Use of biosensor as a tool for double antibody sandwich immunoassay establishment:

Abstract

Background: For biomarker detection, double antibody sandwich platform is the most sensitive and specific immunoassay. Development of double antibody sandwich platform requires the step of appropriate antibody pair selection. However, identification of matched antibody pairs is difficult, laborious and time-consuming. Objectives: To illustrate the application of biosensor for selecting the suitable matched pairs of antibody for double antibody sandwich platform. Materials and methods: Double antibody sandwich immunoassay for Ag85B protein detection was selected as a model study. Biolayer interferometry biosensor was used to search matched pairs of antibody for Ag85B protein detection. To prove the obtained information, the double antibody sandwich ELISAs were employed. Results: Biosensor results revealed that the best-matched antibody pair might be mAb clone AM85B-8B as capture antibody and mAb clone AM85B-5B or AM85B-9B as detecting antibody. The results obtained from biosensor system and ELISA were similar. Conclusion: Biosensor can be used as a tool for effectively screening and selecting the best pairs of antibodies for any immunoassays. Biosensor method is very useful for reducing processes and time for double antibody immunoassay development.

Keywords

• double antibody sandwich immunoassay
• immunoassay development
• biosensor