High-Throughput Agglutination Assays: Two-Phase Microfluidic Systems, Slides of Engineering

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TWO-PHASE MICROFLUIDIC

SYSTEMS FOR HIGH THROUGHPUT

QUANTIFICATION OF

AGGLUTINATION ASSAYS

ENGR 509 Microfluidics

BY: ABDUL BASIT ZIA

2 DIAGNOSTIC TECHNIQUES- IV Agglutination

Agglutination reactions produce visible aggregates of antibody –

antigen complexes when antibodies or antigens are conjugated

to a carrier.

Classification of Immunoassays:

• (^) Qualitative vs Quantitative
• (^) Target Analyte Type
• (^) Labeled vs Non labeled
• (^) Homogenous vs Heterogeneous

4 ABO, D BLOOD TYPING AND SUBTYPING USING PLUG-BASED MICROFLUIDICS [2]

• (^) Droplets size of 30-40nL
• (^) Combine streams of antibody, buffer, RBCs
• (^) Image processing for quantification of agglutination
  • (^) Images taken and analyzed manually [2] T. R. Kline et al , "ABO, D Blood Typing and Subtyping Using Plug-Based Microfluidics," Anal. Chem., vol. 80, (16), pp. 6190-6197, 08/01, 2008.

5 MICROINJECTION MOLDED DISPOSABLE MICROFLUIDIC LAB-ON-A-CHIP FOR EFFICIENT DETECTION OF AGGLUTINATION [3]

• (^) Complete lab-on-chip was realized by microinjection molding cyclic olefin copolymer (COC)
• (^) Integrated 1) inlet microwells
  2. Improved serpentine laminating micromixer (ISLM) 3)Reaction well
• (^) Microchannel width 250 μm
• (^) Passive valve – channel width 100μm
• COC substrate is replicated within 1 minute
• (^) Simulated blood (WARD’s ™) with simulated sera (Anti –A/B) [3] - S. H. Choi, D. S. Kim and T. H. Kwon, "Microinjection molded disposable microfluidic lab-on-a-chip for efficient detection of agglutination," Microsystem Technologies, vol. 15, (2), pp. 309-316, 02/01, 2009.

7 QUANTIFICATION OF A LATEX AGGLUTINATION ASSAY FOR BACTERIAL PATHOGEN DETECTION IN A LOW-COST CAPILLARY-DRIVEN FLUIDIC PLATFORM [5]

• (^) Latex agglutination – for Escherichia coli (E. Coli.) bacteria
• (^) Direct Testing – latex particles covered by antibodies
• (^) Fluid driving force: Capillary forces
• (^) Fluid channel depth 140 μm
• (^) lower detection limit on the order of 10^6 cells/mL
• (^) Conventional lower detection limit of 10^7 cells/mL [5] - O. S. Pak, K. Pietrzyk, A. Ly, A. Maldonado-Liu, S. Fukuoka and U. Kim, "Quantification of a latex agglutination assay for bacterial pathogen detection in a low-cost capillary- driven fluidic platform," 2016 IEEE Global Humanitarian Technology Conference (GHTC) , Seattle, WA, 2016, pp. 765-769. doi: 10.1109/GHTC.2016.

8 CAPACITIVE IMMUNOSENSOR FOR C-REACTIVE PROTEIN QUANTIFICATION [6]

• (^) Non labeled immunoassays
• (^) Low detection limit 0.5 mg/L
• (^) Gold interdigitated electrodes (IDEs)
• (^) LCR meter used at 1MHz at 1V [6] - C. Sapsanis, S. Sivashankar, H. Omran, U. Buttner and K. N. Salama, "Capacitive immunosensor for C-reactive protein quantification," 2015 IEEE 58th International Midwest Symposium on Circuits and Systems (MWSCAS) , Fort Collins, CO, 2015, pp. 1-4. doi: 10.1109/MWSCAS.2015.

Detail

Questions

11 Back

Prozone phenomenon is a false negative

response resulting from high antibody titer which

interferes with formation of antigen- antibody

lattice, necessary to visualize

13