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nonlinear regression curve fitting package (sigmoidal dose–response)  (GraphPad Software Inc)


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    Structured Review

    GraphPad Software Inc nonlinear regression curve fitting package (sigmoidal dose–response)
    IC 50 and IC 90 values (μM) of anti-tumor drugs for monolayer and spheroid-cultured cells in a 96-well format (Fig. <xref ref-type= S2 , n = 3). Each cell culture was performed at 37 °C in an atmosphere containing 5% CO 2 , and treated with various concentrations of drugs for 48 h. Cell viability was assessed using the MTS assay reagent (Promega). Data were analyzed using a nonlinear regression curve-fitting package in Prism 4 (GraphPad Software Inc.) (Fig. S2 ). The drugs tested included: docetaxel, a mitotic inhibitor; irinotecan, a topoisomerase I inhibitor; 5-fluorouracil (5-FU), a thymidylate synthase inhibitor; sunitinib, a multi-receptor tyrosine kinase (RTK) inhibitor; tirapazamine, a hypoxia-activated prodrug; and FX11, a lactate dehydrogenase inhibitor. Radial organization of each cell on the spheroidal culture dish (IWAKI) conferred resistance to the most of drugs tested." width="250" height="auto" />
    Nonlinear Regression Curve Fitting Package (Sigmoidal Dose–Response), supplied by GraphPad Software Inc, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/nonlinear+regression+(curve+fit)-boltzmann+sigmoidal/nonlinear+regression+curve+fitting+package++sigmoidal+dose+response+/pmc12130504-157-19-29
    Average 90 stars, based on 1 article reviews
    nonlinear regression curve fitting package (sigmoidal dose–response) - by Bioz Stars, 2026-09
    90/100 stars

    Images

    1) Product Images from "Hyperpolarized [1- 13 C]pyruvate NMR spectroscopy reveals transition of tumor energy metabolism in microscale multicellular spheroids"

    Article Title: Hyperpolarized [1- 13 C]pyruvate NMR spectroscopy reveals transition of tumor energy metabolism in microscale multicellular spheroids

    Journal: Scientific Reports

    doi: 10.1038/s41598-025-03454-1

    S2 , n = 3). Each cell culture was performed at 37 °C in an atmosphere containing 5% CO 2 , and treated with various concentrations of drugs for 48 h. Cell viability was assessed using the MTS assay reagent (Promega). Data were analyzed using a nonlinear regression curve-fitting package in Prism 4 (GraphPad Software Inc.) (Fig. S2 ). The drugs tested included: docetaxel, a mitotic inhibitor; irinotecan, a topoisomerase I inhibitor; 5-fluorouracil (5-FU), a thymidylate synthase inhibitor; sunitinib, a multi-receptor tyrosine kinase (RTK) inhibitor; tirapazamine, a hypoxia-activated prodrug; and FX11, a lactate dehydrogenase inhibitor. Radial organization of each cell on the spheroidal culture dish (IWAKI) conferred resistance to the most of drugs tested." title="... assay reagent (Promega). Data were analyzed using a nonlinear regression curve-fitting package in Prism 4 ..." property="contentUrl" width="100%" height="100%"/>
    Figure Legend Snippet: IC 50 and IC 90 values (μM) of anti-tumor drugs for monolayer and spheroid-cultured cells in a 96-well format (Fig. S2 , n = 3). Each cell culture was performed at 37 °C in an atmosphere containing 5% CO 2 , and treated with various concentrations of drugs for 48 h. Cell viability was assessed using the MTS assay reagent (Promega). Data were analyzed using a nonlinear regression curve-fitting package in Prism 4 (GraphPad Software Inc.) (Fig. S2 ). The drugs tested included: docetaxel, a mitotic inhibitor; irinotecan, a topoisomerase I inhibitor; 5-fluorouracil (5-FU), a thymidylate synthase inhibitor; sunitinib, a multi-receptor tyrosine kinase (RTK) inhibitor; tirapazamine, a hypoxia-activated prodrug; and FX11, a lactate dehydrogenase inhibitor. Radial organization of each cell on the spheroidal culture dish (IWAKI) conferred resistance to the most of drugs tested.

    Techniques Used: Cell Culture, MTS Assay, Software

    Related Articles

    MTT Assay:

    Article Title: Antibody-drug conjugates comprising anti-B7-H3 antibodies
    Article Snippet: IC50 was generated using a sigmoidal dose-response nonlinear regression curve fit (GraphPad software Inc.) and the results are shown in FIGS. 1-9 and Tables 17-27 below.

    Article Title: Structural basis for saxitoxin congener binding and neutralization by anuran saxiphilins.
    Article Snippet: The half maximal effective concentrations (EC50) were calculated by fitting normalized counts per minute as a function of toxin concentration using the nonlinear regression sigmoidal dose-response (variable slope) curve fit with weight by 1/Y2 and bottoms constrained to a shared value for all data sets in GraphPad Prism.

