recombinant human cd276 extracellular domain  (Sino Biological)


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    Name:
    B7 H3 Protein Human Recombinant Biotinylated
    Description:
    A DNA sequence encoding the human CD276 NP 001019907 1 Met1 Thr461 was expressed with the Fc region of human IgG1 at the C terminus The purified protein was biotinylated in vitro
    Catalog Number:
    11188-h02h-b
    Product Aliases:
    4Ig-B7-H3 Protein Human, B7-H3 Protein Human, B7H3 Protein Human, B7RP-2 Protein Human
    Price:
    None
    Host:
    HEK293 Cells
    Category:
    recombinant protein
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    Structured Review

    Sino Biological recombinant human cd276 extracellular domain
    Characterization of parental and evolved <t>CD276-binding</t> Affibodies. (A) affibody variants AC2, AC9, AC12, and AC16 were characterized for their binding affinity (K D ). Blue lettering indicates diversified residues in the helix-walking libraries. (B) Purified affibody variants AC2 (upper-left), AC9 (upper-right), AC12 (lower-left), and AC16 (lower-right) were used to label MS1-CD276 cells at the indicated concentrations. Binding was quantified by flow cytometry. The best-fit estimate of K D and 68% confidence interval are indicated by solid and dashed lines, respectively. (C) Purified affibody AC12 was analyzed by circular dichroism spectroscopy in triplicate between 200 and 260 nm wavelengths before (solid) and after (dashed) thermal denaturation and cooling. (D) Purified affibody AC12 was scanned at a wavelength of 220 nm during heating from 20 to 98 °C (1 °C/min). The midpoint of thermal denaturation (T m ) was calculated by linear least-squares regression using a two-state protein unfolding model.
    A DNA sequence encoding the human CD276 NP 001019907 1 Met1 Thr461 was expressed with the Fc region of human IgG1 at the C terminus The purified protein was biotinylated in vitro
    https://www.bioz.com/result/recombinant human cd276 extracellular domain/product/Sino Biological
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    recombinant human cd276 extracellular domain - by Bioz Stars, 2021-02
    94/100 stars

    Images

    1) Product Images from "Cellular-Based Selections Aid Yeast-Display Discovery of Genuine Cell-Binding Ligands: Targeting Oncology Vascular Biomarker CD276"

    Article Title: Cellular-Based Selections Aid Yeast-Display Discovery of Genuine Cell-Binding Ligands: Targeting Oncology Vascular Biomarker CD276

    Journal: ACS combinatorial science

    doi: 10.1021/acscombsci.8b00156

    Characterization of parental and evolved CD276-binding Affibodies. (A) affibody variants AC2, AC9, AC12, and AC16 were characterized for their binding affinity (K D ). Blue lettering indicates diversified residues in the helix-walking libraries. (B) Purified affibody variants AC2 (upper-left), AC9 (upper-right), AC12 (lower-left), and AC16 (lower-right) were used to label MS1-CD276 cells at the indicated concentrations. Binding was quantified by flow cytometry. The best-fit estimate of K D and 68% confidence interval are indicated by solid and dashed lines, respectively. (C) Purified affibody AC12 was analyzed by circular dichroism spectroscopy in triplicate between 200 and 260 nm wavelengths before (solid) and after (dashed) thermal denaturation and cooling. (D) Purified affibody AC12 was scanned at a wavelength of 220 nm during heating from 20 to 98 °C (1 °C/min). The midpoint of thermal denaturation (T m ) was calculated by linear least-squares regression using a two-state protein unfolding model.
    Figure Legend Snippet: Characterization of parental and evolved CD276-binding Affibodies. (A) affibody variants AC2, AC9, AC12, and AC16 were characterized for their binding affinity (K D ). Blue lettering indicates diversified residues in the helix-walking libraries. (B) Purified affibody variants AC2 (upper-left), AC9 (upper-right), AC12 (lower-left), and AC16 (lower-right) were used to label MS1-CD276 cells at the indicated concentrations. Binding was quantified by flow cytometry. The best-fit estimate of K D and 68% confidence interval are indicated by solid and dashed lines, respectively. (C) Purified affibody AC12 was analyzed by circular dichroism spectroscopy in triplicate between 200 and 260 nm wavelengths before (solid) and after (dashed) thermal denaturation and cooling. (D) Purified affibody AC12 was scanned at a wavelength of 220 nm during heating from 20 to 98 °C (1 °C/min). The midpoint of thermal denaturation (T m ) was calculated by linear least-squares regression using a two-state protein unfolding model.

