IgG Detection Using Anti- IgG Conjugated Microparticles

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IgG Detection Using Anti- IgG Conjugated Microparticles. By Sarah Leung and Anbar Najam. Methods. Conjugated anti-IgG antibodies to 0.92 um microparticles 33% microparticle surface coverage with IgG 0.02% microparticle concentration in solution. Methods. - PowerPoint PPT Presentation

Text of IgG Detection Using Anti- IgG Conjugated Microparticles

  • IgG Detection Using Anti-IgG Conjugated MicroparticlesBy Sarah Leung and Anbar Najam

  • Methods Conjugated anti-IgG antibodies to 0.92 um microparticles

    33% microparticle surface coverage with IgG

    0.02% microparticle concentration in solution

  • Methods Positioned a 180 degree optical reflection probe directly above samples for detection on a two-well slide

  • Results 5*10-11 g/mL to 5*10-6 g/mL IgG serial dilutions

    Chart1

    12781

    13317.01

    13434.23

    13220.23

    13609.68

    12725.56

    13262.83

    Concentration (g/mL)

    Intensity

    Sheet1

    Concentration (g/mL)IntensityAverage IntensityStandard DeviationT-TestHalf Standard Deviation

    014343.2313823.9014397.2914188.14316.59707158.2985345005

    013588.0115013.8115013.8114538.54823.186010.60546411.5930069053

    014091.4413949.9114787.0014276.12448.061400.68306224.0307012629

    013554.3913991.2813768.1613771.28218.461670.29406109.2308372592

    014015.1813696.3713864.8613858.80159.491270.1062679.7456369234

    014614.2314335.9714818.3214589.51242.123550.02932121.0617745546

    0.000000000514202.5913778.8513541.3913840.94334.944840.30762167.4724205155

    0.00000000514060.2713669.9314155.5613961.92257.320160.02712128.6600813189

    0.0000000513859.6114368.3613688.4713972.15353.639620.63187176.8198087272

    0.000000514525.5713819.2113730.6614025.15435.635070.59152217.8175326008

    0.00000513982.3814030.3713884.6013965.7874.288700.4165337.1443475961

    Sheet1

    158.2985345005158.2985345005

    411.5930069053411.5930069053

    224.0307012629224.0307012629

    109.2308372592109.2308372592

    79.745636923479.7456369234

    121.0617745546121.0617745546

    167.4724205155167.4724205155

    128.6600813189128.6600813189

    176.8198087272176.8198087272

    217.8175326008217.8175326008

    37.144347596137.1443475961

    Concentration (g/mL)

    Intensity

    Sheet2

    Concentration (g/mL)

    012781

    013317.01

    0.000000000513434.23

    0.00000000513220.23

    0.0000000513609.68

    0.000000512725.56

    0.00000513262.83

    Sheet2

    Concentration (g/mL)

    Intensity

    Sheet3

  • Results5*10-15 g/mL to 5*10-6 g/mL IgG serial dilutions

    Chart2

    14188.14158.2985345005158.2985345005

    14538.5433333333411.5930069053411.5930069053

    14276.1166666667224.0307012629224.0307012629

    13771.2766666667109.2308372592109.2308372592

    13858.803333333379.745636923479.7456369234

    14589.5066666667121.0617745546121.0617745546

    13840.9433333333167.4724205155167.4724205155

    13961.92128.6600813189128.6600813189

    13972.1466666667176.8198087272176.8198087272

    14025.1466666667217.8175326008217.8175326008

    13965.783333333337.144347596137.1443475961

    Concentration (g/mL)

    Intensity

    Sheet1

    Concentration (g/mL)IntensityAverage IntensityStandard DeviationT-TestHalf Standard Deviation

    014343.2313823.9014397.2914188.14316.59707158.2985345005

    013588.0115013.8115013.8114538.54823.186010.60546411.5930069053

    014091.4413949.9114787.0014276.12448.061400.68306224.0307012629

    013554.3913991.2813768.1613771.28218.461670.29406109.2308372592

    014015.1813696.3713864.8613858.80159.491270.1062679.7456369234

    014614.2314335.9714818.3214589.51242.123550.02932121.0617745546

    0.000000000514202.5913778.8513541.3913840.94334.944840.30762167.4724205155

    0.00000000514060.2713669.9314155.5613961.92257.320160.02712128.6600813189

    0.0000000513859.6114368.3613688.4713972.15353.639620.63187176.8198087272

    0.000000514525.5713819.2113730.6614025.15435.635070.59152217.8175326008

    0.00000513982.3814030.3713884.6013965.7874.288700.4165337.1443475961

    Sheet1

    158.2985345005158.2985345005

    411.5930069053411.5930069053

    224.0307012629224.0307012629

    109.2308372592109.2308372592

    79.745636923479.7456369234

    121.0617745546121.0617745546

    167.4724205155167.4724205155

    128.6600813189128.6600813189

    176.8198087272176.8198087272

    217.8175326008217.8175326008

    37.144347596137.1443475961

    Concentration (g/mL)

    Intensity

    Sheet2

    Concentration (g/mL)

    012781

    013317.01

    0.000000000513434.23

    0.00000000513220.23

    0.0000000513609.68

    0.000000512725.56

    0.00000513262.83

    Sheet2

    Concentration (g/mL)

    Intensity

    Sheet3

  • Discussion Results lacked evidence of any trends between antigen concentration and light scattering intensity Possible explanations:Coagulation of microparticlesHigh scattering due to 180 degree detectionConjugation of antibody, rather than antigen, to microparticles

  • Next Experiment Added TWEEN 80 to existing 0.92 um microparticle solution and evaluate 180 degree vs. 45 degree light scattering detection

    180 degrees45 degreesVs.

  • Results5*10-14 g/mL to 5*10-6 g/mL IgG serial dilutions

  • Results5*10-14 g/mL to 5*10-6 g/mL IgG serial dilutions

  • Discussion 180 degree results lacked evidence of any trends between antigen concentration and light scattering intensity

    With 45 degree probe, we found a detection limit of 10-6 g/ml

  • Contact Angle Measurements of Anti-E. Coli and Anti-E. Coli Conjugated Microparticles on Superhydrophobic Surface

  • Methods Conjugated anti-E. coli antibodies to 0.92 um microparticles

    33% microparticle surface coverage with anti-E. coli

    0.02% microparticle concentration in solution

  • Methods Dispensed 10 ul droplet on to superhydrophobic surface.

    Measured contact angle over 10 minutes

    Droplets:Deionized WaterAnti-E. coli -Conc. 5*10-7 Anti-E. coli conjugated to microparticles

  • ResultsAnti-E. coli -Conc. 5*10-7

  • Discussion Results oscillate and do not form a smooth curve.

    Possible explanations:Surface contaminationDroplet movement, vibration.Position of droplet.Size of droplet

  • Next ExperimentDecrease the volume of the droplet from 10 ul to 3 ul for a more spherical shape.

  • Methods

  • ResultsDeionized Water

  • Future WorkMinimize the oscillations.

    Change the position of the surface.

    Determine an efficient way to dispense 3ul droplet on to superhydrophobic surface

  • AcknowledgementsUA/NASA Space Grant

    Dr. Jeong-Yeol Yoon

    Lab group: Tremaine B. Powell, Jin-Hee Han, Brian C. Heinze, Phat L. Tran, Jennine N. Chesler, Dr. Keesung Kim, and Dr. Song