In Vivo Imaging Sample Image - Day 7 - White Light - Raw

This file is a raw jpeg generated for Fig 9. It depicts imaging of a mouse in group 3 imaged under white light as described in the methods section

In Vitro Fluorescent Evaluation - CEA Internalization Data

This file contains the compiled and processed data surrounding the quantitative estimation of CEA internalization (Fig 5, panel B)

Immunofluorescence Sample Image BxPC-3 CEA 680 PTX

This file is a raw jpeg of an immunofluorescent image used in Fig 3, panel A

Conjugate Characterization - Data and Graphs

This file contains HPLC chromatograms and compiled numerical data concerning the characterization (drug:ab and fluorophore:ab ratios) of conjugates synthesized in this study (Fig 1 and Table 1)

In Vitro Fluorescent Evaluation - HeLa Flow Data

This file contains the raw flow cytometry data and overlayed images (“analyze” tab) which were arranged for Fig 4

Immunofluorescence Sample Image BxPC3 IgG 680

This file is a raw jpeg of an immunofluorescent image used in Fig 3, panel A

Conjugate Characterization - Linker Stability

This file contains all HPLC chromatograms, graphs and computations for determining antibody-drug linker stability of the fluorescent antibody drug conjugate as described in the paper (Fig 2)

MTT Cytotoxic Assessment of Samples and Conjugates.

<p>BxPC-3 cells (5x10³) were plated in 96-well collagen-coated plates, and treated with a range of various test samples as noted in the figure. Connected dashes denote conjugates, and “+” indicates the addition of free, unconjugated compound (i.e. α-CEA-680 is the fluorescent antibody conjugate, where α-CEA + 680 indicates naked antibody plus free fluorescent dye). The sample concentration at each data point is plotted according to the three x-axes. For example, the data point on the far right for α-CEA-680-PTX is comprised of 60ug/mL of antibody, which corresponds to an effective paclitaxel concentration of 400nM, and an effective DyLight 680 concentration of 800nM. Samples are diluted 1:2 serially throughout the rest of the series. Samples that do not contain antibody, paclitaxel, or DyLight 680 are still standardized in this manner for graphical comparison (i.e. the far right data point for naked antibody, α-CEA, is comprised of a 60ug/mL solution and serially diluted 1:2 as before, and α-CEA-PTX starts at 60ug/mL, which corresponds to an effective paclitaxel concentration of 400nM, and is serially diluted identically.) After 24 hours, samples were removed and replaced with fresh media. Cells were allowed to grow for 72 additional hours and then stained with MTT according to the manufacturer’s instructions. The absorbance of each well was measured at 540nm on the Thermo Scientific VarioSkan Flash plate spectrophotometer. Edges of the plate were omitted. Percent viability was defined as the ratio of the A540 of test wells compared to the A540 of untreated control wells. Untreated controls received media and an appropriate vehicle depending on sample. Data points represent the average of three separate experiments performed in triplicate (n = 9 wells). Error bars denote standard error.</p

IC₅₀ Estimations of Samples and Conjugates.

<p>IC₅₀ Estimations of Samples and Conjugates.</p

In Vivo Evaluation - Tumor Growth and Auto Exposure Data

This excel file contains all raw tumor growth measurements and graphs for in vivo efficacy evaluation (Fig 8). This file also contains the compiled exposure times generated for Fig 10