    Techniques Used: Binding Assay, Purification, Flow Cytometry, Spectroscopy

    Ligand discovery methods. An affibody library and a fibronectin domain library were sorted for ligands that bound CD276 or Thy1 specifically. Libraries were sorted by five different schemes: 1) magnetic bead selection with recombinant extracellular domains followed by FACS with recombinant extracellular domains, 2) magnetic bead selection followed by FACS with detergent solubilized cell lysate, 3) magnetic bead selection followed by cell panning selection, 4) cell panning selection with magnetic bead depletion, and 5) cell panning selection.
    Figure Legend Snippet: Ligand discovery methods. An affibody library and a fibronectin domain library were sorted for ligands that bound CD276 or Thy1 specifically. Libraries were sorted by five different schemes: 1) magnetic bead selection with recombinant extracellular domains followed by FACS with recombinant extracellular domains, 2) magnetic bead selection followed by FACS with detergent solubilized cell lysate, 3) magnetic bead selection followed by cell panning selection, 4) cell panning selection with magnetic bead depletion, and 5) cell panning selection.

    Techniques Used: Selection, Recombinant, FACS

    Optimization of incubation time for yeast-displayed ligand enrichment. Yeast displaying affibody clones LS (A) or HS (B) were panned for binding to adherent MS1-CD276 with varying incubation times. Recoveries are presented as the mean ± error deviation of 7–12 trials.
    Figure Legend Snippet: Optimization of incubation time for yeast-displayed ligand enrichment. Yeast displaying affibody clones LS (A) or HS (B) were panned for binding to adherent MS1-CD276 with varying incubation times. Recoveries are presented as the mean ± error deviation of 7–12 trials.

    Techniques Used: Incubation, Clone Assay, Binding Assay

    Enriched ligands evaluated for cellular binding. Yeast-displayed ligand populations were enriched through sequential rounds of selection for binding against CD276 and Thy1 using cell panning methods. Rec. + Panning indicates recombinant target-coated magnetic bead selections followed by cellular panning. Depleted Panning and Panning indicate cellular panning with or without, respectively, depletion of non-specific binders via streptavidin-coated magnetic beads. Binding specificity for each enriched population was assessed by cell panning. Yeast were panned for binders on monolayers of target-positive MS1 cells (blue) or target-negative MS1 cells as a negative control (orange). In the depleted panning case, recovery of yeast from the first (white) and second (gray) magnetic beads was also quantified. Data represent single-run analyses during the course of each discovery campaign.
    Figure Legend Snippet: Enriched ligands evaluated for cellular binding. Yeast-displayed ligand populations were enriched through sequential rounds of selection for binding against CD276 and Thy1 using cell panning methods. Rec. + Panning indicates recombinant target-coated magnetic bead selections followed by cellular panning. Depleted Panning and Panning indicate cellular panning with or without, respectively, depletion of non-specific binders via streptavidin-coated magnetic beads. Binding specificity for each enriched population was assessed by cell panning. Yeast were panned for binders on monolayers of target-positive MS1 cells (blue) or target-negative MS1 cells as a negative control (orange). In the depleted panning case, recovery of yeast from the first (white) and second (gray) magnetic beads was also quantified. Data represent single-run analyses during the course of each discovery campaign.