    Article Title: Benzodiazepine derivatives and uses thereof
    Article Snippet: The reaction mixture was purified by preparative HPLC (Column: Innoval ODS-2 10 um, 100 Å, 21.2×250 mm; flow rate: 15 mL/min, A buffer 0.1% Formic acid in water/B buffer 0.1% Formic acid in ACN, method gradient, solvent A:solvent B 95:5 to 5:95, 1 hour, wavelength 214 nm) to obtain compound T-Int-7 (44.4 mg, 82%) as ivory solid.

    Article Title: Substituted benzo[5,6][1,4]diazepino[1,2-a]indoles for the treatment of proliferative disorders
    Article Snippet: Killing curves and IC50 were generated using a sigmoidal dose-response nonlinear regression curve fit (GraphPad Software Inc.).

    Article Title: Substituted benzo[5,6][1,4]diazepino[1,2-a]indoles for the treatment of proliferative disorders
    Article Snippet: Killing curves and IC50 were generated using a sigmoidal dose-response nonlinear regression curve fit (GraphPad Software Inc.) TABLE 8 In vitro bystander IC50 (Molar, M) in human tumor cells, −/ + FRα FRα-negative Namalwa-Luc alone FRα-positive + Antibody- Compound (non-specific Namalwa-Luc mixed linker # activity of ADC) KB OV-90 JEG-3 T47D M-sSPDB 17 4e−9 6e−11 9e−11 7e−10 7e−11 M-sSPDB 109 2e−9 3e−9 2e−9 2e−9 ND M- 160 1e−8 3e−10 1e−8 3e−9 ND M- 166 8e−9 3e−10 ND 2e−9 ND M-sSPDB 208 2e−10 3e−11 2e−10 ND 3e−11 M-sSPDB 204 1e−9 4e−12 4e−10 ND 4e−11 ND = not determined

    Article Title: Structural basis for saxitoxin congener binding and neutralization by anuran saxiphilins
    Article Snippet: The half maximal effective concentrations (EC 50 ) were calculated by fitting normalized counts per minute as a function of toxin concentration using the nonlinear regression sigmoidal dose-response (variable slope) curve fit with weight by 1/Y 2 and bottoms constrained to a shared value for all data sets in GraphPad Prism.

    Article Title: Substituted benzo[5,6][1,4]diazepino[1,2-a]indoles for the treatment of proliferative disorders
    Article Snippet: Binding curves and EC50 were generated using a sigmoidal dose-response nonlinear regression curve fit (GraphPad Software Inc.) TABLE 9 EC50 values (Molar, M) determined by in vitro binding assays using flow cytometry Antibody- Un-conjugated linker Compound # Conjugate Antibody control* M- 10 3.00E−10 3.00E−10 M-sSPDB 17 2.00E−10 2.00E−10 M-sSPDB 30 3.00E−10 2.00E−10 M-sSPDB 87 4.00E−10 2.00E−10 M-sSPDB 109 3.00E−10 2.00E−10 M-sSPDB 127 2.00E−10 2.00E−10 M- 143 2.00E−10 1.00E−10 M-sSPDB 155 3.00E−10 1.00E−10 M- 160 2.00E−10 2.00E−10 M- 166 2.00E−10 2.00E−10 M-sSPDB 204 6.00E−10 5.00E−10 M-sSPDB 208 7.00E−10 5.00E−10 *The EC50 values for each conjugate and the unconjugated antibody control were generated in independent experiments which might explain slight variability of the unconjugated control antibody EC50 values.

    Article Title: Substituted benzodiazepines as antibody-drug conjugates
    Article Snippet: Cell viability was determined by the MTT assay and IC50 was generated using a sigmoidal dose-response nonlinear regression curve fit (GraphPad software Inc.) Results of in vitro analysis of compounds T-1-AB, T-2-AB, T-3-AB, and T-4-AB, are shown in FIG. 4, FIG. 5, FIG. 6, FIG. 7, and Table 4.

    Generated:

    Article Title: Antibody-drug conjugates comprising anti-B7-H3 antibodies
    Article Snippet: IC50 was generated using a sigmoidal dose-response nonlinear regression curve fit (GraphPad software Inc.) and the results are shown in FIGS. 1-9 and Tables 17-27 below.

    Article Title: Structural basis for saxitoxin congener binding and neutralization by anuran saxiphilins.
    Article Snippet: The half maximal effective concentrations (EC50) were calculated by fitting normalized counts per minute as a function of toxin concentration using the nonlinear regression sigmoidal dose-response (variable slope) curve fit with weight by 1/Y2 and bottoms constrained to a shared value for all data sets in GraphPad Prism.

