Journal: International Journal of Nanomedicine

Article Title: Development of a Sensitive Quantum Dot-Linked Immunoassay for the Multiplex Detection of Biochemical Markers in a Microvolumeric Format

doi: 10.2147/ijn.s477118

Figure Lengend Snippet: Scheme 1 Scheme of the classical QLISA detection method. COMP (cartilage oligomeric matrix protein) and hGH (human growth hormone) proteins are attached to the bottom of the plate, incubated with biotinylated detection antibodies, and then conjugated with streptavidin-coated QDs, followed by the direct registration of fluorescence signal from the bottom of the plate.

Article Snippet: Prior to the experiment, the gold surface of the SPR sensor disc (SD AU, XanTec bioanalytics GmbH, Münster, Germany) was cleaned, modified with a self-assembled monolayer of 11-mercaptoundecanoic acid (Sigma-Aldrich, Steinheim, Germany) and capture antibodies against COMP (monoclonal mouse IgG1 clone 16F12, BioVendor, Brno, Czech Republic; Cat. No. RD182080100F1-01) were immobilized.

Techniques: Incubation, Fluorescence

Journal: International Journal of Nanomedicine

Article Title: Development of a Sensitive Quantum Dot-Linked Immunoassay for the Multiplex Detection of Biochemical Markers in a Microvolumeric Format

doi: 10.2147/ijn.s477118

Figure Lengend Snippet: Scheme 2 Singleplex detection of an analyte in microvolume format. Formation of COMP (cartilage oligomeric matrix protein), or hGH (human growth hormone) complexes with biotin-labeled anti-COMP or anti-hGH antibodies, respectively, and streptavidin-coated QDs of different spectra. Immune-complex disassembling solution dissociates the complexes through an analyte-biotinylated antibody connection. After dissociation, microdrops are transferred to microvolume reusable glass slide-based spectrophotometric scan.

Article Snippet: Prior to the experiment, the gold surface of the SPR sensor disc (SD AU, XanTec bioanalytics GmbH, Münster, Germany) was cleaned, modified with a self-assembled monolayer of 11-mercaptoundecanoic acid (Sigma-Aldrich, Steinheim, Germany) and capture antibodies against COMP (monoclonal mouse IgG1 clone 16F12, BioVendor, Brno, Czech Republic; Cat. No. RD182080100F1-01) were immobilized.

Techniques: Labeling

Journal: International Journal of Nanomedicine

Article Title: Development of a Sensitive Quantum Dot-Linked Immunoassay for the Multiplex Detection of Biochemical Markers in a Microvolumeric Format

doi: 10.2147/ijn.s477118

Figure Lengend Snippet: Figure 3 Optimization of COMP analyte detection on the microslide. Non-diluted and diluted 1:1 (v/v) with PBS 10 mM NaOH and 0.5% SDS buffer on a glass microslide. At concentrations 50 nM, 25 nM, and 5 nM COMP was analyzed using 17C10 anti-COMP-biotinylated Ab and streptavidin-coated QD655. MFI – median fluorescence intensity; Blank – sample without COMP protein coating, followed by all of the other protocol steps. P-values indicate statistical significance (***P ≤ 0.001). Error bars represent the standard deviation of the average value (N=3).

Article Snippet: Prior to the experiment, the gold surface of the SPR sensor disc (SD AU, XanTec bioanalytics GmbH, Münster, Germany) was cleaned, modified with a self-assembled monolayer of 11-mercaptoundecanoic acid (Sigma-Aldrich, Steinheim, Germany) and capture antibodies against COMP (monoclonal mouse IgG1 clone 16F12, BioVendor, Brno, Czech Republic; Cat. No. RD182080100F1-01) were immobilized.

Techniques: Fluorescence, Standard Deviation

Journal: International Journal of Nanomedicine

Article Title: Development of a Sensitive Quantum Dot-Linked Immunoassay for the Multiplex Detection of Biochemical Markers in a Microvolumeric Format

doi: 10.2147/ijn.s477118

Figure Lengend Snippet: Figure 4 Different COMP concentration (50 nM to 3.125 nM) detection in 96-well plate vs microvolumetric format. 17C10 anti-COMP-biotinylated Ab and streptavidin- coated QD655 were used. Purple bars indicate the signal detected from the bottom of the plate, excitation 400 nm, emission 655 nm, 80 μL volume. Blue bars indicate the signal detected on a microvolume glass slide (2 µL/sample), excitation 400 nm, emission 655 nm. Blank – every protocol step followed, excluding coating the bottom of the well with COMP (non-specific signal). P-values indicate statistical significance (****P < 0.0001). Error bars represent the standard deviation of the average value (96-well plate N=2; microvolumes N=3).

Article Snippet: Prior to the experiment, the gold surface of the SPR sensor disc (SD AU, XanTec bioanalytics GmbH, Münster, Germany) was cleaned, modified with a self-assembled monolayer of 11-mercaptoundecanoic acid (Sigma-Aldrich, Steinheim, Germany) and capture antibodies against COMP (monoclonal mouse IgG1 clone 16F12, BioVendor, Brno, Czech Republic; Cat. No. RD182080100F1-01) were immobilized.

Techniques: Concentration Assay, Standard Deviation