Protein A used in DELFIA for the determination of specific antibodies.

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.