    Article Title: Benzodiazepine derivatives and uses thereof
    Article Snippet: The reaction mixture was purified by preparative HPLC (Column: Innoval ODS-2 10 um, 100 Å, 21.2×250 mm; flow rate: 15 mL/min, A buffer 0.1% Formic acid in water/B buffer 0.1% Formic acid in ACN, method gradient, solvent A:solvent B 95:5 to 5:95, 1 hour, wavelength 214 nm) to obtain compound T-Int-7 (44.4 mg, 82%) as ivory solid.

    Article Title: Substituted benzo[5,6][1,4]diazepino[1,2-a]indoles for the treatment of proliferative disorders
    Article Snippet: Killing curves and IC50 were generated using a sigmoidal dose-response nonlinear regression curve fit (GraphPad Software Inc.).

    Article Title: Substituted benzo[5,6][1,4]diazepino[1,2-a]indoles for the treatment of proliferative disorders
    Article Snippet: Killing curves and IC50 were generated using a sigmoidal dose-response nonlinear regression curve fit (GraphPad Software Inc.) TABLE 8 In vitro bystander IC50 (Molar, M) in human tumor cells, −/ + FRα FRα-negative Namalwa-Luc alone FRα-positive + Antibody- Compound (non-specific Namalwa-Luc mixed linker # activity of ADC) KB OV-90 JEG-3 T47D M-sSPDB 17 4e−9 6e−11 9e−11 7e−10 7e−11 M-sSPDB 109 2e−9 3e−9 2e−9 2e−9 ND M- 160 1e−8 3e−10 1e−8 3e−9 ND M- 166 8e−9 3e−10 ND 2e−9 ND M-sSPDB 208 2e−10 3e−11 2e−10 ND 3e−11 M-sSPDB 204 1e−9 4e−12 4e−10 ND 4e−11 ND = not determined

    Article Title: Structural basis for saxitoxin congener binding and neutralization by anuran saxiphilins
    Article Snippet: The half maximal effective concentrations (EC 50 ) were calculated by fitting normalized counts per minute as a function of toxin concentration using the nonlinear regression sigmoidal dose-response (variable slope) curve fit with weight by 1/Y 2 and bottoms constrained to a shared value for all data sets in GraphPad Prism.

    Article Title: Substituted benzo[5,6][1,4]diazepino[1,2-a]indoles for the treatment of proliferative disorders
    Article Snippet: Binding curves and EC50 were generated using a sigmoidal dose-response nonlinear regression curve fit (GraphPad Software Inc.) TABLE 9 EC50 values (Molar, M) determined by in vitro binding assays using flow cytometry Antibody- Un-conjugated linker Compound # Conjugate Antibody control* M- 10 3.00E−10 3.00E−10 M-sSPDB 17 2.00E−10 2.00E−10 M-sSPDB 30 3.00E−10 2.00E−10 M-sSPDB 87 4.00E−10 2.00E−10 M-sSPDB 109 3.00E−10 2.00E−10 M-sSPDB 127 2.00E−10 2.00E−10 M- 143 2.00E−10 1.00E−10 M-sSPDB 155 3.00E−10 1.00E−10 M- 160 2.00E−10 2.00E−10 M- 166 2.00E−10 2.00E−10 M-sSPDB 204 6.00E−10 5.00E−10 M-sSPDB 208 7.00E−10 5.00E−10 *The EC50 values for each conjugate and the unconjugated antibody control were generated in independent experiments which might explain slight variability of the unconjugated control antibody EC50 values.

    Article Title: Substituted benzodiazepines as antibody-drug conjugates
    Article Snippet: Cell viability was determined by the MTT assay and IC50 was generated using a sigmoidal dose-response nonlinear regression curve fit (GraphPad software Inc.) Results of in vitro analysis of compounds T-1-AB, T-2-AB, T-3-AB, and T-4-AB, are shown in FIG. 4, FIG. 5, FIG. 6, FIG. 7, and Table 4.

    In Vitro:

    Article Title: Antibody-drug conjugates comprising anti-B7-H3 antibodies
    Article Snippet: IC50 was generated using a sigmoidal dose-response nonlinear regression curve fit (GraphPad software Inc.) and the results are shown in FIGS. 1-9 and Tables 17-27 below.

    Article Title: Structural basis for saxitoxin congener binding and neutralization by anuran saxiphilins.
    Article Snippet: The half maximal effective concentrations (EC50) were calculated by fitting normalized counts per minute as a function of toxin concentration using the nonlinear regression sigmoidal dose-response (variable slope) curve fit with weight by 1/Y2 and bottoms constrained to a shared value for all data sets in GraphPad Prism.

