MilliporeSigma
  • Detection of Infertility-related Neutralizing Antibodies with a Cell-free Microfluidic Method.

Detection of Infertility-related Neutralizing Antibodies with a Cell-free Microfluidic Method.

Scientific reports (2015-11-21)
Klaus Eyer, Katharina Root, Pascal E Verboket, Petra S Dittrich
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.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Titanium(IV) oxide, nanowires, diam. × L ~10 nm × 10 μm
Sigma-Aldrich
Titanium(IV) oxide, nanowires, diam. × L ~100 nm × 10 μm
Sigma-Aldrich
Titanium(IV) oxide, mixture of rutile and anatase, nanopowder, <100 nm particle size (BET), 99.5% trace metals basis
Sigma-Aldrich
Titanium(IV) oxide, rutile, nanopowder, <100 nm particle size, 99.5% trace metals basis
Sigma-Aldrich
Titanium(IV) oxide, rutile, <001>, (single crystal substrate), ≥99.9% trace metals basis, L × W × thickness 10 mm × 10 mm × 0.5 mm
Sigma-Aldrich
Titanium(IV) oxide, contains 1% Mn as dopant, nanopowder, <100 nm particle size (BET), ≥97%
Sigma-Aldrich
Hexamethyldisiloxane, viscosity 0.65 cSt (25 °C)
Sigma-Aldrich
Sulfuryl chloride, 97%
Sigma-Aldrich
Titanium(IV) oxide, rutile, 99.995% trace metals basis
Sigma-Aldrich
Titanium(IV) oxide, rutile, ≥99.98% trace metals basis
Sigma-Aldrich
Titanium(IV) oxide, nanopowder, 21 nm primary particle size (TEM), ≥99.5% trace metals basis
Sigma-Aldrich
Titanium(IV) oxide, mixture of rutile and anatase, nanoparticles, <150 nm particle size (volume distribution, DLS), dispersion, 40 wt. % in H2O, 99.5% trace metals basis
Sigma-Aldrich
Water, sterile-filtered, BioReagent, suitable for cell culture
Sigma-Aldrich
E-Toxate Water, endotoxin, free
Sigma-Aldrich
Dipyridamole, ≥98% (HPLC)
Sigma-Aldrich
Poly(dimethylsiloxane), viscosity 1.0 cSt (25 °C)
Sigma-Aldrich
Water, for embryo transfer, sterile-filtered, BioXtra, suitable for mouse embryo cell culture
Sigma-Aldrich
Titanium(IV) oxide, rutile, powder, <5 μm, ≥99.9% trace metals basis
Sigma-Aldrich
Water, deuterium-depleted, ≤1 ppm (Deuterium oxide)
Sigma-Aldrich
Water, Nuclease-Free Water, for Molecular Biology
Sigma-Aldrich
Water, PCR Reagent
Sigma-Aldrich
N,N-Dimethylformamide, for molecular biology, ≥99%
Sigma-Aldrich
Water, for molecular biology, sterile filtered
Sigma-Aldrich
Water, for cell biology, sterile ultrafiltered
Sigma-Aldrich
Water, ACS reagent
Sigma-Aldrich
N,N-Dimethylformamide, puriss. p.a., ACS reagent, reag. Ph. Eur., ≥99.8% (GC)
Sigma-Aldrich
N,N-Dimethylformamide, biotech. grade, ≥99.9%
Sigma-Aldrich
N,N-Dimethylformamide, ACS reagent, ≥99.8%
Sigma-Aldrich
Water, Deionized
Sigma-Aldrich
N,N-Dimethylformamide, ReagentPlus®, ≥99%