Transcript
Page 1: IgG  Detection Using Anti- IgG  Conjugated  Microparticles

IgG Detection Using IgG Detection Using Anti-IgG Conjugated Anti-IgG Conjugated

MicroparticlesMicroparticles

By Sarah Leung and By Sarah Leung and

Anbar NajamAnbar Najam

Page 2: IgG  Detection Using Anti- IgG  Conjugated  Microparticles

MethodsMethods

Conjugated anti-IgG antibodies to 0.92 Conjugated anti-IgG antibodies to 0.92 um microparticles um microparticles

33% microparticle surface coverage with 33% microparticle surface coverage with IgGIgG

0.02% microparticle concentration in 0.02% microparticle concentration in solutionsolution

Page 3: IgG  Detection Using Anti- IgG  Conjugated  Microparticles

MethodsMethods

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

Page 4: IgG  Detection Using Anti- IgG  Conjugated  Microparticles

ResultsResults 5*105*10-11 -11 g/mL to 5*10g/mL to 5*10-6 -6 g/mL IgG serial dilutionsg/mL IgG serial dilutions

12600

12700

12800

12900

13000

13100

13200

13300

13400

13500

13600

13700

1E-12 1E-10 1E-08 0.000001 0.0001 0.01 1

Concentration (g/mL)

Inte

nsi

ty

Page 5: IgG  Detection Using Anti- IgG  Conjugated  Microparticles

ResultsResults5*105*10-15 -15 g/mL to 5*10g/mL to 5*10-6 -6 g/mL IgG serial dilutionsg/mL IgG serial dilutions

13400.00

13600.00

13800.00

14000.00

14200.00

14400.00

14600.00

14800.00

15000.00

15200.00

1E-17 1E-14 1E-11 0.00000001 0.00001 0.01 10

Concentration (g/mL)

Inte

ns

ity

Page 6: IgG  Detection Using Anti- IgG  Conjugated  Microparticles

DiscussionDiscussion

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

Possible explanations:Possible explanations: Coagulation of microparticlesCoagulation of microparticles High scattering due to 180 degree detectionHigh scattering due to 180 degree detection Conjugation of antibody, rather than antigen, Conjugation of antibody, rather than antigen,

to microparticlesto microparticles

Page 7: IgG  Detection Using Anti- IgG  Conjugated  Microparticles

Next ExperimentNext Experiment

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

180 degrees 45 degreesVs.

Page 8: IgG  Detection Using Anti- IgG  Conjugated  Microparticles

ResultsResults

5*105*10-14 -14 g/mL to 5*10g/mL to 5*10-6 -6 g/mL IgG serial dilutionsg/mL IgG serial dilutions

Page 9: IgG  Detection Using Anti- IgG  Conjugated  Microparticles

ResultsResults

5*105*10-14 -14 g/mL to 5*10g/mL to 5*10-6 -6 g/mL IgG serial dilutionsg/mL IgG serial dilutions

Page 10: IgG  Detection Using Anti- IgG  Conjugated  Microparticles

DiscussionDiscussion

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

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

Page 11: IgG  Detection Using Anti- IgG  Conjugated  Microparticles

Contact Angle Measurements of Contact Angle Measurements of Anti-E. Coli and Anti-E. Coli Anti-E. Coli and Anti-E. Coli

Conjugated Microparticles on Conjugated Microparticles on Superhydrophobic SurfaceSuperhydrophobic Surface

Page 12: IgG  Detection Using Anti- IgG  Conjugated  Microparticles

MethodsMethods

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

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

0.02% microparticle concentration in solution0.02% microparticle concentration in solution

Page 13: IgG  Detection Using Anti- IgG  Conjugated  Microparticles

MethodsMethods

Dispensed 10 ul droplet on to superhydrophobic Dispensed 10 ul droplet on to superhydrophobic surface.surface.

Measured contact angle over 10 minutesMeasured contact angle over 10 minutes

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

Page 14: IgG  Detection Using Anti- IgG  Conjugated  Microparticles

ResultsResults

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

Page 15: IgG  Detection Using Anti- IgG  Conjugated  Microparticles

DiscussionDiscussion

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

Possible explanations:Possible explanations: Surface contaminationSurface contamination Droplet movement, vibration.Droplet movement, vibration. Position of droplet.Position of droplet. Size of dropletSize of droplet

Page 16: IgG  Detection Using Anti- IgG  Conjugated  Microparticles

Next ExperimentNext Experiment

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

Page 17: IgG  Detection Using Anti- IgG  Conjugated  Microparticles

Methods

Page 18: IgG  Detection Using Anti- IgG  Conjugated  Microparticles

ResultsResults

Deionized WaterDeionized Water

Page 19: IgG  Detection Using Anti- IgG  Conjugated  Microparticles

Future WorkFuture Work

Minimize the oscillations.Minimize the oscillations.

Change the position of the surface.Change the position of the surface.

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

Page 20: IgG  Detection Using Anti- IgG  Conjugated  Microparticles

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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