, Antibody / Q000502:physiology; D002922:Ciguatoxins / Q000138:chemical synthesis / Q000378:metabolism*; D003847:Deoxyglucose / Q000031:analogs & derivatives / Q000138:chemical synthesis; D004305:Dose-Response Relationship, Drug; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes / Q000737:chemistry / Q000378:metabolism*; D008968:Molecular Conformation; D012710:Serum Albumin, Bovine / Q000737:chemistry / Q000378:metabolism*; D020349:Surface Plasmon Resonance
D000911:Antibodies, Monoclonal; D000939:Epitopes; D002922:Ciguatoxins; C001289:d-glucal; D012710:Serum Albumin, Bovine; D003847:Deoxyglucose