Oligosaccharide-assisted direct immunosensing of small molecules

Anal Chem. 2010 Jun 1;82(11):4333-6. doi: 10.1021/ac100865p.

Abstract

"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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Biosensing Techniques / methods*
  • Carbohydrate Conformation
  • Cattle
  • Haptens / analysis
  • Haptens / chemistry
  • Haptens / immunology
  • High-Throughput Screening Assays
  • Immunoassay / methods*
  • Models, Molecular
  • Oligosaccharides / metabolism*
  • Serum Albumin, Bovine / metabolism
  • Single-Chain Antibodies / immunology
  • Single-Chain Antibodies / metabolism
  • beta-Cyclodextrins / chemistry
  • beta-Cyclodextrins / metabolism

Substances

  • Haptens
  • Oligosaccharides
  • Single-Chain Antibodies
  • beta-Cyclodextrins
  • Serum Albumin, Bovine