human muc1 protein  (ACROBiosystems)

Journal: Advanced Science

Article Title: Analysis of Clinical Samples of Pancreatic Cyst's Lesions with A Multi‐Analyte Bioelectronic Simot Array Benchmarked Against Ultrasensitive Chemiluminescent Immunoassay

doi: 10.1002/advs.202308141

Figure Lengend Snippet: a) Image of the portable SiMoT ELISA‐like 96‐sensors bioelectronic array, developed within the SiMBiT project framework. b) Overhead perspective of the 96 Electrolyte‐Gated Organic field effect transistors (EGOFETs) defined on a PEN substrate with the bottomless ELISA plate glued on top. c) Overhead perspective of the 3D‐sensing gate used as SiMoT array lid. d) Enlarged view of a designated region for the assay of one patient's plasma or cyst's fluid sample, encompassing 16 sensing gates. The color indicators represent biofunctionalized gates tested in triplicate, incorporating specific antibodies targeting MUC1 (cyan) and CD55 (green), or probes designed for the binding of mutated KRAS (red). Furthermore, each patient's sample is subjected to 7 gates coated with BSA (black) for negative‐control experiments.

Article Snippet: Monoclonal antibodies for Mucin 1 and the recombinant protein of human MUC1 were supplied by OriGene, while CD‐55 monoclonal antibodies and the human CD55 proteins were contributed by Abnova.

Techniques: Enzyme-linked Immunosorbent Assay, Binding Assay, Negative Control

Journal: Advanced Science

Article Title: Analysis of Clinical Samples of Pancreatic Cyst's Lesions with A Multi‐Analyte Bioelectronic Simot Array Benchmarked Against Ultrasensitive Chemiluminescent Immunoassay

doi: 10.1002/advs.202308141

Figure Lengend Snippet: Multivariate data processing of the SiMoT and SIMOA assays a) Score‐Plot illustrating the scores of individual samples assessed using the SiMoT array for principal components PC1 and PC2. Red‐labelled samples correspond to high‐grade mucinous cysts, orange labels represent low‐grade mucinous cysts, and blue labels indicate non‐mucinous samples. b) Loading plot displaying the loadings of each initial variable extracted for the SiMoT assay on PC1 and PC2; R I , R D , and R B are marked with cyan, green, and red arrows for MUC1, CD55, and KRAS, respectively. c) Score plot showing the scores of PC1 and PC2 obtained on the SIMOA assay on the 39 patients specimens. d) Loading plot illustrating the loadings of each original variable extracted for the SIMOA assay on PC1 and PC2; R i and R B are indicated with cyan and green arrows for MUC1 and CD55, respectively.

Article Snippet: Monoclonal antibodies for Mucin 1 and the recombinant protein of human MUC1 were supplied by OriGene, while CD‐55 monoclonal antibodies and the human CD55 proteins were contributed by Abnova.

Techniques:

Journal: Advanced Science

Article Title: Analysis of Clinical Samples of Pancreatic Cyst's Lesions with A Multi‐Analyte Bioelectronic Simot Array Benchmarked Against Ultrasensitive Chemiluminescent Immunoassay

doi: 10.1002/advs.202308141

Figure Lengend Snippet: Pictorial representations and Surface Plasmon Resonance (SPR) characterizations of the biofunctionalization protocols involving a–c) anti‐MUC1 and d–f) anti‐CD55 capturing antibodies, while panel g–i) pertains to b‐KRAS probes. The assays for MUC1, CD55, and KRAS standard solutions were conducted, examining a concentration range from 20 nM to 150 nM. All sensograms are presented as the average of two replicates.

Article Snippet: Monoclonal antibodies for Mucin 1 and the recombinant protein of human MUC1 were supplied by OriGene, while CD‐55 monoclonal antibodies and the human CD55 proteins were contributed by Abnova.

Techniques: SPR Assay, Concentration Assay

Journal: Advanced Science

Article Title: Analysis of Clinical Samples of Pancreatic Cyst's Lesions with A Multi‐Analyte Bioelectronic Simot Array Benchmarked Against Ultrasensitive Chemiluminescent Immunoassay

doi: 10.1002/advs.202308141

Figure Lengend Snippet: Surface coverage calculated for the three biofunctionalization procedures with anti‐MUC1, anti‐CD55, and b‐KRAS.

Article Snippet: Monoclonal antibodies for Mucin 1 and the recombinant protein of human MUC1 were supplied by OriGene, while CD‐55 monoclonal antibodies and the human CD55 proteins were contributed by Abnova.

