BioChip Ventures Division

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a 3-Dimensional Microarray Substrate. BioChip Ventures Division. What is a microarray?. Advantages multiplexing and miniaturization  throughput parallel analysis sample volume reduction. target. probe. Protein Microarray Applications. * DNA - protein interaction - PowerPoint PPT Presentation

Text of BioChip Ventures Division

  • BioChip Ventures Division

    a 3-Dimensional Microarray Substrate

  • What is a microarray?probetargetAdvantages multiplexing and miniaturization throughput parallel analysis sample volume reduction

  • Protein Microarray Applications

    * DNA - protein interaction

    * Protein - protein interactions

    * Enzyme-substrate analysis

    * Protein profiling

    * Antibody characterization

    * Small molecule screeningImage courtesy of Dr. Gavin MacBeath, Bauer Center for Genomics Research, Harvard University

  • Desirable Substrate Properties for Protein Microarray Applications Protein compatible High probe loading capacity Low inherent fluorescence and nonspecific binding background Consistent, uniform product Ease of use

  • HydroGelTM Coated Slides

  • HydroGelTM Performance Validation

    Printing compatibilityInherent fluorescent backgroundLoading capacity of substrateProtein compatibilityNonspecific backgroundMultiplexed assay performance

  • Printing CompatibilityPackard BioChip ArrayerTM (Piezo)Feature Size ~200 umPackard SpotArray 24 (Split Pin) Feature Size ~150 umHydroGelTM coated slides are compatible with both contact and non-contact printers (examples shown above). This was verified on two other types of commercially available and home-made instruments.

  • Inherent Fluorescent Background

    Blank substrates were scanned on a ScanArrayTM 5000 microarray scanner in the channels indicated (laser power:100%, PMT gain: 75%).

  • Protein Loading CapacityIgG and Streptavidin were printed on HydroGelTM coated slides and aldehyde glass slides at the indicated concentrations to compare loading capacity.

    Chart2

    9.30960561697.2019861364

    18.62119576387.0985859097

    42.416115494313.2497801135

    89.688557566618.0725385093

    136.000092104823.619003725

    HydroGel

    Aldehyde Glass

    Printing Concentration (mg/mL)

