Development of competitive ELISA for the detection of bovine serum albumin using single-chain variable fragments

Anal Biochem. 2017 May 15:525:89-91. doi: 10.1016/j.ab.2017.03.003. Epub 2017 Mar 6.

Abstract

Soluble anti-bovine serum albumin (BSA) single-chain variable fragments (scFvs) were expressed in E. Coli. HB2151. The antigen-binding equilibrium dissociation constant of the scFvs was determined to be 2.9 × 10-8 M by surface plasmon resonance analysis. A competitive ELISA for the detection of BSA was developed using the antibody fragment above. The limits of detection (I10) and I50 were 0.002 and 0.74 μg/ml respectively, with a recovery between 87.8 and 119.2% in spiked milk samples. The assay has the potential to be used to detect concentration of BSA in milk or other matrix instead of the ELISA based on traditional antibodies.

Keywords: Bovine serum albumin; ELISA; Single-chain variable fragments.

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Antibodies, Monoclonal / immunology
  • Cattle
  • Enzyme-Linked Immunosorbent Assay / methods*
  • Escherichia coli / metabolism*
  • Milk / chemistry*
  • Milk Proteins / analysis*
  • Serum Albumin, Bovine / analysis*
  • Serum Albumin, Bovine / immunology
  • Single-Chain Antibodies / immunology*
  • Surface Plasmon Resonance

Substances

  • Antibodies, Monoclonal
  • Milk Proteins
  • Single-Chain Antibodies
  • Serum Albumin, Bovine