Design of 3D structure membrane for the increased sensitivity in Enzyme Linked Immunosorbent Assay (mELISA)

Abstract

The Enzyme Linked Immunosorbent Assay (ELISA) technique has been widely used for the identification and quantification of biochemical markers. The typical ELISA requires a number of washing steps to eliminate the unbound proteins which sometimes cause the desorption of protein due to their weak bonding between protein and well plate. In this study, we have developed a meshed type of plastic membrane in order to increase the reliable binding efficiency between proteins and the membrane surface, and to provide easy steps of washing. The use of our developed solid membrane has significantly increased the binding capacity of the biomolecules because this membrane ELISA (mELISA) provides 3D binding surfaces which increases the surface area when compared to the conventional 2D surface well plate. The columns were pretreated to form a self-assembled layer (SAM) on the surface for the stable conjugation of a target antibody. The SAM-coated membranes could be stored for one month without any further deterioration of stability. The measured optical density (O.D.) shows a 1.2-fold increase in IgG antigen (25 μg/mL) from the plastic membrane as compared with the conventional ELISA method. The concentrations of thyroid stimulating hormone were also monitored using the mELISA method and it shows good linearity against the concentrations. © 2019 by the authors.