    Techniques Used: Binding Assay, Selection, Recombinant, Magnetic Beads, Negative Control

    Enriched ligands evaluated for soluble extracellular domain and detergent-solubilized cell lysate binding. Yeast-displayed ligand populations were enriched for binders against CD276 and Thy1 using soluble extracellular domains immobilized on magnetic beads followed by FACS with either soluble extracellular domains or detergent-solubilized cell lysates. Binding specificity for each enriched population was assessed by comparison to negative controls. For FACS with soluble extracellular domains, yeast were labeled with 100 nM soluble CD276 or Thy1-Fc extracellular domains (blue) or irrelevant negative control proteins (300 nM streptavidin for CD276 or 100 nM human IgG for Thy1-Fc; orange) (A-D). For selection of ligands with detergent-solubilized cell lysates against CD276, yeast were labeled with MS1-CD276 lysate (blue) or MS1-Thy1 lysate (orange). For selection of ligands with detergent-solubilized cell lysates against Thy1, yeast were labeled with MS1-Thy1 lysate (blue) or MS1-CD276 lysate (orange) (E-H).
    Figure Legend Snippet: Enriched ligands evaluated for soluble extracellular domain and detergent-solubilized cell lysate binding. Yeast-displayed ligand populations were enriched for binders against CD276 and Thy1 using soluble extracellular domains immobilized on magnetic beads followed by FACS with either soluble extracellular domains or detergent-solubilized cell lysates. Binding specificity for each enriched population was assessed by comparison to negative controls. For FACS with soluble extracellular domains, yeast were labeled with 100 nM soluble CD276 or Thy1-Fc extracellular domains (blue) or irrelevant negative control proteins (300 nM streptavidin for CD276 or 100 nM human IgG for Thy1-Fc; orange) (A-D). For selection of ligands with detergent-solubilized cell lysates against CD276, yeast were labeled with MS1-CD276 lysate (blue) or MS1-Thy1 lysate (orange). For selection of ligands with detergent-solubilized cell lysates against Thy1, yeast were labeled with MS1-Thy1 lysate (blue) or MS1-CD276 lysate (orange) (E-H).

    Techniques Used: Binding Assay, Magnetic Beads, FACS, Labeling, Negative Control, Selection

    Depletion of non-specific binders with mammalian cell pre-blocking. Mixtures of high-yield (A) or low-yield (B) CD276-specific, non-specific, and non-binding yeast were incubated with either CD276-negative disadhered MS1-Thy1 cells (mammalian cell pre-block) or buffer only (no depletion) followed by incubation with MS1-CD276 cell monolayers. Enrichment ratios for CD276-specific (black), non-specific (gray), or non-binding (white) clones are shown as the mean ± standard deviation of 6–12 trials. (C). Specific (black) or non-target-specific (grey) clones were subjected to selection with or without mammalian cell pre-blocking for a total of 13 clone. Each point represents the yield of a single well of selection using pre-blocking normalized by a corresponding well selected without pre-blocking. Each clone was panned either in duplicate or triplicate.
    Figure Legend Snippet: Depletion of non-specific binders with mammalian cell pre-blocking. Mixtures of high-yield (A) or low-yield (B) CD276-specific, non-specific, and non-binding yeast were incubated with either CD276-negative disadhered MS1-Thy1 cells (mammalian cell pre-block) or buffer only (no depletion) followed by incubation with MS1-CD276 cell monolayers. Enrichment ratios for CD276-specific (black), non-specific (gray), or non-binding (white) clones are shown as the mean ± standard deviation of 6–12 trials. (C). Specific (black) or non-target-specific (grey) clones were subjected to selection with or without mammalian cell pre-blocking for a total of 13 clone. Each point represents the yield of a single well of selection using pre-blocking normalized by a corresponding well selected without pre-blocking. Each clone was panned either in duplicate or triplicate.