    Article Title: Benzodiazepine derivatives and uses thereof
    Article Snippet: The reaction mixture was purified by preparative HPLC (Column: Innoval ODS-2 10 um, 100 Å, 21.2×250 mm; flow rate: 15 mL/min, A buffer 0.1% Formic acid in water/B buffer 0.1% Formic acid in ACN, method gradient, solvent A:solvent B 95:5 to 5:95, 1 hour, wavelength 214 nm) to obtain compound T-Int-7 (44.4 mg, 82%) as ivory solid.

    Article Title: Substituted benzo[5,6][1,4]diazepino[1,2-a]indoles for the treatment of proliferative disorders
    Article Snippet: Killing curves and IC50 were generated using a sigmoidal dose-response nonlinear regression curve fit (GraphPad Software Inc.).

    Article Title: Substituted benzo[5,6][1,4]diazepino[1,2-a]indoles for the treatment of proliferative disorders
    Article Snippet: Killing curves and IC50 were generated using a sigmoidal dose-response nonlinear regression curve fit (GraphPad Software Inc.) TABLE 8 In vitro bystander IC50 (Molar, M) in human tumor cells, −/ + FRα FRα-negative Namalwa-Luc alone FRα-positive + Antibody- Compound (non-specific Namalwa-Luc mixed linker # activity of ADC) KB OV-90 JEG-3 T47D M-sSPDB 17 4e−9 6e−11 9e−11 7e−10 7e−11 M-sSPDB 109 2e−9 3e−9 2e−9 2e−9 ND M- 160 1e−8 3e−10 1e−8 3e−9 ND M- 166 8e−9 3e−10 ND 2e−9 ND M-sSPDB 208 2e−10 3e−11 2e−10 ND 3e−11 M-sSPDB 204 1e−9 4e−12 4e−10 ND 4e−11 ND = not determined

    Article Title: Structural basis for saxitoxin congener binding and neutralization by anuran saxiphilins
    Article Snippet: The half maximal effective concentrations (EC 50 ) were calculated by fitting normalized counts per minute as a function of toxin concentration using the nonlinear regression sigmoidal dose-response (variable slope) curve fit with weight by 1/Y 2 and bottoms constrained to a shared value for all data sets in GraphPad Prism.

    Article Title: Substituted benzo[5,6][1,4]diazepino[1,2-a]indoles for the treatment of proliferative disorders
    Article Snippet: Binding curves and EC50 were generated using a sigmoidal dose-response nonlinear regression curve fit (GraphPad Software Inc.) TABLE 9 EC50 values (Molar, M) determined by in vitro binding assays using flow cytometry Antibody- Un-conjugated linker Compound # Conjugate Antibody control* M- 10 3.00E−10 3.00E−10 M-sSPDB 17 2.00E−10 2.00E−10 M-sSPDB 30 3.00E−10 2.00E−10 M-sSPDB 87 4.00E−10 2.00E−10 M-sSPDB 109 3.00E−10 2.00E−10 M-sSPDB 127 2.00E−10 2.00E−10 M- 143 2.00E−10 1.00E−10 M-sSPDB 155 3.00E−10 1.00E−10 M- 160 2.00E−10 2.00E−10 M- 166 2.00E−10 2.00E−10 M-sSPDB 204 6.00E−10 5.00E−10 M-sSPDB 208 7.00E−10 5.00E−10 *The EC50 values for each conjugate and the unconjugated antibody control were generated in independent experiments which might explain slight variability of the unconjugated control antibody EC50 values.

    Article Title: Substituted benzodiazepines as antibody-drug conjugates
    Article Snippet: Cell viability was determined by the MTT assay and IC50 was generated using a sigmoidal dose-response nonlinear regression curve fit (GraphPad software Inc.) Results of in vitro analysis of compounds T-1-AB, T-2-AB, T-3-AB, and T-4-AB, are shown in FIG. 4, FIG. 5, FIG. 6, FIG. 7, and Table 4.

    Activity Assay:

    Article Title: Antibody-drug conjugates comprising anti-B7-H3 antibodies
    Article Snippet: IC50 was generated using a sigmoidal dose-response nonlinear regression curve fit (GraphPad software Inc.) and the results are shown in FIGS. 1-9 and Tables 17-27 below.

    Article Title: Structural basis for saxitoxin congener binding and neutralization by anuran saxiphilins.
    Article Snippet: The half maximal effective concentrations (EC50) were calculated by fitting normalized counts per minute as a function of toxin concentration using the nonlinear regression sigmoidal dose-response (variable slope) curve fit with weight by 1/Y2 and bottoms constrained to a shared value for all data sets in GraphPad Prism.

