Antibody conjugation to carboxyl-modified microspheres through N-hydroxysuccinimide chemistry for automated immunoassay applications: A general procedure

• DOI: 10.1371/journal.pone.0218686
• Publication Date: 2019-06-26T17:37:55Z
• Authors: Peter Carl; Inês I. Ramos; Marcela A. Segundo; Rudolf J. Schneider
• Keywords: Microtiter plate; Sample Preparation; Bioanalysis

Immunochemical techniques are the workhorse for sample enrichment and detection of a large variety analytes. In contrast to classical microtiter plate-based assays, microparticles next generation solid support, as they promote automation immunoassays using flow-based techniques. Antibody immobilization is crucial step, these reagents expensive, inefficient coupling can result in low sensitivities. This paper proposes general procedure efficient antibodies onto TentaGel particles, via N-hydroxysuccinimide chemistry. The goal was preparation supports with optimum immunorecognition, while increasing sustainability process. influence buffer composition, activation time, well amount antibody on efficiency investigated, resorting fluorophore-labeled proteins fluorescence imaging. Buffer pH time most important parameters coupling. It demonstrated, that hydrolysis esters occurs at similar rates solution, limiting utilizable Finally, applicability generated material automated affinity extraction demonstrated mesofluidic platform lab-on-valve.