跳转至内容

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

摘要

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.

材料

货号

品牌

产品描述

Sigma-Aldrich

水, Nuclease-Free Water, for Molecular Biology

Sigma-Aldrich

N,N-二甲基甲酰胺, ACS reagent, ≥99.8%

Sigma-Aldrich

水, sterile-filtered, BioReagent, suitable for cell culture

Sigma-Aldrich

3-异丁基-1-甲基黄嘌呤, ≥99% (HPLC), powder

Sigma-Aldrich

水, Deionized

Sigma-Aldrich

N,N-二甲基甲酰胺, puriss. p.a., ACS reagent, reag. Ph. Eur., ≥99.8% (GC)

Sigma-Aldrich

3-异丁基-1-甲基黄嘌呤, ≥99%, BioUltra

Sigma-Aldrich

水, for embryo transfer, sterile-filtered, BioXtra, suitable for mouse embryo cell culture

Sigma-Aldrich

N,N-二甲基甲酰胺, anhydrous, 99.8%

Sigma-Aldrich

N,N-二甲基甲酰胺, ReagentPlus^®, ≥99%

Sigma-Aldrich

N,N-二甲基甲酰胺, for molecular biology, ≥99%

Sigma-Aldrich

水, for molecular biology, sterile filtered

Sigma-Aldrich

氧化钛（IV）, nanopowder, 21 nm primary particle size (TEM), ≥99.5% trace metals basis

Sigma-Aldrich

腺苷, ≥99%

Sigma-Aldrich

叠氮化钠, BioUltra, ≥99.5% (T)

Sigma-Aldrich

叠氮化钠, ReagentPlus^®, ≥99.5%

Sigma-Aldrich

氧化钛 (IV)，锐钛矿, nanopowder, <25 nm particle size, 99.7% trace metals basis

Sigma-Aldrich

N,N-二甲基甲酰胺, biotech. grade, ≥99.9%

Sigma-Aldrich

氧化钛（IV）, puriss., meets analytical specification of Ph. Eur., BP, USP, 99-100.5%

Sigma-Aldrich

叠氮化钠, purum p.a., ≥99.0% (T)

Sigma-Aldrich

氧化钛 (IV)，锐钛矿, powder, 99.8% trace metals basis

Sigma-Aldrich

氧化钛（IV）, ReagentPlus^®, ≥99%

Sigma-Aldrich

水, BioPerformance Certified

Sigma-Aldrich

钛 (IV) 氧化物，金红石型, powder, <5 μm, ≥99.9% trace metals basis

Sigma-Aldrich

氧化钛 (IV)，锐钛矿, powder, −325 mesh, ≥99% trace metals basis

Sigma-Aldrich

钛 (IV) 氧化物，金红石型, nanopowder, <100 nm particle size, 99.5% trace metals basis

Sigma-Aldrich

腺苷-3′,5′-环单磷酸, ≥98.5% (HPLC), powder

Sigma-Aldrich

水, ACS reagent

Sigma-Aldrich

腺苷, suitable for cell culture, BioReagent

Sigma-Aldrich

氧化钛(IV)，金红石和锐钛矿混合物, nanopowder, <100 nm particle size (BET), 99.5% trace metals basis