Journal: bioRxiv

Article Title: Anti-HIV Immunotoxin and Antibody-Drug Conjugate Display Both Common and Distinct Effects in Killing Target Cells

doi: 10.64898/2026.04.07.717054

Figure Lengend Snippet: A . The binding of anti-gp41 7B2 to H9/N:L4-3 cells was demonstrated by indirect immunofluorescence and flow cytometry. The gray histogram indicates binding in the presence of secondary FITC-conjugated goat anti-human IgG only, the blue histogram demonstrates fluorescence of cells first incubated with 7B2 followed by FITC-conjugated secondary antibody. B. Comparative cytotoxicity of 7B2-dgA and 7B2-PNU. Cells were incubated with the indicated concentration of CIC and CD4-IgG2 (500 ng/mL) for 72 hr. MTS dye reduction was measured during the final 3 hr of culture. C. The kinetics of apoptosis induction by the CICs was measured by the binding of fluorescent annexin V to cells at different times following treatment with CIC.

Article Snippet: Cells were washed twice in PBS and incubated with 2 μg /mL of secondary FITC-conjugated goat anti-human IgG (Novus Biologicals, Centennial CO USA) for 1 hr at room temperature in the dark, then washed a final time with PBS and resuspended in 2% paraformaldehyde.

Techniques: Binding Assay, Immunofluorescence, Flow Cytometry, Fluorescence, Incubation, Concentration Assay

Journal: Cellular and Molecular Gastroenterology and Hepatology

Article Title: Unique Neural Circuit Connectivity of Mouse Proximal, Middle, and Distal Colon Defines Regional Colonic Motor Patterns

doi: 10.1016/j.jcmgh.2021.08.016

Figure Lengend Snippet: Primary and Secondary Antibodies

Article Snippet: Human (IgG, H+L) , Goat , DyLight 405 , 1:500 , Novus Biologicals (NBP1-72871) lLittleton, CO , RRID: AB_11008096.

Techniques:

Journal: Advanced healthcare materials

Article Title: Architectural Modification of Conformal PEG-Bottlebrush Coatings Minimizes Anti-PEG Antigenicity While Preserving Stealth Properties

doi: 10.1002/adhm.201801177

Figure Lengend Snippet: Direct comparison of reactivity of APAs toward POEGMA brushes with different EG sidechain lengths versus linear PEG (MW = 20 K). a, Schematic of printing microspots of PEG20K-BSA Ag onto a background of POEGMA brush surfaces. Surfaces were incubated with a dilution series of rabbit-derived pAPA1 in serum, labeled with Cy5-anti-rabbit dAb, and then read by a fluorescence scanner. b, Spatial intensity plots of Cy5 fluorescence from EG2-OMe polymer brush surfaces functionalized by PEG20K-BSA Ag microspots (outlined by white dashes). Shown are 330 × 330 μm regions corresponding to surfaces (containing a single Ag microspot) exposed to serum alone (left) versus serum spiked with 2 μg/mL pAPA1 (right). c, Concentration curves of pAPA1 binding measured by fluorescence intensity from PEG20K-BSA Ag microspots (black squares) versus that from EG2-OMe POEGMA background (open circles). Similar spatial intensity plots and concentration curves as shown for EG2-OMe in (b, c) are shown for EG3-OMe in (d, e) and EG5-OMe in (f, g). Data plotted in (c, e, g) represent mean ± s.d. (n = 3). LODs determined from PEG20K-BSA microspots versus polymer background (LODAg vs. LODbkg, respectively) are displayed adjacent to each curve.

Article Snippet: Next, the surfaces were exposed to a 1 μg/mL solution of Cy5-goat-anti-human IgG detection Ab (R & D Systems, Inc.) for 30 min.

Techniques: Comparison, Incubation, Derivative Assay, Labeling, Fluorescence, Polymer, Concentration Assay, Binding Assay

Journal: Advanced healthcare materials

Article Title: Architectural Modification of Conformal PEG-Bottlebrush Coatings Minimizes Anti-PEG Antigenicity While Preserving Stealth Properties

doi: 10.1002/adhm.201801177

Figure Lengend Snippet: Screening POEGMA bottlebrush surfaces for immune reactivity toward a polyclonal APA (pAPA1). a, Schematic of pAPA1 fluoroimmunoassay. Surfaces were incubated with a solution of rabbit-derived pAPA1 spiked into undiluted calf serum, rinsed, and then labeled with Cy5-donkey-α-rabbit dAbs, and then imaged with a fluorescence scanner. b, c, Representative Cy5 channel fluorescence images (b) and quantitation of mean fluorescence intensities (c). Results in (c) are plotted as mean ± 95% CI for n ≥ 4 replicates. Vehicle-only and rabbit-IgG (not specifically reactive to PEG) controls are also included to show baseline values. Incubating surfaces with pAPA1 leads to significant fluorescence, indicating APA binding, from EG5-OMe, EG6-OH, and EG9-OMe surfaces, but not from bare, EG1-OMe, EG2-OM2, or EG3-OMe surfaces. Bars marked with different letters indicate significant differences within the pAPA1-treated groups by multiple comparison testing in one-way ANOVA (Tukey post hoc test, p ≤ 0.05).