    Techniques Used: Blocking Assay, Binding Assay, Incubation, Standard Deviation, Selection

    Sequential depletion of non-specific binders with mammalian cell monolayers. Mixtures of high-yield (A) or low-yield (B) CD276-specific, non-specific, and non-binding yeast were subjected to selection with 0, 2, 4, or 6 depletion steps followed by a single enrichment step. All individual enrichment ratios are shown for 3–10 trials as well as the average.
    Figure Legend Snippet: Sequential depletion of non-specific binders with mammalian cell monolayers. Mixtures of high-yield (A) or low-yield (B) CD276-specific, non-specific, and non-binding yeast were subjected to selection with 0, 2, 4, or 6 depletion steps followed by a single enrichment step. All individual enrichment ratios are shown for 3–10 trials as well as the average.

    Techniques Used: Binding Assay, Selection

    Evolved populations of yeast-displayed ligands show increasing binding to CD276 lysate while maintaining target specificity. (A) Yeast populations collected after preliminary sorting on recombinant target beads (first column) or final sorting on target-positive lysate (second column) were labelled with 150 nM target cell lysate and analyzed for binding by flow cytometry. Yeast collected after sorting on target-positive lysate (third column) were labelled with 150 nM target-negative cell lysate and analyzed for specificity by flow cytometry. S ubstantial binding improvement can be observed between preliminary sorting and the final population, with low cross-reactivity to target-negative lysate. (B) A mixture of the merged library and triple-sorted single-helix library was labelled with 0.5 nM target lysate (left) or 50 nM target-negative lysate (right) and analyzed for binding by flow cytometry. The population shows significantly binding to target-positive lysate with minimal cross-reactivity to target-negative lysate.
    Figure Legend Snippet: Evolved populations of yeast-displayed ligands show increasing binding to CD276 lysate while maintaining target specificity. (A) Yeast populations collected after preliminary sorting on recombinant target beads (first column) or final sorting on target-positive lysate (second column) were labelled with 150 nM target cell lysate and analyzed for binding by flow cytometry. Yeast collected after sorting on target-positive lysate (third column) were labelled with 150 nM target-negative cell lysate and analyzed for specificity by flow cytometry. S ubstantial binding improvement can be observed between preliminary sorting and the final population, with low cross-reactivity to target-negative lysate. (B) A mixture of the merged library and triple-sorted single-helix library was labelled with 0.5 nM target lysate (left) or 50 nM target-negative lysate (right) and analyzed for binding by flow cytometry. The population shows significantly binding to target-positive lysate with minimal cross-reactivity to target-negative lysate.

    Techniques Used: Binding Assay, Recombinant, Flow Cytometry

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  • 94
    Sino Biological recombinant human cd276 extracellular domain
    Characterization of parental and evolved <t>CD276-binding</t> Affibodies. (A) affibody variants AC2, AC9, AC12, and AC16 were characterized for their binding affinity (K D ). Blue lettering indicates diversified residues in the helix-walking libraries. (B) Purified affibody variants AC2 (upper-left), AC9 (upper-right), AC12 (lower-left), and AC16 (lower-right) were used to label MS1-CD276 cells at the indicated concentrations. Binding was quantified by flow cytometry. The best-fit estimate of K D and 68% confidence interval are indicated by solid and dashed lines, respectively. (C) Purified affibody AC12 was analyzed by circular dichroism spectroscopy in triplicate between 200 and 260 nm wavelengths before (solid) and after (dashed) thermal denaturation and cooling. (D) Purified affibody AC12 was scanned at a wavelength of 220 nm during heating from 20 to 98 °C (1 °C/min). The midpoint of thermal denaturation (T m ) was calculated by linear least-squares regression using a two-state protein unfolding model.
    Recombinant Human Cd276 Extracellular Domain, supplied by Sino Biological, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/recombinant human cd276 extracellular domain/product/Sino Biological
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    recombinant human cd276 extracellular domain - by Bioz Stars, 2021-02
    94/100 stars
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    N/A
    Produced in rabbits immunized with purified recombinant Human REG4 rh REG4 Catalog 11186 H08H NP 001152824 1 Met 1 Pro 158 REG4 specific IgG was purified by human REG4 affinity
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    N/A
    A DNA sequence encoding the human CD276 Q5ZPR3 1 extracellular domain Met 1 Thr 461 was expressed with a polyhistidine tag at the C terminus
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    Image Search Results