Techniques:

Journal: Advanced Science

Article Title: Analysis of Clinical Samples of Pancreatic Cyst's Lesions with A Multi‐Analyte Bioelectronic Simot Array Benchmarked Against Ultrasensitive Chemiluminescent Immunoassay

doi: 10.1002/advs.202308141

Figure Lengend Snippet: a) Typical sensing signal (red curves) and baseline signal (black curves) observed throughout the cycling process, involving 20 transfer characteristics (I D versus V GS in the 0 V to −0.5 V window at a stable V D = −0.4 V). The baseline is recorded using a 3D sensing gate biofunctionalized with anti‐MUC1 following incubation in PBS reference fluid. The sensing signal is taken using the same 3D sensing gate following incubation in a PBS standard solution of MUC1, containing 12 ± 3 molecules in 0.1 mL. The relative current change R I = (I – I 0 )/ I 0 is plotted against the cycling index for b) the bare gold lateral reference gate, c) the anti‐MUC1 biofunctionalized 3D sensing gate, and d) the BSA‐coated 3D sensing gate used as a negative control experiment. R I values are recorded at V GS and V D of −0.4 V, for each cycling index. Average values for data points were obtained from three replicates across three separate gates, and error bars were calculated as relative standard deviations. The data recorded after incubation in the reference fluid (PBS) is denoted by a black shadow, while measurements following incubation in a PBS standard solution of the target analyte are represented by a red shadow. The dashed‐dotted grey line represents the Limit‐of‐Identification (LOI) threshold.

Article Snippet: Monoclonal antibodies for Mucin 1 and the recombinant protein of human MUC1 were supplied by OriGene, while CD‐55 monoclonal antibodies and the human CD55 proteins were contributed by Abnova.

Techniques: Incubation, Negative Control

Journal: Advanced Science

Article Title: Analysis of Clinical Samples of Pancreatic Cyst's Lesions with A Multi‐Analyte Bioelectronic Simot Array Benchmarked Against Ultrasensitive Chemiluminescent Immunoassay

doi: 10.1002/advs.202308141

Figure Lengend Snippet: List of the patients’ specimens, along with the state‐of‐the‐art diagnosis. Average values of the R I , R D and R B extracted for the SiMoT assay of MUC1, KRAS, and CD55, and of R i and R B extracted for the SIMOA assay of MUC1 and CD55 over three replicates.

Article Snippet: Monoclonal antibodies for Mucin 1 and the recombinant protein of human MUC1 were supplied by OriGene, while CD‐55 monoclonal antibodies and the human CD55 proteins were contributed by Abnova.

Techniques:

Journal: Advanced Science

Article Title: Analysis of Clinical Samples of Pancreatic Cyst's Lesions with A Multi‐Analyte Bioelectronic Simot Array Benchmarked Against Ultrasensitive Chemiluminescent Immunoassay

doi: 10.1002/advs.202308141

Figure Lengend Snippet: a) Pictorial representation of 3D sensing gate detecting interface featuring the chem‐SAM, the anti‐MUC1, and anti‐CD55 capturing antibodies and the b‐KRAS probe, along with the target markers. Also, the sketch of the gate coated with BSA serving as the negative‐control experiment is depicted on the right. In the other panels, the typical I D current shifts R I = (I – I 0 )/ I 0 are plotted against the cycling index for b) a high‐grade mucinous cyst, c) a low‐grade mucinous cyst, and (d) a non‐mucinous cyst. R I values are recorded at V GS and V D of −0.4 V, for each cycling index. The data points denote mean values derived from three replicates across three separate gates, with error bars calculated as relative standard deviations. The grey shaded area represents data collected post‐exposure to PBS reference fluid, while the red shading indicates measurements acquired following the exposure to the patient's specimen. The Limit‐of‐Identification (LOI) threshold is demarcated by a dashed‐dotted grey line.

Article Snippet: Monoclonal antibodies for Mucin 1 and the recombinant protein of human MUC1 were supplied by OriGene, while CD‐55 monoclonal antibodies and the human CD55 proteins were contributed by Abnova.

Techniques: Negative Control, Derivative Assay

Journal: Advanced Science

Article Title: Analysis of Clinical Samples of Pancreatic Cyst's Lesions with A Multi‐Analyte Bioelectronic Simot Array Benchmarked Against Ultrasensitive Chemiluminescent Immunoassay

doi: 10.1002/advs.202308141

Figure Lengend Snippet: a) Image of the SIMOA SP‐X 96‐well ELISA plate. Workflow illustrating the SIMOA SP‐X assay designed for the detection of b) MUC1 and c) CD55 target analytes. Calibration curves for d) MUC1 and e) CD55 assays recorded with 0.1 µg mL −1 of capture antibodies (either anti‐MUC1 or anti‐CD55) and 5 µg mL −1 of detector antibodies. MUC1 and CD55 standard solutions, prepared in PBS, were assayed, and presented as cyan and green hollow circles, respectively. In both cases, the blank is shown as black hollow circle. The sensing experiments were conducted in triplicates, while 6 blank replicates were acquired. The latter served to establish the Limit‐of‐Identification (LOI) achieved with the SIMOA assays. The modeling (solid curves) is performed using a logistic equation. The solid stars correspond to the SIMOA assay of cyst fluids from the high‐grade mucinous cyst SbU44, the low‐grade mucinous cyst SbU47, and the non‐mucinous cyst SbE8. Violin plots depict the distributions of concentrations for f) MUC1 and g) CD55 determined with the SIMOA assay for the 39 patients' samples.

Article Snippet: Monoclonal antibodies for Mucin 1 and the recombinant protein of human MUC1 were supplied by OriGene, while CD‐55 monoclonal antibodies and the human CD55 proteins were contributed by Abnova.

Techniques: Enzyme-linked Immunosorbent Assay