Electrophoretic Technique Electro .pptx

Electrophoretic Technique

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 1

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 2

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, α 1, α 2, β , γ 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 3

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 4

5. What is Dual Detection HSCE? HSCE combines two detection methods: Refractive Index (RI) detection for serum protein electrophoresis. 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 5

6. Schematic of the dual detection HSCE platform Opallage, P. M., De Silva, M., & Dunn, R. C. (2022) 12/21/2024 6

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 7

8. Evaluates Buffer Performance For Antigen-antibody Interaction Detection Opallage, P. M., De Silva, M., & Dunn, R. C. (2022) 12/21/2024 8

9. Simultaneous SPE And Fluorescence Detection For Three Different Antibody Levels. Opallage, P. M., De Silva, M., & Dunn, R. C. (2022) 12/21/2024 9

10. Opallage, P. M., De Silva, M., & Dunn, R. C. (2022) Fluorescence Electropherograms Across Antibody Concentrations 12/21/2024 10

11. Calibration Curve for Immunoassay Quantification Opallage, P. M., De Silva, M., & Dunn, R. C. (2022) 12/21/2024 11

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 12

13. Applications Clinical diagnostics. Protein disorders. Immune system disorders. Infectious diseases. Research applications. Pharmaceutical applications (therapeutic antibodies, vaccine development, drug monitoring). 12/21/2024 13

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