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1
Dual Detection High-Speed Capillary
Electrophoresis (HSCE)
By: Lamees Zaro
Dr. Rasmi Abu Helu
Al-Quds University
Faculty of Graduate Studies
Department of Medical Laboratory Sciences
12/21/2024
2
What is Capillary Electrophoresis?
 A technique that separates molecules, such
as proteins, based on their size and charge
when subjected to an electric field.
 It provides higher efficiency and throughput.
12/21/2024
3
What is Serum Protein Electrophoresis (SPE)?
 Serum proteins are separated based on their
charge-to-mass ratio using an electric field.
 Major serum proteins albumin, 留1, 留2, 硫 , 粒 are
separated.
Abnormal distributions are often indicative of disease.
 The Refractive Index (RI) method is used to detect
the proteins' migration through the capillary.
12/21/2024
4
What is an Immunoassay?
 Immunoassays are based on the specific binding between antigens and
antibodies.
 This interaction forms an antigen-antibody complex, which can be detected
by fluorescence.
 Immunoassay Detection in Capillary Electrophoresis:
 The antigen-antibody complex is separated from the free antigen based on
size and charge, and fluorescence detection is used to quantify the complex.
12/21/2024
5
What is Dual Detection HSCE?
 HSCE combines two detection methods:
1. Refractive Index (RI) detection for serum protein electrophoresis.
2. Fluorescence detection for immunoassay measurements.
This enables the simultaneous measurement of serum proteins and antigen-
antibody interactions in one analysis.
Speed and Efficiency.
Higher Sensitivity and Specificity.
12/21/2024
6
Schematic of the dual detection HSCE platform
Opallage, P. M., De Silva, M., &
Dunn, R. C. (2022)
12/21/2024
7
Serum Protein Electropherograms (SPE) Measured In
Various Buffers Using HSCE Combined With BSI Detection.
Opallage, P. M., De Silva, M., &
Dunn, R. C. (2022)
12/21/2024
8
Evaluates Buffer Performance For Antigen-antibody Interaction Detection
Opallage, P. M., De Silva, M., &
Dunn, R. C. (2022)
12/21/2024
9
Simultaneous SPE And Fluorescence Detection For Three Different Antibody Levels.
Opallage, P. M., De Silva, M., &
Dunn, R. C. (2022)
12/21/2024
10
Opallage, P. M., De Silva, M., &
Dunn, R. C. (2022)
Fluorescence Electropherograms Across Antibody Concentrations
12/21/2024
11
Calibration Curve for Immunoassay Quantification
Opallage, P. M., De Silva, M., &
Dunn, R. C. (2022)
12/21/2024
12
Benefits
High sensitivity and resolution.
Rapid and automated analysis.
Capability to detect both qualitative and quantitative abnormalities in serum and
urine.
12/21/2024
13
Applications
 Clinical diagnostics.
1. Protein disorders.
2. Immune system disorders.
3. Infectious diseases.
 Research applications.
 Pharmaceutical applications (therapeutic antibodies, vaccine development, drug
monitoring).
12/21/2024
14
12/21/2024

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Electrophoretic Technique Electro .pptx

  • 1. 1 Dual Detection High-Speed Capillary Electrophoresis (HSCE) By: Lamees Zaro Dr. Rasmi Abu Helu Al-Quds University Faculty of Graduate Studies Department of Medical Laboratory Sciences 12/21/2024
  • 2. 2 What is Capillary Electrophoresis? A technique that separates molecules, such as proteins, based on their size and charge when subjected to an electric field. It provides higher efficiency and throughput. 12/21/2024
  • 3. 3 What is Serum Protein Electrophoresis (SPE)? Serum proteins are separated based on their charge-to-mass ratio using an electric field. Major serum proteins albumin, 留1, 留2, 硫 , 粒 are separated. Abnormal distributions are often indicative of disease. The Refractive Index (RI) method is used to detect the proteins' migration through the capillary. 12/21/2024
  • 4. 4 What is an Immunoassay? Immunoassays are based on the specific binding between antigens and antibodies. This interaction forms an antigen-antibody complex, which can be detected by fluorescence. Immunoassay Detection in Capillary Electrophoresis: The antigen-antibody complex is separated from the free antigen based on size and charge, and fluorescence detection is used to quantify the complex. 12/21/2024
  • 5. 5 What is Dual Detection HSCE? HSCE combines two detection methods: 1. Refractive Index (RI) detection for serum protein electrophoresis. 2. Fluorescence detection for immunoassay measurements. This enables the simultaneous measurement of serum proteins and antigen- antibody interactions in one analysis. Speed and Efficiency. Higher Sensitivity and Specificity. 12/21/2024
  • 6. 6 Schematic of the dual detection HSCE platform Opallage, P. M., De Silva, M., & Dunn, R. C. (2022) 12/21/2024
  • 7. 7 Serum Protein Electropherograms (SPE) Measured In Various Buffers Using HSCE Combined With BSI Detection. Opallage, P. M., De Silva, M., & Dunn, R. C. (2022) 12/21/2024
  • 8. 8 Evaluates Buffer Performance For Antigen-antibody Interaction Detection Opallage, P. M., De Silva, M., & Dunn, R. C. (2022) 12/21/2024
  • 9. 9 Simultaneous SPE And Fluorescence Detection For Three Different Antibody Levels. Opallage, P. M., De Silva, M., & Dunn, R. C. (2022) 12/21/2024
  • 10. 10 Opallage, P. M., De Silva, M., & Dunn, R. C. (2022) Fluorescence Electropherograms Across Antibody Concentrations 12/21/2024
  • 11. 11 Calibration Curve for Immunoassay Quantification Opallage, P. M., De Silva, M., & Dunn, R. C. (2022) 12/21/2024
  • 12. 12 Benefits High sensitivity and resolution. Rapid and automated analysis. Capability to detect both qualitative and quantitative abnormalities in serum and urine. 12/21/2024
  • 13. 13 Applications Clinical diagnostics. 1. Protein disorders. 2. Immune system disorders. 3. Infectious diseases. Research applications. Pharmaceutical applications (therapeutic antibodies, vaccine development, drug monitoring). 12/21/2024