    Characterization of parental and evolved CD276-binding Affibodies. (A) affibody variants AC2, AC9, AC12, and AC16 were characterized for their binding affinity (K D ). Blue lettering indicates diversified residues in the helix-walking libraries. (B) Purified affibody variants AC2 (upper-left), AC9 (upper-right), AC12 (lower-left), and AC16 (lower-right) were used to label MS1-CD276 cells at the indicated concentrations. Binding was quantified by flow cytometry. The best-fit estimate of K D and 68% confidence interval are indicated by solid and dashed lines, respectively. (C) Purified affibody AC12 was analyzed by circular dichroism spectroscopy in triplicate between 200 and 260 nm wavelengths before (solid) and after (dashed) thermal denaturation and cooling. (D) Purified affibody AC12 was scanned at a wavelength of 220 nm during heating from 20 to 98 °C (1 °C/min). The midpoint of thermal denaturation (T m ) was calculated by linear least-squares regression using a two-state protein unfolding model.

    Journal: ACS combinatorial science

    Article Title: Cellular-Based Selections Aid Yeast-Display Discovery of Genuine Cell-Binding Ligands: Targeting Oncology Vascular Biomarker CD276

    doi: 10.1021/acscombsci.8b00156

    Figure Lengend Snippet: Characterization of parental and evolved CD276-binding Affibodies. (A) affibody variants AC2, AC9, AC12, and AC16 were characterized for their binding affinity (K D ). Blue lettering indicates diversified residues in the helix-walking libraries. (B) Purified affibody variants AC2 (upper-left), AC9 (upper-right), AC12 (lower-left), and AC16 (lower-right) were used to label MS1-CD276 cells at the indicated concentrations. Binding was quantified by flow cytometry. The best-fit estimate of K D and 68% confidence interval are indicated by solid and dashed lines, respectively. (C) Purified affibody AC12 was analyzed by circular dichroism spectroscopy in triplicate between 200 and 260 nm wavelengths before (solid) and after (dashed) thermal denaturation and cooling. (D) Purified affibody AC12 was scanned at a wavelength of 220 nm during heating from 20 to 98 °C (1 °C/min). The midpoint of thermal denaturation (T m ) was calculated by linear least-squares regression using a two-state protein unfolding model.

    Article Snippet: Recombinant human CD276 extracellular domain (Sino Biological, Cat: 11188-H08H-B) was obtained already biotinylated from the manufacturer.

    Techniques: Binding Assay, Purification, Flow Cytometry, Spectroscopy

    Ligand discovery methods. An affibody library and a fibronectin domain library were sorted for ligands that bound CD276 or Thy1 specifically. Libraries were sorted by five different schemes: 1) magnetic bead selection with recombinant extracellular domains followed by FACS with recombinant extracellular domains, 2) magnetic bead selection followed by FACS with detergent solubilized cell lysate, 3) magnetic bead selection followed by cell panning selection, 4) cell panning selection with magnetic bead depletion, and 5) cell panning selection.

    Journal: ACS combinatorial science

    Article Title: Cellular-Based Selections Aid Yeast-Display Discovery of Genuine Cell-Binding Ligands: Targeting Oncology Vascular Biomarker CD276

    doi: 10.1021/acscombsci.8b00156

    Figure Lengend Snippet: Ligand discovery methods. An affibody library and a fibronectin domain library were sorted for ligands that bound CD276 or Thy1 specifically. Libraries were sorted by five different schemes: 1) magnetic bead selection with recombinant extracellular domains followed by FACS with recombinant extracellular domains, 2) magnetic bead selection followed by FACS with detergent solubilized cell lysate, 3) magnetic bead selection followed by cell panning selection, 4) cell panning selection with magnetic bead depletion, and 5) cell panning selection.

