Journal: Cellular and Molecular Bioengineering

Article Title: Re-engineering Antimicrobial Peptides into Oncolytics Targeting Drug-Resistant Ovarian Cancers

doi: 10.1007/s12195-020-00626-z

Figure Lengend Snippet: Chemotherapeutic synergy. (a) Isobolograms of MAD1 and Doxorubicin (Dox), Paclitaxel (Ptx) or Cisplatin (Cis) combinatorial synergy in OVCAR-3 (left) and NCI/ADR-RES (right) cells. Fractional inhibitory concentration (FIC) < 1 and < 0.5 represent additive and synergistic effects, respectively. (b) Comparison of Dox and Cis IC50 towards NCI/ADR-RES as either a monotherapy (ADR-RES, black) or in combination with 20 μM MAD1 (ADR-RES + MAD1, grey). Activity of each drug as a monotherapy in pre-resistant OVCAR-3 (OVCAR-3, white) cells shown for comparison. (c–f) Indicated ovarian cancer cell line or patient-derived ovarian carcinoma cells were cultured in ultra-low attachment conditions and treated for 48 h with 2 μM cisplatin or 4 μM MAD1 alone and in combination. Data represent volume in arbitrary units (a.u.) and median.

Article Snippet: 3-(4,5-Dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT), Fmoc-protected amino acids, N,N 0 -diisopropylcarbodiimide (DIC), O-(7-Azabenzotriazol-1-yl)-N,N,N’,N’-tetramethyluronium hexafluorophosphate (HATU), N,N -dimethylformamide (DMF), and Paclitaxel were purchased from Chem-Impex International, Inc. RPMI-1640, M199, trypsin and EDTA solution, Antibiotic-Antimycotic Solution, ultra-low attachment (ULA) plates, and 1× phosphate buffered saline (PBS) were purchased from Corning.

Techniques: Concentration Assay, Comparison, Activity Assay, Derivative Assay, Cell Culture

Journal: Micromachines

Article Title: A Nitrocellulose Paper-Based Multi-Well Plate for Point-of-Care ELISA

doi: 10.3390/mi13122232

Figure Lengend Snippet: The schematic representation of the ( a ) direct ELISA of rabbit IgG and ( b ) the sandwich ELISA of sperm protein SP-10 on the NC paper multi-well plate.

Article Snippet: The sandwich ELISA protocol is described as follows: The sandwich ELISA of SP-10 starts with the addition of 3 μL of the capture antibody solution (mouse anti-ACRV1 monoclonal antibody in CBS solution, cat #11789-MM01, Sino Biological) with a concentration of 100 μg/mL, onto the NC paper well, followed by the overnight incubation in a 4 °C fridge for the sufficient antibody immobilization, Then, after taking the blocked plate out of the fridge, each NC paper well was pipetted with 10 μL of the wash buffer (0.05% Tween20 in TBS, pH 7.2–7.4) and blotted with an absorbent pad, three times, to remove the unbound antibodies, Next, 3 μL of the blocking buffer (2% BSA in wash buffer) was added to each well ( (b-i)), followed by a 10-min incubation and washing step with 10 μL wash buffer, three times, again to remove the unbound blocking reagents ( (b-ii)), Next, 3 μL of the SP-10 protein solution (cat #10227-H08B, Sino Biological) was added to the blocked NC paper well and incubated for 10 min, for the SP-10 protein to be captured by the immobilized capture antibody probe ( (b-iii)).

Techniques: Direct ELISA, Sandwich ELISA

Journal: Micromachines

Article Title: A Nitrocellulose Paper-Based Multi-Well Plate for Point-of-Care ELISA

doi: 10.3390/mi13122232

Figure Lengend Snippet: The comparison between the SP-10 NC-paper-based ELISA and the standard plate-based ELISA for the SP-10 detection.

Article Snippet: The sandwich ELISA protocol is described as follows: The sandwich ELISA of SP-10 starts with the addition of 3 μL of the capture antibody solution (mouse anti-ACRV1 monoclonal antibody in CBS solution, cat #11789-MM01, Sino Biological) with a concentration of 100 μg/mL, onto the NC paper well, followed by the overnight incubation in a 4 °C fridge for the sufficient antibody immobilization, Then, after taking the blocked plate out of the fridge, each NC paper well was pipetted with 10 μL of the wash buffer (0.05% Tween20 in TBS, pH 7.2–7.4) and blotted with an absorbent pad, three times, to remove the unbound antibodies, Next, 3 μL of the blocking buffer (2% BSA in wash buffer) was added to each well ( (b-i)), followed by a 10-min incubation and washing step with 10 μL wash buffer, three times, again to remove the unbound blocking reagents ( (b-ii)), Next, 3 μL of the SP-10 protein solution (cat #10227-H08B, Sino Biological) was added to the blocked NC paper well and incubated for 10 min, for the SP-10 protein to be captured by the immobilized capture antibody probe ( (b-iii)).

Techniques: Enzyme-linked Immunosorbent Assay, Sandwich ELISA, Biomarker Assay