    fmoles per mm2

    Immobilization of IgG

    Sheet7

    Average of HydroGels

    SampleAvg Imm effStdevCVg immobilizedmoles immobilizedmoles/mm2fM/mm2

    TR AH IgG 0.5mg/ml48.71%3.58%7.35%0.00000000020.0000000001009.31

    TR AH IgG 1mg/ml48.71%3.50%7.18%0.00000000040.00000000020018.62

    TR AH IgG 2mg/ml55.48%2.51%4.53%0.00000000070.00000000040042.42

    TR AH IgG 4mg/ml58.66%4.35%7.42%0.00000000140.00000000080089.69

    TR AH IgG 8mg/ml44.47%5.36%12.05%0.00000000280.000000001200136.00

    TR SA 0.5mg/ml41.76%3.51%8.40%0.00000000020.00000000010021.28

    TR SA 1mg/ml44.15%3.79%8.59%0.00000000040.00000000020045.00

    TR SA 2mg/ml42.80%3.21%7.50%0.00000000070.00000000030087.26

    TR SA 4mg/ml42.87%4.67%10.90%0.00000000140.000000000600174.79

    TR SA 8mg/ml40.85%3.71%9.09%0.00000000280.000000001100333.15

    Average Telechem: AHIgGSA

    SampleAvg Imm effStdevCVg immobilizedmoles immobilizedmoles/mm2fM/mm2

    TR AH IgG 0.5mg/ml37.68%6.62%17.56%0.00000000020.0000000001007.20

    TR AH IgG 1mg/ml18.57%0.65%3.52%0.00000000040.0000000001007.10

    TR AH IgG 2mg/ml17.33%2.81%16.21%0.00000000070.00000000010013.25

    TR AH IgG 4mg/ml11.82%0.53%4.49%0.00000000140.00000000020018.07

    TR AH IgG 8mg/ml7.72%0.11%1.44%0.00000000280.00000000020023.62

    TR SA 0.5mg/ml28.72%4.00%13.92%0.00000000020.00000000010014.64

    TR SA 1mg/ml27.93%6.98%24.99%0.00000000040.00000000010028.47

    TR SA 2mg/ml23.41%3.37%14.38%0.00000000070.00000000020047.73

    TR SA 4mg/ml12.86%1.59%12.37%0.00000000140.00000000020052.42

    TR SA 8mg/ml9.44%2.27%24.08%0.00000000280.00000000030076.97

    Average of Both S&S Slides*

    SampleAvg Imm effStdevCVg immobilizedmoles immobilizedmoles/mm2fM/mm2

    TR AH IgG 0.5mg/ml78.38%7.32%9.34%0.00000000020.00000000010014.98

    TR AH IgG 1mg/ml77.65%1.21%1.55%0.00000000040.00000000030029.68

    TR AH IgG 2mg/ml75.96%2.11%2.77%0.00000000070.00000000050058.07

    TR AH IgG 4mg/ml71.20%0.64%0.90%0.00000000140.00000000100108.86

    TR AH IgG 8mg/ml72.03%6.57%9.12%0.00000000280.00000000200220.26

    TR SA 0.5mg/ml53.45%2.13%3.99%0.00000000020.00000000010027.24

    TR SA 1mg/ml56.89%1.15%2.01%0.00000000040.00000000020057.99

    TR SA 2mg/ml66.89%1.52%2.27%0.00000000070.000000000500136.38

    TR SA 4mg/ml73.82%1.68%2.28%0.00000000140.00000000100300.98

    TR SA 8mg/ml71.49%6.30%8.82%0.00000000280.00000000200582.99

    Sheet1

    Average of HydroGels 61401Average Telechem: AHIgG 53001SA 61401Average of Both S&S Slides*

    SampleAvg Imm effStdevCVg immobilizedmoles immobilizedmoles/mm2fM/mm2SampleAvg Imm effStdevCVg immobilizedmoles immobilizedmoles/mm2fM/mm2SampleAvg Imm effStdevCVg immobilizedmoles immobilizedmoles/mm2fM/mm2

    TR AH IgG 0.5mg/ml48.71%3.58%7.35%0.00000000020.0000000001009.31TR AH IgG 0.5mg/ml37.68%6.62%17.56%0.00000000020.0000000001007.20TR AH IgG 0.5mg/ml78.38%7.32%9.34%0.00000000020.00000000010014.98

    TR AH IgG 1mg/ml48.71%3.50%7.18%0.00000000040.00000000020018.62TR AH IgG 1mg/ml18.57%0.65%3.52%0.00000000040.0000000001007.10TR AH IgG 1mg/ml77.65%1.21%1.55%0.00000000040.00000000030029.68

    TR AH IgG 2mg/ml55.48%2.51%4.53%0.00000000070.00000000040042.42TR AH IgG 2mg/ml17.33%2.81%16.21%0.00000000070.00000000010013.25TR AH IgG 2mg/ml75.96%2.11%2.77%0.00000000070.00000000050058.07

    TR AH IgG 4mg/ml58.66%4.35%7.42%0.00000000140.00000000080089.69TR AH IgG 4mg/ml11.82%0.53%4.49%0.00000000140.00000000020018.07TR AH IgG 4mg/ml71.20%0.64%0.90%0.00000000140.00000000100108.86

    TR AH IgG 8mg/ml44.47%5.36%12.05%0.00000000280.000000001200136.00TR AH IgG 8mg/ml7.72%0.11%1.44%0.00000000280.00000000020023.62TR AH IgG 8mg/ml72.03%6.57%9.12%0.00000000280.00000000200220.26

    TR SA 0.5mg/ml41.76%3.51%8.40%0.00000000020.00000000010021.28TR SA 0.5mg/ml28.72%4.00%13.92%0.00000000020.00000000010014.64TR SA 0.5mg/ml53.45%2.13%3.99%0.00000000020.00000000010027.24

    TR SA 1mg/ml44.15%3.79%8.59%0.00000000040.00000000020045.00TR SA 1mg/ml27.93%6.98%24.99%0.00000000040.00000000010028.47TR SA 1mg/ml56.89%1.15%2.01%0.00000000040.00000000020057.99

    TR SA 2mg/ml42.80%3.21%7.50%0.00000000070.00000000030087.26TR SA 2mg/ml23.41%3.37%14.38%0.00000000070.00000000020047.73TR SA 2mg/ml66.89%1.52%2.27%0.00000000070.000000000500136.38