    Article Title: Benzodiazepine derivatives and uses thereof
    Article Snippet: The reaction mixture was purified by preparative HPLC (Column: Innoval ODS-2 10 um, 100 Å, 21.2×250 mm; flow rate: 15 mL/min, A buffer 0.1% Formic acid in water/B buffer 0.1% Formic acid in ACN, method gradient, solvent A:solvent B 95:5 to 5:95, 1 hour, wavelength 214 nm) to obtain compound T-Int-7 (44.4 mg, 82%) as ivory solid.

    Article Title: Substituted benzo[5,6][1,4]diazepino[1,2-a]indoles for the treatment of proliferative disorders
    Article Snippet: Killing curves and IC50 were generated using a sigmoidal dose-response nonlinear regression curve fit (GraphPad Software Inc.).

    Article Title: Substituted benzo[5,6][1,4]diazepino[1,2-a]indoles for the treatment of proliferative disorders
    Article Snippet: Killing curves and IC50 were generated using a sigmoidal dose-response nonlinear regression curve fit (GraphPad Software Inc.) TABLE 8 In vitro bystander IC50 (Molar, M) in human tumor cells, −/ + FRα FRα-negative Namalwa-Luc alone FRα-positive + Antibody- Compound (non-specific Namalwa-Luc mixed linker # activity of ADC) KB OV-90 JEG-3 T47D M-sSPDB 17 4e−9 6e−11 9e−11 7e−10 7e−11 M-sSPDB 109 2e−9 3e−9 2e−9 2e−9 ND M- 160 1e−8 3e−10 1e−8 3e−9 ND M- 166 8e−9 3e−10 ND 2e−9 ND M-sSPDB 208 2e−10 3e−11 2e−10 ND 3e−11 M-sSPDB 204 1e−9 4e−12 4e−10 ND 4e−11 ND = not determined

    Article Title: Structural basis for saxitoxin congener binding and neutralization by anuran saxiphilins
    Article Snippet: The half maximal effective concentrations (EC 50 ) were calculated by fitting normalized counts per minute as a function of toxin concentration using the nonlinear regression sigmoidal dose-response (variable slope) curve fit with weight by 1/Y 2 and bottoms constrained to a shared value for all data sets in GraphPad Prism.

    Article Title: Substituted benzo[5,6][1,4]diazepino[1,2-a]indoles for the treatment of proliferative disorders
    Article Snippet: Binding curves and EC50 were generated using a sigmoidal dose-response nonlinear regression curve fit (GraphPad Software Inc.) TABLE 9 EC50 values (Molar, M) determined by in vitro binding assays using flow cytometry Antibody- Un-conjugated linker Compound # Conjugate Antibody control* M- 10 3.00E−10 3.00E−10 M-sSPDB 17 2.00E−10 2.00E−10 M-sSPDB 30 3.00E−10 2.00E−10 M-sSPDB 87 4.00E−10 2.00E−10 M-sSPDB 109 3.00E−10 2.00E−10 M-sSPDB 127 2.00E−10 2.00E−10 M- 143 2.00E−10 1.00E−10 M-sSPDB 155 3.00E−10 1.00E−10 M- 160 2.00E−10 2.00E−10 M- 166 2.00E−10 2.00E−10 M-sSPDB 204 6.00E−10 5.00E−10 M-sSPDB 208 7.00E−10 5.00E−10 *The EC50 values for each conjugate and the unconjugated antibody control were generated in independent experiments which might explain slight variability of the unconjugated control antibody EC50 values.

    Article Title: Substituted benzodiazepines as antibody-drug conjugates
    Article Snippet: Cell viability was determined by the MTT assay and IC50 was generated using a sigmoidal dose-response nonlinear regression curve fit (GraphPad software Inc.) Results of in vitro analysis of compounds T-1-AB, T-2-AB, T-3-AB, and T-4-AB, are shown in FIG. 4, FIG. 5, FIG. 6, FIG. 7, and Table 4.

    Binding Assay:

    Article Title: Antibody-drug conjugates comprising anti-B7-H3 antibodies
    Article Snippet: IC50 was generated using a sigmoidal dose-response nonlinear regression curve fit (GraphPad software Inc.) and the results are shown in FIGS. 1-9 and Tables 17-27 below.

    Article Title: Structural basis for saxitoxin congener binding and neutralization by anuran saxiphilins.
    Article Snippet: The half maximal effective concentrations (EC50) were calculated by fitting normalized counts per minute as a function of toxin concentration using the nonlinear regression sigmoidal dose-response (variable slope) curve fit with weight by 1/Y2 and bottoms constrained to a shared value for all data sets in GraphPad Prism.

    Article Title: Benzodiazepine derivatives and uses thereof
    Article Snippet: The reaction mixture was purified by preparative HPLC (Column: Innoval ODS-2 10 um, 100 Å, 21.2×250 mm; flow rate: 15 mL/min, A buffer 0.1% Formic acid in water/B buffer 0.1% Formic acid in ACN, method gradient, solvent A:solvent B 95:5 to 5:95, 1 hour, wavelength 214 nm) to obtain compound T-Int-7 (44.4 mg, 82%) as ivory solid.

