title: Site-directed antibody immobilization techniques for immunosensors.
abstract: Immunosensor sensitivity, regenerability, and stability directly depend on the type of antibodies used for the immunosensor design, quantity of immobilized molecules, remaining activity upon immobilization, and proper orientation on the sensing interface. Although sensor surfaces prepared with antibodies immobilized in a random manner yield satisfactory results, site-directed immobilization of the sensing molecules significantly improves the immunosensor sensitivity, especially when planar supports are employed. This review focuses on the three most conventional site-directed antibody immobilization techniques used in immunosensor design. One strategy of immobilizing antibodies on the sensor surface is via affinity interactions with a pre-formed layer of the Fc binding proteins, e.g., protein A, protein G, Fc region specific antibodies or various recombinant proteins. Another immobilization strategy is based on the use of chemically or genetically engineered antibody fragments that can be attached to the sensor surface covered in gold or self-assembled monolayer via the sulfhydryl groups present in the hinge region. The third most common strategy is antibody immobilization via an oxidized oligosaccharide moiety present in the Fc region of the antibody. The principles, advantages, applications, and arising problems of these most often applied immobilization techniques are reviewed.
authors.affiliation: NanoTechnas-Center of Nanotechnology and Materials Science, Department of Analytical and Environmental Chemistry, Faculty of Chemistry, Vilnius University, Naugarduko Street 24, LT-03225 Vilnius, Lithuania.
authors.forename: Asta
authors.initials: A
authors.lastname: Makaraviciute
authors.forename: Almira
authors.initials: A
authors.lastname: Ramanaviciene
mesh_terms: D000818:Animals; D055910:Antibodies, Immobilized / Q000737:chemistry*; D015374:Biosensing Techniques / Q000379:methods*; D019204:GTP-Binding Proteins / Q000737:chemistry; D006801:Humans; D007118:Immunoassay / Q000379:methods*; D013205:Staphylococcal Protein A / Q000737:chemistry
chemical_list: D055910:Antibodies, Immobilized; D013205:Staphylococcal Protein A; D019204:GTP-Binding Proteins
keywords: Antibody; Immunosensor; Site-directed immobilization