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.

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Igg detection using anti igg conjugated microparticles

IgG Detection Using Anti-IgG Conjugated Microparticles

By Sarah Leung and

AnbarNajam


Methods
Methods

  • Conjugated anti-IgG antibodies to 0.92 um microparticles

  • 33% microparticle surface coverage with IgG

  • 0.02% microparticle concentration in solution


Methods1
Methods

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


Results
Results

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


Results1
Results

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


Discussion
Discussion

  • Results lacked evidence of any trends between antigen concentration and light scattering intensity

  • Possible explanations:

    • Coagulation of microparticles

    • High scattering due to 180 degree detection

    • Conjugation of antibody, rather than antigen, to microparticles


Next experiment
Next Experiment

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

Vs.

180 degrees

45 degrees


Results2
Results

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


Results3
Results

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


Discussion1
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


Methods2
Methods Conjugated

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

  • 33% microparticle surface coverage with anti-E. coli

  • 0.02% microparticle concentration in solution


Methods3
Methods Conjugated

  • Dispensed 10 ul droplet on to superhydrophobic surface.

  • Measured contact angle over 10 minutes

  • Droplets:

    • Deionized Water

    • Anti-E. coli -Conc. 5*10-7

    • Anti-E. coli conjugated to microparticles


Results4
Results Conjugated

Anti-E. coli -Conc. 5*10-7


Discussion2
Discussion Conjugated

  • Results oscillate and do not form a smooth curve.

  • Possible explanations:

    • Surface contamination

    • Droplet movement, vibration.

    • Position of droplet.

    • Size of droplet


Next experiment1
Next Experiment Conjugated

Decrease the volume of the droplet from 10 ul to 3 ul for a more spherical shape.


Methods4
Methods Conjugated


Results5
Results Conjugated

Deionized Water


Future work
Future Work Conjugated

Minimize the oscillations.

Change the position of the surface.

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


Acknowledgement s
Acknowledgement s Conjugated

  • UA/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


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