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Pharmaceutical technology assignment.

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Proteomics, Bio-molecular Identification and Gene therapy

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Pharmaceutical technology assignment.

  1. 1. PROTEOMICS, BIOMOLECULAR IDENTIFICATION AND GENE THERAPY Prepared by: Waqar , Sarmad, Amir, Rizwan & Junaid PHARMACISTS OF SHAH ABDUL LATIF UNIVERSITY KHAIRPUR SINDH,PAKISTAN
  2. 2. INTRODUCTION TO PROTEOMICS Proteomics is simply defined as the Study of Proteome. The term proteome was created by Wasinger et al. (1995) and Wilkins et al. (1996) and defined as the cell- and tissue-specific patterns of proteins expressed by the genome of an organism. Proteomics is also defined as the study of total protein content of a cell or that of an organism.
  3. 3. SCOPE OF PROTEOMICS  Proteomics helps in understanding the alteration in protein expression during different stages of life cycle or under stress condition.  Proteomics is also important with regards to drug development against several diseases and understanding various biological processes, as proteins are the most favourable targets for various drugs.
  4. 4. SCOPE OF PROTEOMICS  Proteomics helps in understanding the structure and function of different proteins as well as protein-protein interactions of an organism and a minor defect in either protein structure, its function or alternation in expression pattern can be easily detected using proteomics studies.  Protein to gene can be predicted by Proteomics.
  5. 5. CLASSIFICATION OF PROTEOMICS 1. Functional Proteomics: Identifications of Protein-Protein, Protein-DNA and Protein-RNA interactions affecting function. 2. Structural Proteomics: Identification of all interactions by metal ions, toxins, drugs etc. affecting Protein structure. 3. Differential Proteomics: Determination of differences in Protein expression.
  6. 6. BIO-MOLECULAR IDENTIFICATION Initial step in all proteomic studies is the separation of proteins mixture by Two Dimensional Gel Electrophoresis technique, based on their individual charges in 1D. The gel is then turned 90 degrees to separate proteins based on the difference in their size. This separation occurs in 2nd dimension hence the name 2D. The spots obtained in 2D electrophoresis are excised and further subjected to mass spectrometric analysis of each protein present in the mixture.
  7. 7. STEPS IN PROTEOMIC ANALYSIS  Purification of proteins: It involves extraction of protein sample followed by purification using density gradient centrifugation, chromatographic techniques (affinity etc.)  Separation of proteins: 2D gel electrophoresis is applied for separation of proteins on the basis of their isoelectric points in one dimension and molecular weight on the other. Spots are detected using fluorescent dyes or radioactive probes.
  8. 8. STEPS IN PROTEOMIC ANALYSIS  Identification of proteins: The separated protein spots on gel are excised and digested in gel by a protease (e.g. trypsin). The eluted peptides are identified using mass spectrometry.  Validation of Proteins: Determined amino acid sequence is finally compared with available database to validate the proteins.
  9. 9. OVERVIEW OF STEPS
  10. 10. TECHNIQUES INVOLVED IN PROTEIN IDENTIFICATION
  11. 11. GENE THERAPY Pathological changes within an organism arise from protein alterations. Proteomics provides a powerful set of tools for the large-scale study of protein expression. The identification of such changes by proteomics is essential to define drug targets, therapeutic proteins, and disease biomarkers. Gene therapy, primarily based on the delivery of a “therapeutic” gene to the diseased organ, is an approach whose effect is finally due to the function of the therapeutic protein encoded by the
  12. 12. GENE THERAPY “therapeutic” gene, proteomics provides the basis required for the design and application of gene therapies. Because in certain diseases the absence of certain cellular or biological factors, for example Enzymes occur and it is may be due to a gene being defective. Cancer, infectious diseases, cardiovascular diseases, neurological disorders and some inherited conditions are among the areas into which the Gene therapy research is being carried out.
  13. 13. APPLICATIONS OF PROTEOMICS 1. For discovery of protein biomarkers. 2. In the Study of Tumor Metastasis. 3. In the Field of Neuro-trauma. 4. In disease diagnosis. 5. In Neurology. 6. In Treatment of Autoimmune disease. 7. In Nutrition Research.

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