    Article Snippet: Recombinant human CD276 extracellular domain (Sino Biological, Cat: 11188-H08H-B) was obtained already biotinylated from the manufacturer.

    Techniques: Selection, Recombinant, FACS

    Optimization of incubation time for yeast-displayed ligand enrichment. Yeast displaying affibody clones LS (A) or HS (B) were panned for binding to adherent MS1-CD276 with varying incubation times. Recoveries are presented as the mean ± error deviation of 7–12 trials.

    Journal: ACS combinatorial science

    Article Title: Cellular-Based Selections Aid Yeast-Display Discovery of Genuine Cell-Binding Ligands: Targeting Oncology Vascular Biomarker CD276

    doi: 10.1021/acscombsci.8b00156

    Figure Lengend Snippet: Optimization of incubation time for yeast-displayed ligand enrichment. Yeast displaying affibody clones LS (A) or HS (B) were panned for binding to adherent MS1-CD276 with varying incubation times. Recoveries are presented as the mean ± error deviation of 7–12 trials.

    Article Snippet: Recombinant human CD276 extracellular domain (Sino Biological, Cat: 11188-H08H-B) was obtained already biotinylated from the manufacturer.

    Techniques: Incubation, Clone Assay, Binding Assay

    Enriched ligands evaluated for cellular binding. Yeast-displayed ligand populations were enriched through sequential rounds of selection for binding against CD276 and Thy1 using cell panning methods. Rec. + Panning indicates recombinant target-coated magnetic bead selections followed by cellular panning. Depleted Panning and Panning indicate cellular panning with or without, respectively, depletion of non-specific binders via streptavidin-coated magnetic beads. Binding specificity for each enriched population was assessed by cell panning. Yeast were panned for binders on monolayers of target-positive MS1 cells (blue) or target-negative MS1 cells as a negative control (orange). In the depleted panning case, recovery of yeast from the first (white) and second (gray) magnetic beads was also quantified. Data represent single-run analyses during the course of each discovery campaign.

    Journal: ACS combinatorial science

    Article Title: Cellular-Based Selections Aid Yeast-Display Discovery of Genuine Cell-Binding Ligands: Targeting Oncology Vascular Biomarker CD276

    doi: 10.1021/acscombsci.8b00156

    Figure Lengend Snippet: Enriched ligands evaluated for cellular binding. Yeast-displayed ligand populations were enriched through sequential rounds of selection for binding against CD276 and Thy1 using cell panning methods. Rec. + Panning indicates recombinant target-coated magnetic bead selections followed by cellular panning. Depleted Panning and Panning indicate cellular panning with or without, respectively, depletion of non-specific binders via streptavidin-coated magnetic beads. Binding specificity for each enriched population was assessed by cell panning. Yeast were panned for binders on monolayers of target-positive MS1 cells (blue) or target-negative MS1 cells as a negative control (orange). In the depleted panning case, recovery of yeast from the first (white) and second (gray) magnetic beads was also quantified. Data represent single-run analyses during the course of each discovery campaign.

    Article Snippet: Recombinant human CD276 extracellular domain (Sino Biological, Cat: 11188-H08H-B) was obtained already biotinylated from the manufacturer.

