Construction and design of a novel drug delivery system


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Over the years upon the evolution in the field of nanotechnology various materials are introduced and its applications are being explored in reality proving its usefulness. Applications of the same in drug/gene delivery systems is being carried out since past decade and unbelievable results are being achieved. However, problem lying in these techniques being either in cellular uptake of the material or the degradability of the material being used. My work shall concentrate upon introducing a biocompatible and inert systems with molecules fabricated on the surface triggering cellular intake. Genetic system implemented in the system shall rely upon the mechanism of Viral genes assisted integration of the "GOI (Gene of Interest)" into the target specific location in the nucleus. And the GOI is also tagged with the Green Fluorescent Protein expressing Gene so as to confirm the presence of integration of GOI into the genome. This method shall provide a fast , reliable and non-invasive method of tracking down the delivery system from the point of injection to until it delivers the cargo.

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Construction and design of a novel drug delivery system

  1. 1. BALAGANESH KURUBA second year , masters in biology department of biological, physical and chemical sciences Illinois institute of technology Bionanotechnology Mentor:Prof. David Gidalevitz, Departmentof Physics, Illinois Instituteof Technology
  2. 2. CONTENTS Introduction Need for Drug/Gene Delivery systems Problems faced by existing Drug delivery systems Gold Nanospheres Genetic construct Fabrication Detection Future prospects Conclusion
  3. 3. INTRODUCTION • Several approaches are designed to produce materials at “Nano” scale to increase their prowess and efficiency. • Used in several aspects in the fields on research and manufacture as in production of IC’s, design of motors or transistors performing intense and complicated functions. • Building up nanobots – surveillance, bioremediation and bio/nuclear hazard detection. • Nanomaterials – Imposing immense strength and flexibility into structures and reducing the size concerns. • Storage – Infinite opportunities in storing vast amount of data in terms of TB/µm.
  4. 4. • Drug Delivery systems: Liposomes, Hydrogels and others. • Diagnostic applications: Use of Quantum dots, CLIOs and gold nanoparticles in detection of potential threats. • Nano scaffolds: Supports by nanowires or Carbon nanotubes for generating tissues via the implementation of stem cells. Eg: Bone calcification, muscle tissue generation. • High throughput Analysis: Multiplex screening of drugs/ other target molecules in ligand selection . • Structural and behavioral studies: Nanotechnology enables the study of surface of membranes and other membraned structures when in the process of interaction so as to determine the rate of dissociation of the ligand from the receptors. Similarly, it also enables study of proteomes of different organisms. • APPLICATIONS IN THE FIELD OF LIFE SCIENCES.
  5. 5. PROBLEMS FACED BY EXISTING DRUG /GENE DELIVERYSYSTEMS • Solubility issues. • Biocompatibility and Biodegradability of the delivery system being used. • Immunoglobulin conjugated delivery systems faced an issue of short half life of Mab’s being used which determine the specificity of the system. • Viral vector systems though efficient the chances of resurrection of the vector systems have its own chances resulting in problematic conditions. • Cytotoxicity of the nanoparticles being used is another concern.
  6. 6. • Gold nanospheres are cluster of gold nanoparticles which by themselves are powerful nanomaterials. • The properties of Gold nanoparticles differ from gold nanospheres in terms of Surface Enhanced Raman scattering. • Surface Enhanced Raman Scattering SERS is a productive effect of Surface Plasmon resonance caused due to the oscillations of adjacent atoms induced by incident beam of predetermined wavelength resulting in an electromagnetic field producing SERS. • The scattering patterns vary from a solid Gold nanoparticle to a hollow gold nanosphere, which forms the basis for our project. GOLD NANOSPHERES Fig: Illustration of Generating SERS response. (Source [1])
  7. 7. • Gold nanospheres are produced a result of deposition of gold nanoparticles on a solid/solid like template (Gelatin or Latex beads) which is dissolved so as to retain the deposition of the particles. • Size and thickness of the sphere shall depend upon the diameter of the beads used and concentration of the Gold salt being used. • Process of Synthesizing: Sacrificial Galvanic replacement method. Au3+ + Ag+ Au particle + 3Ag+ ions Fig: A Computer generated model of golden nanorods assembling into a hollow sphere. (Source [2])
