biosensors: analytical techniques
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ANALYTICAL TECHNIQUES
BIOSENSORS
MADE BY: DEEPANSH MODY
U101113FBT259
NIIT UNIVERSITY
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INTRODUCTION
A BIOSENSOR IS AN ANALYTICAL DEVICE WHICH IS USED TO DETER
PRESENCE AND CONCENTRATION OF A SPECIFIC SUBSTANCE IN A B
ANALYTE.
THEY HAVE APPLICATIONS IN DIVERSE FIELDS LIE DIAGNOSTICS! D
MONITORING! FOOD INDUSTRY AND ENVIRONMENTAL MONITORING
PROF. LELAND C CLAR "R. IS NOWN AS THE FATHER OF BIOSENSO
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Biosensor
DiBio receptor T#$%&'()*# Signal
Processing
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Bio sample
COMPONENTS OF A BIOSENSOR
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IT IS THE RECOGNITION COMPONENT. IT IS DESIGNED TO INTERACTSPECIFIC ANALYTE OF INTEREST AND PRODUCE AN EFFECT MEASUR
THE TRANSDUCER.
IT NEEDS TO BE IMMOBILIED IN THE VICINITY OF THE TRANSDUCER
OBTAIN PROPER PROBE ORIENTATION! AND INCREASE ACCESSIBILIT
TARGET ENYME.
IT IS DONE EITHER BY PHYSICAL ENTRAPMENT OR CHEMICAL ATTACH
CHEMICAL ATTACHMENT OFTEN INVOLVES COVALENT BONDING TOTRANSDUCER SURFACE BY SUITABLE REAGENTS.
IT IS TO BE NOTED THAT ONLY MINUTE QUANTITIES OF BIORECEPTOMOLECULES ARE NEEDED! AND THEY ARE USED REPEATEDLY FORMEASUREMENTS.
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A TRANSDUCER IS CAPABLE OF CONVERTING THE BIO6RECOGNITION
INTO A MEASURABLE SIGNAL.
IT ACTS AS AN INTERFACE! MEASURING THE PHYSICAL CHANGE THAT
WITH THE REACTION AT THE BIORECEPTOR! AND THEN TRANSFORM
ENERGY INTO MEASURABLE ELECTRICAL OUTPUT.
DEPENDING ON THE TYPE OF TRANSDUCER! A BIOSENSOR IS DIVIDE
FOLLOWING CATEGORIES:
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WORING OF A BIOSENSOR ANALYTE DIFFUSES FROM THE SOLUTION TO THE SURFACE OF THE
BIOSENSOR.
ANALYTE REACTS SPECIFICALLY AND EFFICIENTLY WITH THE BIORE
THIS REACTION CHANGES THE PHYSIO6CHEMICAL PROPERTIES OF
TRANSDUCER SURFACE.
THE TRANSDUCER THEN TRANSFORMS THE BIOLOGICAL SIGNAL IN
ELECTRICAL SIGNAL.
THIS SIGNAL IS PASSED ON TO THE MICROPROCESSOR! WHERE ITUNDERGOES AMPLIFICATION.
IT THEN REACHES THE DISPLAY! OR THE STORAGE DEVICE.
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BREATHROUGH 77GLUCOSE MONITORING BIOSENSORS:
THE GLUCOSE CONCENTRATION IN A BLOOD SAMPLE CAN BE MEASUDIRECTLY
GLUCOSE 8 O2 GLUCOSE OIDASEGLUCONIC ACID 8 H2O2
BIORECEPTOR: GLUCOSE OIDASE
TRANSDUCER:
1 OYGEN SENSOR6 MEASURES OYGEN CONCENTRATION
2 PH SENSOR6 MEASURES THE ACID ;GLUCONIC ACID
3 PEROIDASE SENSOR6 MEASURES H2O2CONCENTRATION
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APPLICATIONS OF BIOSENSORS
BIOSENSORS HAVE APPLICATIONS IN A VARIETY OF FIELDS! RANGIN
FOOD QUALITY TO DNA SEQUENCING. SOME OF THE APPLICATIONS
DNA SEQUENCING
MUTATION DETECTION
CRIME DETECTION
FOOD ANALYSIS
DRUG DEVELOPMENT
QUALITY CONTROL
MEDICAL DIAGNOSIS
INDUSTRIAL PROCESS CONTROLS
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DNA BASED BIOSENSORS
THE FUNDAMENTAL PRINCIPLE BEHIND DNA BIOSENSORS DEPENDS
SEQUENCE COMPLEMENTARITY AS PER CHARGAFF
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THEY ARE BASED ON THE OPTICAL TRANSDUCTION OF A BIOLOGICA
TO AN OPTICAL SIGNAL.