    Article Title: Substituted benzo[5,6][1,4]diazepino[1,2-a]indoles for the treatment of proliferative disorders
    Article Snippet: Killing curves and IC50 were generated using a sigmoidal dose-response nonlinear regression curve fit (GraphPad Software Inc.).

    Article Title: Substituted benzo[5,6][1,4]diazepino[1,2-a]indoles for the treatment of proliferative disorders
    Article Snippet: Killing curves and IC50 were generated using a sigmoidal dose-response nonlinear regression curve fit (GraphPad Software Inc.) TABLE 8 In vitro bystander IC50 (Molar, M) in human tumor cells, −/ + FRα FRα-negative Namalwa-Luc alone FRα-positive + Antibody- Compound (non-specific Namalwa-Luc mixed linker # activity of ADC) KB OV-90 JEG-3 T47D M-sSPDB 17 4e−9 6e−11 9e−11 7e−10 7e−11 M-sSPDB 109 2e−9 3e−9 2e−9 2e−9 ND M- 160 1e−8 3e−10 1e−8 3e−9 ND M- 166 8e−9 3e−10 ND 2e−9 ND M-sSPDB 208 2e−10 3e−11 2e−10 ND 3e−11 M-sSPDB 204 1e−9 4e−12 4e−10 ND 4e−11 ND = not determined

    Article Title: Structural basis for saxitoxin congener binding and neutralization by anuran saxiphilins
    Article Snippet: The half maximal effective concentrations (EC 50 ) were calculated by fitting normalized counts per minute as a function of toxin concentration using the nonlinear regression sigmoidal dose-response (variable slope) curve fit with weight by 1/Y 2 and bottoms constrained to a shared value for all data sets in GraphPad Prism.

    Article Title: Substituted benzo[5,6][1,4]diazepino[1,2-a]indoles for the treatment of proliferative disorders
    Article Snippet: Binding curves and EC50 were generated using a sigmoidal dose-response nonlinear regression curve fit (GraphPad Software Inc.) TABLE 9 EC50 values (Molar, M) determined by in vitro binding assays using flow cytometry Antibody- Un-conjugated linker Compound # Conjugate Antibody control* M- 10 3.00E−10 3.00E−10 M-sSPDB 17 2.00E−10 2.00E−10 M-sSPDB 30 3.00E−10 2.00E−10 M-sSPDB 87 4.00E−10 2.00E−10 M-sSPDB 109 3.00E−10 2.00E−10 M-sSPDB 127 2.00E−10 2.00E−10 M- 143 2.00E−10 1.00E−10 M-sSPDB 155 3.00E−10 1.00E−10 M- 160 2.00E−10 2.00E−10 M- 166 2.00E−10 2.00E−10 M-sSPDB 204 6.00E−10 5.00E−10 M-sSPDB 208 7.00E−10 5.00E−10 *The EC50 values for each conjugate and the unconjugated antibody control were generated in independent experiments which might explain slight variability of the unconjugated control antibody EC50 values.

    Article Title: Substituted benzodiazepines as antibody-drug conjugates
    Article Snippet: Cell viability was determined by the MTT assay and IC50 was generated using a sigmoidal dose-response nonlinear regression curve fit (GraphPad software Inc.) Results of in vitro analysis of compounds T-1-AB, T-2-AB, T-3-AB, and T-4-AB, are shown in FIG. 4, FIG. 5, FIG. 6, FIG. 7, and Table 4.

    Flow Cytometry:

    Article Title: Antibody-drug conjugates comprising anti-B7-H3 antibodies
    Article Snippet: IC50 was generated using a sigmoidal dose-response nonlinear regression curve fit (GraphPad software Inc.) and the results are shown in FIGS. 1-9 and Tables 17-27 below.

    Article Title: Structural basis for saxitoxin congener binding and neutralization by anuran saxiphilins.
    Article Snippet: The half maximal effective concentrations (EC50) were calculated by fitting normalized counts per minute as a function of toxin concentration using the nonlinear regression sigmoidal dose-response (variable slope) curve fit with weight by 1/Y2 and bottoms constrained to a shared value for all data sets in GraphPad Prism.

    Article Title: Benzodiazepine derivatives and uses thereof
    Article Snippet: The reaction mixture was purified by preparative HPLC (Column: Innoval ODS-2 10 um, 100 Å, 21.2×250 mm; flow rate: 15 mL/min, A buffer 0.1% Formic acid in water/B buffer 0.1% Formic acid in ACN, method gradient, solvent A:solvent B 95:5 to 5:95, 1 hour, wavelength 214 nm) to obtain compound T-Int-7 (44.4 mg, 82%) as ivory solid.

