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Biodefenseproject
A.D.N.
ARTIFICIAL DOG NOSE
Terrorism and biodefense
ARTIFICIAL DOG NOSE• Could we imagine the possibility of
detecting only one molecule?
ARTIFICIAL DOG NOSE
ARTIFICIAL DOG NOSE
• Narcotics• Explosives
ACTUAL SOLUTIONS
ACTUAL SOLUTIONS
Narcotics and explosives
ACTUAL SOLUTIONS
BACTERIA AND VIRUSES
ACTUAL SOLUTIONS• Up to 40 explosives• Most frequent narcotics • Up to 8 microorganisms• Viruses
• This requires quantities in NANOGRAMS
IDEA• MOLECULE, smallest AMOUNT • TO IDENTIFY ONE MOLECULE• BIOLOGICAL RECEPTOR
– BIOLOGICAL– NON BIOLOGICAL SUBSTANCES
ANTIBODY
Inmunology conceptClonal selection theory: MacFarlane Burnet 1957
1.Each lymphocyte bears a single type of receptor of unique specificity.2.Antigen interaction leads to lymphocyte activation.3.Daughter cells bear identical antigen specificity to the parent cell.
1010
1.RECEPTOR CONSTRUCTION
• Antibodies are a class of proteins secreted by B-Cells in response to the presence of a foreign substance or Antigen.
• Antigen: foreign substances which can combine with TCR or BCR or Ab and have the potential of inducing immune response (>10,000Dlt).
– binding interactions are: van der Waals forces, hydrophobic interactions, and H-bonds
ORGANICS RECEPTORS
• Hapten: substances which can combine with Ab, but cannot induce immune response independently. In another word, hapten only possess immunoreactivity.
• Carrier: enhance the immunogenicity of hapten, can be a protein, bacteria, cell...
ASPIRIN
ORGANICS RECEPTORS
• Hapten needs:– Covalent union
with a large protein.
– Cell’s surface (eritrocytes).
– MO’s surface.
Mechanism of Hapten-Carrier
• Hapten recognized by Ig receptor on B cell• Hapten-carrier endocytosed by B & APC• Carrier processed and presented on class II
MHC to Th cell • Activated Th & APC cell produces cytokines• Cytokines enable B cell to be activated to
produce anti-hapten antibodies
The three main types of armed effector T cell produce distinct sets of effector molecules.
THYMUS-DEPENDENT
THYMUS-INDEPENDENT
CYTOKINES
A second signal is required for B-cell activation either by thymus-dependent or by thymus-independent antigens.
B CELL
T CELL
2º INTERACTIONCYTOKINES
...or by thymus-independent antigens
APC
2º INTERACTION
HAPTEN
CARRIER
• Th, and APC cells recognize carrier, B cells recognize hapten
• Th, APC & B cells cooperate by interacting
• Interactions are class II self-MHC restricted
T Cell-B Cell Interactions(hapten-carrier effect)
CD40
Immunoglobulinreceptor
MHC II
B7B7 CD28
TCRTCRT helpercell
Antigen
1. Antigen presentation toTh cell
2. B7 expressed 3. Th cell is activated
and expresses CD40 ligand,
Cytokines secreted
CD40 ligand
Cytokine
Cytokine receptor
Bcell
Bcell
Bcell
T helpercell
4. Cytokine binds to cytokine receptor,
CD40 ligand binds to CD40
Bcell
Bcell
Bcell
5. B cell activated
6. B cells proliferate, differentiate, secrete Ig
Class II MHCAPCTh cell B
cellTh cell
Bcell
Bcell
B cell takes up and presents antigen
Th cell Th cellB
cell
Th cells are primed by antigen-presenting cell
B-T cell cooperationB cells receive signals from T cells
B cells divide
Bcell
Bcell
Bcell
Bcell
Antibody formingcell
Antibody formingcell
Antibody formingcell
Bmemory
cell
ORGANIC RECEPTORS
• Hapten needs:
ORGANIC RECEPTORS• Hapten needs:
CRYSTAL
APC
ORGANIC RECEPTORS• Hapten needs:
COCAINE HEROIN
ATROPINE HEPARINE
ORGANIC RECEPTORS
PENICILLIN
TNT
WHY NOT?TNT
ORGANIC RECEPTORS
ORGANIC RECEPTORS
Artificial-carrier
PROBABLY, THE SMALLEST INTELLIGENT CARRIER POSIBLE
ONE ANTIGENIC STRUCTURE FOR SMALL HAPTENS
THEORICAL PROCESS
CONTACT LENS
1.RECEPTOR CONSTRUCTION• MONOCLONAL ANTIBODIES
InmunizationSpecific Ag
AntibioticsAnimal serumhipoxantine
Isolation of monoclonal antibodies
Nourishing stratus
*Hammerling, o. J.; u. Hámmerling; j. F. Kearny (editores): monoclonal antibodies and t-cell hybridomas.Perspectives and tk(hnical advances. Research monog. In lmmunology vol. 3, amsterdam, elsevier/north holland biomed press, 1981
Mouse tumor
HAT medium
Ab
summary• ANTIBODIES = MOLECULAR
RECEPTORS• HAPTENS BECAME INUNOGENICS• NATURAL SELECTIONS OF
ANTIGENICS STRUCTURES• THERE IS NOT HISTOCOMPATIBILITY
PROBLEMS
• FACED WITH…. EXPLOSIVES• WITH TOXICS• WITH BACTERIA AND VIRUSES• WITH A WIDE RANGE OF MOLECULAR STRUCTURE
• QUANTITIES OF • 10 12 TIMES MORE SENSITIVE
• OF NANOGRAMS 10 -9 gr• ZEPTO O YOCTO GRAMOS 10 -23
ANTIBODIES TYPES
Biochips• Micromatrix of biologic macterial.• Massive obtention of information.• Classification• design• Clustering/data mining
Ab’s Microarrays
TYPESFluorescence image of SAM-Biotin capture membrane subsequent to antibody treatment
SAM-Membrane was incubated with FITC-labelled monoclonal anti-phosphotyrosine antibody PT-66 (Sigma, Order# F3145) (dilution 1:1000 in TBS) for 150 min followed by washings (5 times) with TBS-buffer. Fluorescence image was generated using FLA 3000 reader (Fuji, excitation at 473 nm, fluorescence filter 520 nm).
Chemoluminescence
Comparison of a microarray experiment. The red dots represent transcripts found with AB's microarray that are either absent or not detectable on Affymetrix whole genome microarrays.
CHEMOLUMINISCENCE• This micrograph of the Germanium detector shows an active area
about 30 µm wide, sandwiched between two interdigitated electrodes on top of a silicon dioxide base.
• optical micrograph of an array of GeOI photodetectors. The detectors at the top of the photograph are 100 µm wide, while those at the bottom measure just 10 µm
Ab’s Microarrays
Scanner molecular
ARTIFICIAL DOG NOSE
ARTIFICIAL DOG NOSE
ARTIFICIAL DOG NOSE• Molecular sensitivity• High specificity• Up to 360 different molecules at one
time• Independently
– Explosives– Toxics– Bacteria– viruses
ARTIFICIAL DOG NOSE
• LIMITATIONS– EXPERIMENTAL TECHNOLOGY– HIGH COST R and D– DIAGNOSIS TIME TOO LONG
“Dreams come true, without that possibility,
nature would not incite us to have them”John Updike
ORGANICS RECEPTORS