Electrophoretic Screen and Functionalized Microspheres as Antigen Assays

Immunoassays are used for medical diagnosis of infectious diseases. People in developing countries usually do not have access to this type of technologies, which makes infectious diseases the main cause of death in these countries. Immunoassay techniques assist the medical field with diagnosing, prognosis and monitoring of analytes present in the human body. This project will develop a portable antigen detection assay built using an electrophoretic screen with functionalized microspheres. A portable electrophoretic immunoassay will permit the reduction of lab materials and the independence of lab installations making it more accessible. This project involves two main steps: The screen modification by silanation, and the antibodies’ attachment to the microspheres. The silanation process was adapted from Bioconjugate Techniques [1]. Finally, Anti-Chorionic Gonadotropin (HCG) α and β strands antibodies with a concentration of 40 mg/ml are tagged with a fluorescent tag using Atto 495. The remaining particles are separated with a HiTrap Sephadex G-25 gel column. Using 2-MEA antibodies are cleaved at disulfide bridges at the constant region to create sulfhydryls for coupling. Antibodies are then covalently attached to the functionalized microspheres and in the silica screen. Attachment is verified with a con focal microscope. Preliminary results have proven that targeted antibody attachment to silica screen is possible; these results will be presented