    Article Title: Substituted benzo[5,6][1,4]diazepino[1,2-a]indoles for the treatment of proliferative disorders
    Article Snippet: Killing curves and IC50 were generated using a sigmoidal dose-response nonlinear regression curve fit (GraphPad Software Inc.).

    Article Title: Substituted benzo[5,6][1,4]diazepino[1,2-a]indoles for the treatment of proliferative disorders
    Article Snippet: Killing curves and IC50 were generated using a sigmoidal dose-response nonlinear regression curve fit (GraphPad Software Inc.) TABLE 8 In vitro bystander IC50 (Molar, M) in human tumor cells, −/ + FRα FRα-negative Namalwa-Luc alone FRα-positive + Antibody- Compound (non-specific Namalwa-Luc mixed linker # activity of ADC) KB OV-90 JEG-3 T47D M-sSPDB 17 4e−9 6e−11 9e−11 7e−10 7e−11 M-sSPDB 109 2e−9 3e−9 2e−9 2e−9 ND M- 160 1e−8 3e−10 1e−8 3e−9 ND M- 166 8e−9 3e−10 ND 2e−9 ND M-sSPDB 208 2e−10 3e−11 2e−10 ND 3e−11 M-sSPDB 204 1e−9 4e−12 4e−10 ND 4e−11 ND = not determined

    Article Title: Structural basis for saxitoxin congener binding and neutralization by anuran saxiphilins
    Article Snippet: The half maximal effective concentrations (EC 50 ) were calculated by fitting normalized counts per minute as a function of toxin concentration using the nonlinear regression sigmoidal dose-response (variable slope) curve fit with weight by 1/Y 2 and bottoms constrained to a shared value for all data sets in GraphPad Prism.

    Article Title: Substituted benzo[5,6][1,4]diazepino[1,2-a]indoles for the treatment of proliferative disorders
    Article Snippet: Binding curves and EC50 were generated using a sigmoidal dose-response nonlinear regression curve fit (GraphPad Software Inc.) TABLE 9 EC50 values (Molar, M) determined by in vitro binding assays using flow cytometry Antibody- Un-conjugated linker Compound # Conjugate Antibody control* M- 10 3.00E−10 3.00E−10 M-sSPDB 17 2.00E−10 2.00E−10 M-sSPDB 30 3.00E−10 2.00E−10 M-sSPDB 87 4.00E−10 2.00E−10 M-sSPDB 109 3.00E−10 2.00E−10 M-sSPDB 127 2.00E−10 2.00E−10 M- 143 2.00E−10 1.00E−10 M-sSPDB 155 3.00E−10 1.00E−10 M- 160 2.00E−10 2.00E−10 M- 166 2.00E−10 2.00E−10 M-sSPDB 204 6.00E−10 5.00E−10 M-sSPDB 208 7.00E−10 5.00E−10 *The EC50 values for each conjugate and the unconjugated antibody control were generated in independent experiments which might explain slight variability of the unconjugated control antibody EC50 values.

    Article Title: Substituted benzodiazepines as antibody-drug conjugates
    Article Snippet: Cell viability was determined by the MTT assay and IC50 was generated using a sigmoidal dose-response nonlinear regression curve fit (GraphPad software Inc.) Results of in vitro analysis of compounds T-1-AB, T-2-AB, T-3-AB, and T-4-AB, are shown in FIG. 4, FIG. 5, FIG. 6, FIG. 7, and Table 4.

    Control:

    Article Title: Antibody-drug conjugates comprising anti-B7-H3 antibodies
    Article Snippet: IC50 was generated using a sigmoidal dose-response nonlinear regression curve fit (GraphPad software Inc.) and the results are shown in FIGS. 1-9 and Tables 17-27 below.

    Article Title: Structural basis for saxitoxin congener binding and neutralization by anuran saxiphilins.
    Article Snippet: The half maximal effective concentrations (EC50) were calculated by fitting normalized counts per minute as a function of toxin concentration using the nonlinear regression sigmoidal dose-response (variable slope) curve fit with weight by 1/Y2 and bottoms constrained to a shared value for all data sets in GraphPad Prism.

    Article Title: Benzodiazepine derivatives and uses thereof
    Article Snippet: The reaction mixture was purified by preparative HPLC (Column: Innoval ODS-2 10 um, 100 Å, 21.2×250 mm; flow rate: 15 mL/min, A buffer 0.1% Formic acid in water/B buffer 0.1% Formic acid in ACN, method gradient, solvent A:solvent B 95:5 to 5:95, 1 hour, wavelength 214 nm) to obtain compound T-Int-7 (44.4 mg, 82%) as ivory solid.

