title: Detection of Infertility-related Neutralizing Antibodies with a Cell-free Microfluidic Method.
abstract: The unwanted emergence of neutralizing antibodies (nAbs) against an endogenous or a therapeutic protein can result in deficiency diseases or therapy failure. Here, we developed a cell-free microfluidic method for the sensitive detection and quantification of nAbs in human serum that are associated with infertility. We used cell-derived vesicles containing the luteinizing hormone (LH)/choriogonadotropin receptor (LHHCGR) to detect nAbs against LH. The method exploits the entire cellular signal amplification mechanism, and facilitates the detection of as little as 0.44 nM of LH-nAb (Kd 1.5 nM) in human serum matrix within only 15 minutes. In addition, dose-response curves can be generated in less than 2 hours to evaluate the nAB concentration and dissociation constant. The developed system is devoid of problems associated with cell-based assays and we believe that this simple effect-directed analysis can be used in clinical environments, and is adaptable to other hormones or cytokines and their respective nAbs.
authors.affiliation: Department of Chemistry and Applied Biosciences, ETH Zurich Switzerland.
authors.forename: Klaus
authors.initials: K
authors.lastname: Eyer
authors.affiliation: Department of Chemistry and Applied Biosciences, ETH Zurich Switzerland.
authors.forename: Katharina
authors.initials: K
authors.lastname: Root
authors.affiliation: Department of Chemistry and Applied Biosciences, ETH Zurich Switzerland.|Department of Biosystems Science and Engineering, ETH Zurich Switzerland.
authors.forename: Pascal E
authors.initials: PE
authors.lastname: Verboket
authors.affiliation: Department of Chemistry and Applied Biosciences, ETH Zurich Switzerland.|Department of Biosystems Science and Engineering, ETH Zurich Switzerland.
authors.forename: Petra S
authors.initials: PS
authors.lastname: Dittrich
mesh_terms: D057134:Antibodies, Neutralizing / Q000097:blood / Q000276:immunology*; D055252:Cell-Derived Microparticles / Q000276:immunology*; D057809:HEK293 Cells; D006801:Humans; D007246:Infertility / Q000097:blood / Q000276:immunology*; D008163:Luminescent Measurements / Q000379:methods; D007986:Luteinizing Hormone / Q000276:immunology; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation / Q000379:methods*; D011974:Receptors, LH / Q000276:immunology; D015203:Reproducibility of Results; D020349:Surface Plasmon Resonance
chemical_list: D057134:Antibodies, Neutralizing; D011974:Receptors, LH; D007986:Luteinizing Hormone