Oligosaccharide-assisted direct immunosensing of small molecules.
"Sandwich-type" noncompetitive (immunometric) assays allow for high-sensitivity high-throughput macromolecule sensing and determination but cannot be used on small molecules (haptens). Here, we isolated single-chain Fv fragments from a phage-display library, which bound to complexes of particular haptens (vitamin D and A derivatives) with immobilized beta-cyclodextrin or beta-maltosyl residues, and formed ternary complexes. These scFvs enabled novel "semisandwich-type" immunometric assays of haptens with nanomole-range sensitivities.
Kobayashi
Norihiro
N
Kobe Pharmaceutical University, 4-19-1, Motoyama-Kitamachi, Higashinada-ku, Kobe 658-8558, Japan. no-kobay@kobepharma-u.ac.jp
Oyama
Hiroyuki
H
Suzuki
Iwao
I
Kato
Yoshinori
Y
Umemura
Takeo
T
Goto
Junichi
J
D000818:Animals; D015374:Biosensing Techniques / Q000379:methods*; D002236:Carbohydrate Conformation; D002417:Cattle; D006241:Haptens / Q000032:analysis / Q000737:chemistry / Q000276:immunology; D057166:High-Throughput Screening Assays; D007118:Immunoassay / Q000379:methods*; D008958:Models, Molecular; D009844:Oligosaccharides / Q000378:metabolism*; D012710:Serum Albumin, Bovine / Q000378:metabolism; D057127:Single-Chain Antibodies / Q000276:immunology / Q000378:metabolism; D047392:beta-Cyclodextrins / Q000737:chemistry / Q000378:metabolism
D006241:Haptens; D009844:Oligosaccharides; D057127:Single-Chain Antibodies; D047392:beta-Cyclodextrins; D012710:Serum Albumin, Bovine