    Article Title: Substituted benzo[5,6][1,4]diazepino[1,2-a]indoles for the treatment of proliferative disorders
    Article Snippet: Killing curves and IC50 were generated using a sigmoidal dose-response nonlinear regression curve fit (GraphPad Software Inc.).

    Article Title: Substituted benzo[5,6][1,4]diazepino[1,2-a]indoles for the treatment of proliferative disorders
    Article Snippet: Killing curves and IC50 were generated using a sigmoidal dose-response nonlinear regression curve fit (GraphPad Software Inc.) TABLE 8 In vitro bystander IC50 (Molar, M) in human tumor cells, −/ + FRα FRα-negative Namalwa-Luc alone FRα-positive + Antibody- Compound (non-specific Namalwa-Luc mixed linker # activity of ADC) KB OV-90 JEG-3 T47D M-sSPDB 17 4e−9 6e−11 9e−11 7e−10 7e−11 M-sSPDB 109 2e−9 3e−9 2e−9 2e−9 ND M- 160 1e−8 3e−10 1e−8 3e−9 ND M- 166 8e−9 3e−10 ND 2e−9 ND M-sSPDB 208 2e−10 3e−11 2e−10 ND 3e−11 M-sSPDB 204 1e−9 4e−12 4e−10 ND 4e−11 ND = not determined

    Article Title: Structural basis for saxitoxin congener binding and neutralization by anuran saxiphilins
    Article Snippet: The half maximal effective concentrations (EC 50 ) were calculated by fitting normalized counts per minute as a function of toxin concentration using the nonlinear regression sigmoidal dose-response (variable slope) curve fit with weight by 1/Y 2 and bottoms constrained to a shared value for all data sets in GraphPad Prism.

    Article Title: Substituted benzo[5,6][1,4]diazepino[1,2-a]indoles for the treatment of proliferative disorders
    Article Snippet: Binding curves and EC50 were generated using a sigmoidal dose-response nonlinear regression curve fit (GraphPad Software Inc.) TABLE 9 EC50 values (Molar, M) determined by in vitro binding assays using flow cytometry Antibody- Un-conjugated linker Compound # Conjugate Antibody control* M- 10 3.00E−10 3.00E−10 M-sSPDB 17 2.00E−10 2.00E−10 M-sSPDB 30 3.00E−10 2.00E−10 M-sSPDB 87 4.00E−10 2.00E−10 M-sSPDB 109 3.00E−10 2.00E−10 M-sSPDB 127 2.00E−10 2.00E−10 M- 143 2.00E−10 1.00E−10 M-sSPDB 155 3.00E−10 1.00E−10 M- 160 2.00E−10 2.00E−10 M- 166 2.00E−10 2.00E−10 M-sSPDB 204 6.00E−10 5.00E−10 M-sSPDB 208 7.00E−10 5.00E−10 *The EC50 values for each conjugate and the unconjugated antibody control were generated in independent experiments which might explain slight variability of the unconjugated control antibody EC50 values.

    Article Title: Substituted benzodiazepines as antibody-drug conjugates
    Article Snippet: Cell viability was determined by the MTT assay and IC50 was generated using a sigmoidal dose-response nonlinear regression curve fit (GraphPad software Inc.) Results of in vitro analysis of compounds T-1-AB, T-2-AB, T-3-AB, and T-4-AB, are shown in FIG. 4, FIG. 5, FIG. 6, FIG. 7, and Table 4.



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    Figure Lengend Snippet: IC 50 and IC 90 values (μM) of anti-tumor drugs for monolayer and spheroid-cultured cells in a 96-well format (Fig. S2 , n = 3). Each cell culture was performed at 37 °C in an atmosphere containing 5% CO 2 , and treated with various concentrations of drugs for 48 h. Cell viability was assessed using the MTS assay reagent (Promega). Data were analyzed using a nonlinear regression curve-fitting package in Prism 4 (GraphPad Software Inc.) (Fig. S2 ). The drugs tested included: docetaxel, a mitotic inhibitor; irinotecan, a topoisomerase I inhibitor; 5-fluorouracil (5-FU), a thymidylate synthase inhibitor; sunitinib, a multi-receptor tyrosine kinase (RTK) inhibitor; tirapazamine, a hypoxia-activated prodrug; and FX11, a lactate dehydrogenase inhibitor. Radial organization of each cell on the spheroidal culture dish (IWAKI) conferred resistance to the most of drugs tested.

    Article Snippet: Cell viability in each drug concentration was plotted (Fig. ), and the sigmoidal dose–response curve was calculated using a nonlinear regression curve fitting package (sigmoidal dose–response) in Prism 4 (GraphPad Software Inc., La Jolla, CA).

    Techniques: Cell Culture, MTS Assay, Software