title: Single chain fragment variable recombinant antibody as a template for Fc sensors.
abstract: A label free immunosensor for detection of Fc receptors expressed on cell surface was developed and characterized using a Quartz Crystal Microbalance (QCM) transducer. Taking advantage of the characteristics of single chain fragment variable (scFv) recombinant antibody and the multivalency of an antibody, the engineered recombinant scFv was immobilized onto preformed functionalized self-assembled monolayers (SAMs) template surface. The monomeric scFv can bind with the CH1 region of any rabbit IgG to form a highly oriented IgG layer with its Fc portion pointing toward a solution phase. This results in a highly oriented Fc sensor that can be used to study the thermodynamics and kinetics of binding between the Fc portion of immunoglobulin and the cell surface Fc receptor (FcR), an important area of the immune system. The Fc sensor was used to study the binding between Staphylococcus aureus and the Fc receptor on macrophage. Parallel characterization of cell surface Fc receptors in the same samples by ELISA was also performed.
authors.affiliation: Department of Biochemistry, School of Medicine, Vanderbilt University, Nashville, Tennessee 37232, USA.
authors.forename: Heping
authors.initials: H
authors.lastname: Yan
authors.forename: Zhihong
authors.initials: Z
authors.lastname: Shen
authors.forename: Ray
authors.initials: R
authors.lastname: Mernaugh
authors.forename: Xiangqun
authors.initials: X
authors.lastname: Zeng
mesh_terms: D000818:Animals; D055910:Antibodies, Immobilized / Q000737:chemistry; D015374:Biosensing Techniques / Q000379:methods*; D007141:Immunoglobulin Fc Fragments / Q000737:chemistry*; D007074:Immunoglobulin G / Q000737:chemistry / Q000276:immunology; D007700:Kinetics; D011485:Protein Binding; D058525:Quartz Crystal Microbalance Techniques / Q000379:methods; D011817:Rabbits; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D057127:Single-Chain Antibodies / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D013211:Staphylococcus aureus / Q000302:isolation & purification; D013816:Thermodynamics
chemical_list: D055910:Antibodies, Immobilized; D007141:Immunoglobulin Fc Fragments; D007074:Immunoglobulin G; D011994:Recombinant Proteins; D057127:Single-Chain Antibodies