Article Snippet: Next, the surfaces were exposed to a 1 μg/mL solution of Cy5-goat-anti-human IgG detection Ab (R & D Systems, Inc.) for 30 min.

Techniques: Incubation, Derivative Assay, Labeling, Fluorescence, Quantitation Assay, Binding Assay, Comparison

Journal: Advanced healthcare materials

Article Title: Architectural Modification of Conformal PEG-Bottlebrush Coatings Minimizes Anti-PEG Antigenicity While Preserving Stealth Properties

doi: 10.1002/adhm.201801177

Figure Lengend Snippet: Screening POEGMA brush surfaces for protein adsorption and cell adhesion. a, Schematic of surface fluorescence assay used to evaluate protein fouling. Cy5-labeled BSA was incubated on surfaces, rinsed and then read with a scanner for residual fluorescence. b, c, Representative Cy5 channel fluorescence images (b) and quantitation of mean ± 95% CI fluorescence intensities (c) (n = 3 for bare glass, and n ≥ 6 for others). Vehicle groups are plotted for comparison. Bars marked with different letters indicate significant differences within the Cy5-BSA-treated groups by multiple comparison testing in one-way ANOVA (Tukey post hoc test, p ≤ 0.05). d, Schematic of in vitro cell adhesion assay. NIH 3T3 cells expressing GFP (3T3-GFP) were incubated on surfaces in complete medium, washed, and then imaged for residual fluorescence on GFP channel by epifluorescence imaging. e, f, Representative epifluorescence images of cells (e) and quantitation of cell adhesion to surfaces (f) expressed as mean %FOV ± 95% CI (n = 6). Bars marked with different letters indicate significantly different groups by multiple comparison testing in one-way ANOVA (Tukey post hoc test, p ≤ 0.05)

Article Snippet: Next, the surfaces were exposed to a 1 μg/mL solution of Cy5-goat-anti-human IgG detection Ab (R & D Systems, Inc.) for 30 min.

Techniques: Adsorption, Fluorescence, Labeling, Incubation, Quantitation Assay, Comparison, In Vitro, Cell Adhesion Assay, Expressing, Imaging

Journal: Advanced healthcare materials

Article Title: Architectural Modification of Conformal PEG-Bottlebrush Coatings Minimizes Anti-PEG Antigenicity While Preserving Stealth Properties

doi: 10.1002/adhm.201801177

Figure Lengend Snippet: Evaluating interference from APA reactivity in indirect sandwich immunoassays (ISIAs) for antibody detection (“serology”) fabricated on polymer bottlebrushes. a, Schematic of serological antibody ISIA. ISIAs comprised of p24 Ag spotted onto bottlebrush overlayers were incubated with a dilution series of rabbit anti-HIV p24 polyclonal Ab, either with (bottom pathway) or without (top pathway) the presence of APA interferent (pAPA1). Surfaces were labeled with Cy5-donkey-anti-rabbit dAb, and then read by a scanner. b, c, d, e, f, Concentration binding curves for detecting polyclonal anti-p24 Ab (analyte) on polymer brush-based ISIAs, either with or without 100 ng/mL of APA interferent (black squares and open circles, respectively). LODs for each curve are provided in ng/mL, except for the EG5-OMe curve run with APA interferent, which was not calculated due to high background noise. Each data point represents mean ± s.d. from duplicate runs.

Article Snippet: Next, the surfaces were exposed to a 1 μg/mL solution of Cy5-goat-anti-human IgG detection Ab (R & D Systems, Inc.) for 30 min.

Techniques: Polymer, Incubation, Labeling, Concentration Assay, Binding Assay

Journal: Advanced healthcare materials

Article Title: Architectural Modification of Conformal PEG-Bottlebrush Coatings Minimizes Anti-PEG Antigenicity While Preserving Stealth Properties

doi: 10.1002/adhm.201801177

Figure Lengend Snippet: Reactivity of backbone-selective versus endgroup selective APAs toward EG2-OMe, EG3-OMe, and EG5-OMe POEGMA brushes. a, Schematic of backbone-selective (blue) versus endgroup-selective (tan) APA binding to PEG backbone and methoxy terminus of a bottlebrush, respectively. b, Schematic of surface fluoroimmunoassay for APA binding. Surfaces were incubated with a solution of APA-spiked calf serum, then labeled with Cy5-conjugated dAbs, and then read with a scanner. c–f, Reactivity of polyclonal APAs toward bottlebrush surfaces with known selectivity for PEG endgroups (pAPA1) versus backbone (pAPA2) (c, d), and similar plots shown for endgroup-selective (e-mAPA) versus backbone-selective (b-mAPA) monoclonal APAs (e, f) as assessed by surface fluoroimmunoassays. Data are plotted as mean fluorescence intensities ± s.d. (n = 9). Bars marked with different letters indicate significant differences by multiple comparison testing in one-way ANOVA (Tukey post hoc test, p ≤ 0.05).

Article Snippet: Next, the surfaces were exposed to a 1 μg/mL solution of Cy5-goat-anti-human IgG detection Ab (R & D Systems, Inc.) for 30 min.

Techniques: Binding Assay, Incubation, Labeling, Fluorescence, Comparison