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VEGF Isolation from Platelet Lysate

Sanaei, Reza | 2017

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  1. Type of Document: M.Sc. Thesis
  2. Language: Farsi
  3. Document No: 49240 (06)
  4. University: Sharif University of Technology
  5. Department: Chemical and Petroleum Engineering
  6. Advisor(s): Abdekhodaie, Mohammad Jafar
  7. Abstract:
  8. Platelet-rich plasma as a juvenile to the narrow repertoire of orthopedic medicine, has risen hope to attain a generic autologous regenerative formula for different conditions. Vast plethora of GFs, cytokines and chemokines has endowed PRP with excellent regenerative properties, however not all of them would favor different conditions. Vascular endothelial growth factor has been demonstrated to be involved in the progress of Osteoarthritis by amplifying the catabolic processes which deteriorate the extracellular matrix of cartilage. Though cumbersome, the result of eliminating VEGF from platelet lysate would be promising. VEGF isolation from platelet lysate requires selectivity. By employing Bevacizumab, an anti-cancer monoclonal antibody drug, VEGF was captured from platelet lysate by immunoprecipitation. This requires immobilization of antibodies on the surface of a support, and optimization of immobilization and separation conditions. Silicagel particles with 45-75 micrometer size and 300 A¬¬o¬¬ pore size, were utilized as support. Covalent immobilization of Abs, requires functional groups on the silica particles, which was prepared by aminosilanization process. Zero-length crosslinking by EDC/NHS amidation was exploited to attach mAbs to the support. Accordingly, suitable conditions of immobilization such as pH and the concentration of crosslinking reagents was inquired. Thenceforth separation was conducted and the analysis of immunosorbent’s selectivity and recovery capacity from real samples were performed using ELISA. Recovery percentage of VEGF from diluted PL and desorption percentage stood at 82% and 75%, respectively. These values are significant in respect to relevant studies, therefore the procedure used in this project can be utilized in isolation of similar analytes from real biological samples
  9. Keywords:
  10. Vascular Endothelial Growth Factor (VEGF) ; Osteoarthritis ; Platelet Lysate ; Mono Clonl Antibody Immobilization ; Immunoprecipitaion

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