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Investigation of Constructing a Biosensor for Endotoxin Detection in Biological Products and Comparison of the Results with Conventional Endotoxin Detection Methods

Zandieh, Mohammad | 2017

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  1. Type of Document: M.Sc. Thesis
  2. Language: Farsi
  3. Document No: 49834 (06)
  4. University: Sharif University of Technology
  5. Department: Chemical and Petroleum Engineering
  6. Advisor(s): Vosoughi, Manouchehr; Hosseini, Nezameddin
  7. Abstract:
  8. Lipopolysaccharide (LPS), also known as endotoxin, is a highly toxic component exists in the outer membrane of gram negative bacteria. It releases into the environment during every phase of bacterial growth cycle, so it causes contamination of a wide range of biopharmaceutical products. Even small quantities of endotoxin injected to human body can result in fever, septic shock, and death. Therefore, it is highly important to detect and also quantify endotoxin of biopharmaceutical products in quality control laboratories. The most validated method used for endotoxin detection is Limulus Amebocyte Lysate (LAL). Although this method is sensitive, it has some unavoidable drawbacks such as highly dependency to temperature, long incubation times to access the results, hard multistep preparation procedures and being expensive. These problems forced research groups all around the world into working on novel methods of endotoxin detection based on biosensors. In recent years, LSPR biosensors are gaining importance due to their interesting detection characteristics such as simplicity and sensitivity. This thesis describes the construction of a silver-based LSPR biosensor for lipopolysaccharide (LPS) detection. A polycationic antibiotic, Polymyxin B, has been used as a biological element to interact with endotoxin. In this work, the silver nanostructures were considerably stabilized by a self-assembled monolayer (SAM) of MPA, and the limit of detection of biosensor was measured to be 340 pg/ml. We also investigated stability, reproducibility and selectivity of the biosensor in various experiments
  9. Keywords:
  10. Quality Control ; Biosensor ; Endotoxin Bacterium ; Localized Surface Plasmons ; Biological Products ; Pharmaceutical Biotechnology

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