Protein A used in DELFIA for the determination of specific antibodies

Immunol Lett. 1993 Mar;35(3):285-9. doi: 10.1016/0165-2478(93)90196-9.

Abstract

The conditions of protein A labelling with Eu chelates were studied. The conjugates obtained were compared with those from horseradish peroxidase used conventionally in immunochemical practice. Protein A-Eu conjugates were obtained by a method applied previously for antibody labelling with indium and europium chelates using the bicyclic dianhydride of diethylenetriaminepenta-acetic acid (DADTPA) with some modifications. The Eu-labeled protein A ensured a sensitivity of the IgG determination at the level of 2 ng/ml and a dynamic range of the determination from 3 to 1000 ng/ml, which significantly exceed analogous values for the protein A-peroxidase conjugates. The Eu-labeled protein A was used for the determination of antibodies to Francisella tularensis in the sera of humans vaccinated against tularemia. The assay values exceeded by 10-40-fold the results of an ELISA in sensitivity. It was deduced that the Eu-labeled protein A can be effectively used for the determination of antibodies specific to a tularemia causative agent. In particular, this compound can be useful for the determination of specific antibodies in low immune sera.

Publication types

  • Comparative Study

MeSH terms

  • Antibodies, Bacterial / analysis
  • Enzyme-Linked Immunosorbent Assay
  • Europium
  • Fluorescent Antibody Technique*
  • Francisella tularensis / immunology
  • Humans
  • Recombinant Proteins
  • Staphylococcal Protein A*
  • Tularemia / immunology

Substances

  • Antibodies, Bacterial
  • Recombinant Proteins
  • Staphylococcal Protein A
  • Europium