  8. 8. According to Sacrificial Galvanic replacement method, • Exploits the phenomenon of Redox potential between Gold (salt solution) and silver (metallic). • Use of capping agents such as citric acid determines the number of nucleation sites being formed and in turn nucleation sites determines the size of the particle being produced. • A stabilizing agent also has to be added during the process to stabilize the nucleated sites from cross reacting. • As an alternate considering the economics of the process, use of Cobalt sacrificial template is used here instead of silver. • Capping agent : Citrate • Reducing agent : Sodium borohydride • Starting material: Chloroauric acid.
  9. 9. GENETICCONSTRUCT • The Drug/Gene to be delivered can be encapsulated into the spheres by incorporating the drug in the medium prior to fabrication of the surface. • The Important step in Gene delivery being designing the construct so as to contain effective elements for insertion of Gene of Interest. • Our Construct shall also include the gene for Green fluoroscence protein ( for in vitro studies) and gene for trp b – Beta subunit of trp synthase. • Purpose of incorporating marker genes is to validate the process of integration of Gene of Interest into the human genome.
  10. 10. VIRAL ELEMENTS TO BE INCORPORATED INTO THE GENETIC CONSTRUCT • Promoter and a Poly Adenylation signal. • Viral Packaging signal ( Ѱ or €) • Reverse Transcription signals- • Polypurine tract (PPT). • t – RNA binding site (PBS) The Insert is designed depending upon the kind of viral system in use. As some use splicing patterns to express a gene in that case the insert should be contained with introns, else the introns should be got rid of. Fig: Schematic representation of the construct with necessary viral elements as indicated. (Source [3])
  11. 11. FABRICATION • The Gold Nanospheres are subjected for acetate wash prior to the process of fabrication so that the surface can readily accept the proteins/ ligand molecules being fabricated with. • Two are the kinds of molecules to be fabricated on the surface. • Ligand/Mab/Protein molecule specific to the marker on the infected cell . • The Molecule “Epsin”: mediating Clathrin - mediated endocytosis. Upon Gaining entry, the nanosphere shall open up based upon the osmotic potential of increased potassium ion concentration in the cells. Or it is feasible to open up the sphere by mild application of heat. Fig: Illustration of Clathrin mediated Endocytosis. Source [4])
  12. 12. SERS DETECTION • SERS can be detected by conventional methodologies involve use of high end lasers and analytical systems. • A Novel technique has been designed where in an optical probe emits a beam of predetermined wavelength and followed by which the receptor collects the emitted signals from SERS active molecules. • 1- Optic fiber probe; 2 – Core; 3- Shell; 4- First tapered region; 5-Second tapered region; 6-third tapered region. Fig: Schematic representation of SERS detecting probe. (Source [5])
  13. 13. REFERENCES: LITERATURE • 1. Aggelliki K. "Quantum Dots Advantages and Disadvantages." •2. Sperling R A and et al., "Biological Applications of Gold Nanoparticles". Chem Soc Rev 2008. •3. Hainfield JF and et al., "Gold nanoparticles: a new X ray contrast agent". British Journal of Radiology 2006. •4. Pissuwan D and et al., "Therapeutic possibilities of plasmonically heated Gold Nanoparticles." Trends Biotechnol 2006. •5. Cullen T Vogelson. " Advances in Drug delivery systems." ACS publications April 2001. •6. Ghosh P and et al., " Gold Nanoparticles in Delivery applications" . Adv Drug Delivery April 2008. •7. Chen C Kwak and et al., "Packaging of viral capsid using gold nanoparticles". Journal of Nanoscience and Nanotechnology 2008. •8. Quinten M. Appl. Physics B.2001, 73, 317. •9. Chen J Y and et al., Nano letters 2005. •10. Chithrani and et al., Nanotechnology Letters 2006. •11. Coffin JM and et al., "Principles of Retroviral Vector Design." Retroviruses NCBI Book shelf. •12. Piazzi Leonardo da Vinci. "Optical probe for detecting sers-active molecules and process for its manufacture".Standish C Hartman and Richard C Mulligan. “Two dominant-acting selectable markers for gene transfer studies in mammalian cells”. Cell Biology Nov 1988.
  14. 14. REFERENCES: IMAGES. • Surface Enhanced Raman Spectroscopy. Semrok corp. • Chad Mirkin. “Coaxing Gold nanorods into orienting in nanospheres”. Northwestern University. Ist July 2009. •Coffin JM and et al., "Principles of Retroviral Vector Design." Retroviruses NCBI Book shelf. •Mathew T Drake and et al., “Epsin Binds to Clathrin by Associating Directly with the Clathrin-terminal Domain”. The Journal of Biological Chemistry , March 3 2000. •. Piazzi Leonardo da Vinci. "Optical probe for detecting sers- active molecules and process for its manufacture".
  15. 15. THANK YOU