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IgG Detection Using Anti- IgG Conjugated Microparticles

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

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  1. IgG Detection Using Anti-IgG Conjugated Microparticles By Sarah Leung and AnbarNajam

  2. Methods • Conjugated anti-IgG antibodies to 0.92 um microparticles • 33% microparticle surface coverage with IgG • 0.02% microparticle concentration in solution

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

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

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

  6. 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

  7. 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

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

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

  10. 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

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

  12. 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

  13. Methods • 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

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

  15. Discussion • Results oscillate and do not form a smooth curve. • Possible explanations: • Surface contamination • Droplet movement, vibration. • Position of droplet. • Size of droplet

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

  17. Methods

  18. Results Deionized Water

  19. Future Work Minimize the oscillations. Change the position of the surface. Determine an efficient way to dispense 3ul droplet on to superhydrophobic surface

  20. Acknowledgement s • 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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