Журнал аналитических и биоаналитических методов

Абстрактный

A Rapid ELISA Method to Improve the Automated Test Throughput

Daniele Casini, Paola Fontani, Paolo Ruggiero, Enrico Balducci and Duccio Berti

In ELISA, ligand is commonly adsorbed to the plastic surface through non-covalent bonds between the hydrophobic regions of the ligand and the plastic surface. Thus, all the reactions occur in a heterogeneous phase, with some reactants in solution, and some immobilized. As a result, the diffusion constant of immobilized reagents is zero and the overall reaction speed is low. With the proposed new approach the first step occurs in a homogeneous phase, aimed at speeding up ELISA procedure especially to easily adapt it to robotic systems. Conventional tests usually may take up to 5 hours. Our “rapid ELISA” approach considerably reduces this time to less than 30 minutes allowing the method to be more suitable for automation. The rapid ELISA has been set up to analyze samples coming from animal studies for vaccine development purposes. In particular, it was applied to quantitate antibodies specific for the Outer Membrane Vesicle of Neisseria meningitidis group B and Influenza virus antigens (H1N1; H3N2; B). Due to its high flexibility, this rapid ELISA can be used to detect a wide range of antibodies raised against a wide range of antigens.