    TR SA 4mg/ml42.87%4.67%10.90%0.00000000140.000000000600174.79TR SA 4mg/ml12.86%1.59%12.37%0.00000000140.00000000020052.42TR SA 4mg/ml73.82%1.68%2.28%0.00000000140.00000000100300.98

    TR SA 8mg/ml40.85%3.71%9.09%0.00000000280.000000001100333.15TR SA 8mg/ml9.44%2.27%24.08%0.00000000280.00000000030076.97TR SA 8mg/ml71.49%6.30%8.82%0.00000000280.00000000200582.99

    TRIgGSA

    Conc (mg/ml)HydroGelTelechemS&S FASTConc (mg/ml)HydroGelTelechemS&S FAST

    0.59.317.2014.980.521.2814.6427.24

    118.627.1029.68145.0028.4757.99

    242.4213.2558.07287.2647.73136.38

    489.6918.07108.864174.7952.42300.98

    8136.0023.62220.268333.1576.97582.99

    Sheet1

    000

    000

    000

    000

    000

    HydroGel

    Telechem

    S&S FAST

    Printing Concentration (mg/ml)

    fMoles/mm2

    Sheet2

    000

    000

    000

    000

    000

    HydroGel

    Telechem

    S&S FAST

    Printing Concentration (mg/ml)

    fMoles/mm2

    redo charts for poster

    redo charts for poster

    9.30960561697.201986136414.9805292165

    18.62119576387.098585909729.6814897545

    42.416115494313.249780113558.0694768449

    89.688557566618.0725385093108.8634435824

    136.000092104823.619003725220.2603579635

    HydroGel

    Telechem

    S&S FAST

    Printing Concentration (mg/ml)

    fMoles/mm2

    IgG Immobilization

    Sheet4

    21.283652520614.636974044227.2429148501

    44.999198482228.465669920857.9923212051

    87.261465647747.7318188349136.3762587651

    174.791459764752.423923163300.9778225317

    333.148352016476.9726680627582.9896454343

    HydroGel

    Telechem

    S&S FAST

    Printing Concentration (mg/ml)

    fMoles/mm2

    Streptavidin Immobilization

    Sheet5

    HydroGelAldehyde GlassHydroGelAldehyde Glass

    0.59.317.200.521.2814.64

    118.627.10145.0028.47

    242.4213.25287.2647.73

    489.6918.074174.7952.42

    8136.0023.628333.1576.97

    Sheet5

    00

    00

    00

    00

    00

    HydroGel

    Aldehyde Glass

    Printing Concentration (mg/mL)

    fmoles per mm2

    Immobilization of IgG

    Sheet6

    00

    00

    00

    00

    00

    HydroGel

    Aldehyde Glass

    Printing Concentration (mg/mL)

    fmoles per mm2

    Immobilization of Streptavidin

    Chart3

    21.283652520614.6369740442

    44.999198482228.4656699208

    87.261465647747.7318188349

    174.791459764752.423923163

    333.148352016476.9726680627

    HydroGel

    Aldehyde Glass

    Printing Concentration (mg/mL)

    fmoles per mm2

    Immobilization of Streptavidin

    Sheet7

    Average of HydroGels

    SampleAvg Imm effStdevCVg immobilizedmoles immobilizedmoles/mm2fM/mm2

    TR AH IgG 0.5mg/ml48.71%3.58%7.35%0.00000000020.0000000001009.31

    TR AH IgG 1mg/ml48.71%3.50%7.18%0.00000000040.00000000020018.62

    TR AH IgG 2mg/ml55.48%2.51%4.53%0.00000000070.00000000040042.42

    TR AH IgG 4mg/ml58.66%4.35%7.42%0.00000000140.00000000080089.69

    TR AH IgG 8mg/ml44.47%5.36%12.05%0.00000000280.000000001200136.00

    TR SA 0.5mg/ml41.76%3.51%8.40%0.00000000020.00000000010021.28

    TR SA 1mg/ml44.15%3.79%8.59%0.00000000040.00000000020045.00

    TR SA 2mg/ml42.80%3.21%7.50%0.00000000070.00000000030087.26

    TR SA 4mg/ml42.87%4.67%10.90%0.00000000140.000000000600174.79

    TR SA 8mg/ml40.85%3.71%9.09%0.00000000280.000000001100333.15

    Average Telechem: AHIgGSA

    SampleAvg Imm effStdevCVg immobilizedmoles immobilizedmoles/mm2fM/mm2

    TR AH IgG 0.5mg/ml37.68%