    Techniques: Binding Assay, Selection, Recombinant, Magnetic Beads, Negative Control

    Enriched ligands evaluated for soluble extracellular domain and detergent-solubilized cell lysate binding. Yeast-displayed ligand populations were enriched for binders against CD276 and Thy1 using soluble extracellular domains immobilized on magnetic beads followed by FACS with either soluble extracellular domains or detergent-solubilized cell lysates. Binding specificity for each enriched population was assessed by comparison to negative controls. For FACS with soluble extracellular domains, yeast were labeled with 100 nM soluble CD276 or Thy1-Fc extracellular domains (blue) or irrelevant negative control proteins (300 nM streptavidin for CD276 or 100 nM human IgG for Thy1-Fc; orange) (A-D). For selection of ligands with detergent-solubilized cell lysates against CD276, yeast were labeled with MS1-CD276 lysate (blue) or MS1-Thy1 lysate (orange). For selection of ligands with detergent-solubilized cell lysates against Thy1, yeast were labeled with MS1-Thy1 lysate (blue) or MS1-CD276 lysate (orange) (E-H).

    Journal: ACS combinatorial science

    Article Title: Cellular-Based Selections Aid Yeast-Display Discovery of Genuine Cell-Binding Ligands: Targeting Oncology Vascular Biomarker CD276

    doi: 10.1021/acscombsci.8b00156

    Figure Lengend Snippet: Enriched ligands evaluated for soluble extracellular domain and detergent-solubilized cell lysate binding. Yeast-displayed ligand populations were enriched for binders against CD276 and Thy1 using soluble extracellular domains immobilized on magnetic beads followed by FACS with either soluble extracellular domains or detergent-solubilized cell lysates. Binding specificity for each enriched population was assessed by comparison to negative controls. For FACS with soluble extracellular domains, yeast were labeled with 100 nM soluble CD276 or Thy1-Fc extracellular domains (blue) or irrelevant negative control proteins (300 nM streptavidin for CD276 or 100 nM human IgG for Thy1-Fc; orange) (A-D). For selection of ligands with detergent-solubilized cell lysates against CD276, yeast were labeled with MS1-CD276 lysate (blue) or MS1-Thy1 lysate (orange). For selection of ligands with detergent-solubilized cell lysates against Thy1, yeast were labeled with MS1-Thy1 lysate (blue) or MS1-CD276 lysate (orange) (E-H).

    Article Snippet: Recombinant human CD276 extracellular domain (Sino Biological, Cat: 11188-H08H-B) was obtained already biotinylated from the manufacturer.

    Techniques: Binding Assay, Magnetic Beads, FACS, Labeling, Negative Control, Selection

    Depletion of non-specific binders with mammalian cell pre-blocking. Mixtures of high-yield (A) or low-yield (B) CD276-specific, non-specific, and non-binding yeast were incubated with either CD276-negative disadhered MS1-Thy1 cells (mammalian cell pre-block) or buffer only (no depletion) followed by incubation with MS1-CD276 cell monolayers. Enrichment ratios for CD276-specific (black), non-specific (gray), or non-binding (white) clones are shown as the mean ± standard deviation of 6–12 trials. (C). Specific (black) or non-target-specific (grey) clones were subjected to selection with or without mammalian cell pre-blocking for a total of 13 clone. Each point represents the yield of a single well of selection using pre-blocking normalized by a corresponding well selected without pre-blocking. Each clone was panned either in duplicate or triplicate.

    Journal: ACS combinatorial science

    Article Title: Cellular-Based Selections Aid Yeast-Display Discovery of Genuine Cell-Binding Ligands: Targeting Oncology Vascular Biomarker CD276

    doi: 10.1021/acscombsci.8b00156

    Figure Lengend Snippet: Depletion of non-specific binders with mammalian cell pre-blocking. Mixtures of high-yield (A) or low-yield (B) CD276-specific, non-specific, and non-binding yeast were incubated with either CD276-negative disadhered MS1-Thy1 cells (mammalian cell pre-block) or buffer only (no depletion) followed by incubation with MS1-CD276 cell monolayers. Enrichment ratios for CD276-specific (black), non-specific (gray), or non-binding (white) clones are shown as the mean ± standard deviation of 6–12 trials. (C). Specific (black) or non-target-specific (grey) clones were subjected to selection with or without mammalian cell pre-blocking for a total of 13 clone. Each point represents the yield of a single well of selection using pre-blocking normalized by a corresponding well selected without pre-blocking. Each clone was panned either in duplicate or triplicate.

    Article Snippet: Recombinant human CD276 extracellular domain (Sino Biological, Cat: 11188-H08H-B) was obtained already biotinylated from the manufacturer.

    Techniques: Blocking Assay, Binding Assay, Incubation, Standard Deviation, Selection

    Sequential depletion of non-specific binders with mammalian cell monolayers. Mixtures of high-yield (A) or low-yield (B) CD276-specific, non-specific, and non-binding yeast were subjected to selection with 0, 2, 4, or 6 depletion steps followed by a single enrichment step. All individual enrichment ratios are shown for 3–10 trials as well as the average.

    Journal: ACS combinatorial science

    Article Title: Cellular-Based Selections Aid Yeast-Display Discovery of Genuine Cell-Binding Ligands: Targeting Oncology Vascular Biomarker CD276

    doi: 10.1021/acscombsci.8b00156

    Figure Lengend Snippet: Sequential depletion of non-specific binders with mammalian cell monolayers. Mixtures of high-yield (A) or low-yield (B) CD276-specific, non-specific, and non-binding yeast were subjected to selection with 0, 2, 4, or 6 depletion steps followed by a single enrichment step. All individual enrichment ratios are shown for 3–10 trials as well as the average.

    Article Snippet: Recombinant human CD276 extracellular domain (Sino Biological, Cat: 11188-H08H-B) was obtained already biotinylated from the manufacturer.

    Techniques: Binding Assay, Selection

    Evolved populations of yeast-displayed ligands show increasing binding to CD276 lysate while maintaining target specificity. (A) Yeast populations collected after preliminary sorting on recombinant target beads (first column) or final sorting on target-positive lysate (second column) were labelled with 150 nM target cell lysate and analyzed for binding by flow cytometry. Yeast collected after sorting on target-positive lysate (third column) were labelled with 150 nM target-negative cell lysate and analyzed for specificity by flow cytometry. S ubstantial binding improvement can be observed between preliminary sorting and the final population, with low cross-reactivity to target-negative lysate. (B) A mixture of the merged library and triple-sorted single-helix library was labelled with 0.5 nM target lysate (left) or 50 nM target-negative lysate (right) and analyzed for binding by flow cytometry. The population shows significantly binding to target-positive lysate with minimal cross-reactivity to target-negative lysate.

    Journal: ACS combinatorial science

    Article Title: Cellular-Based Selections Aid Yeast-Display Discovery of Genuine Cell-Binding Ligands: Targeting Oncology Vascular Biomarker CD276

    doi: 10.1021/acscombsci.8b00156

    Figure Lengend Snippet: Evolved populations of yeast-displayed ligands show increasing binding to CD276 lysate while maintaining target specificity. (A) Yeast populations collected after preliminary sorting on recombinant target beads (first column) or final sorting on target-positive lysate (second column) were labelled with 150 nM target cell lysate and analyzed for binding by flow cytometry. Yeast collected after sorting on target-positive lysate (third column) were labelled with 150 nM target-negative cell lysate and analyzed for specificity by flow cytometry. S ubstantial binding improvement can be observed between preliminary sorting and the final population, with low cross-reactivity to target-negative lysate. (B) A mixture of the merged library and triple-sorted single-helix library was labelled with 0.5 nM target lysate (left) or 50 nM target-negative lysate (right) and analyzed for binding by flow cytometry. The population shows significantly binding to target-positive lysate with minimal cross-reactivity to target-negative lysate.

    Article Snippet: Recombinant human CD276 extracellular domain (Sino Biological, Cat: 11188-H08H-B) was obtained already biotinylated from the manufacturer.

    Techniques: Binding Assay, Recombinant, Flow Cytometry