THE DEGREE OF TRANSDUCTION FOLLOWS BEER LAMBERT=S LAW.
OPTICAL BIOSENSORS CAN BE MINIATURIED BY USING FIBRE OPTI
CONVERT EMISSION SIGNAL INTO A DETECTABLE FLORESCENT SIGN
DNA PROBE AND TARGET HYBRIDISATION IS DETECTED BY A FLUOR
MARER! WHICH RESULTS IN TOTAL INTERNAL REFLECTION IN THE
WHICH IS MEASURED BY THE DETECTOR.
THE FLUORESCENCE MARER INTERCALATES WITH THE HELICAL STAND IN CASE OF SSDNA! MOLECULAR BEACONS ACT LIE FALSE HE
A MOLECULAR BEACON IS A SINGLE6STRANDED NUCLEIC ACID PRO
COMPRISING THREE FUNCTIONAL DOMAINS > A STEM! A LOOP! AND
FLUOROPHORE?QUENCHER PAIR
Optical DNA Biosensors
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DNA PROBE IS IMMOBILIED ONTO AN ELECTRODE ! AND A CHANGE
ELECTRICAL PARAMETERS ;EG: CURRENT! CONDUCTANCE! IMPEDAN
GENERATED BY THE HYBRIDISATION REACTION IS MEASURED.
Electrochemical DNA Biosen
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)-,/-(%'& $#* $$)*'+ * '%$ /#-@*!+) #*$) + *$#* '%$ $%' /#-'()***)#+)$ )$#*&.
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A STRETCH OF NUCLEOTIDE SEQUENCE WITH A FEW HUNDRED BAS
POSSESSES A MEASURABLE AMOUNT OF MOLECULAR WEIGHT. WHE
DNA PROBE HYBRIDISES WITH THE TARGET SEQUENCE! THERE IS A
INCREASE IN MOLECULAR WEIGHT! WHICH FURTHER LEADS TO A DIN THE RESONANT FREQUENCY OF THE PIEOELECTRIC BIOSENSOR
Piezoelectric DNA Biosens
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SIGNAL PROCESSING THE TRANSDUCED SIGNAL NEEDS TO BE PROCESSED BEFORE IT IS
MONITORED.
THIS IS DONE WITH THE HELP OF DNA6FET ;DNA FIELD EFFECT TRA
IT LEADS TO AN AMPLIFICATION IN THE SIGNAL PRODUCED.
HYBRIDISATION OF STRANDS LEADS TO CURRENT TRANSPORT THR
TRANSDUCER! WHICH IS FED INTO THE GATE TERMINAL OF THE FIE
TRANSISTOR.
THE NEGATIVE CHARGES RELEASED VARY THE GATE POTENTIAL! LE
A VARIED RESPONSE.
THE PROCESSING OF SIGNAL IS DONE USING THE FOLLOWING PRO
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THE SOURCE TERMINAL IS GROUNDED! AND OUTPUT IS OBTAINED AT
DRAIN TERMINAL.
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DNA SEQUENCING
IT IS DONE SO AS TO DETERMINE THE CORRECT ORDER OF BASES IN THE D
SPECIES. IT IS DONE USING PHOTOCLEAVABLE FLUORESCENT NUCLEOTIDES.
IT FOLLOWS SEQUENCING6BY6SYNTHESIS ;SBS METHOD.
THE BIORECEPTOR HERE IS THE SSDNA SEQUENCE WHICH HAS TO BE SEQ
DIFFERENT PHOTOCLEAVABLE FLUORESCENT NUCLEOTIDE ANALOGUES W
DGTP6PC6BODIPY6FL6510DTTP6PC6RG!
DATP6PC6RO!
DCTP6PC6BODIPY650
FLUOROPHORE WAS ATTACHED TO THE 5 POSITION OF THE PYRIMIDINES ANDPOSITION OF THE PURINES.
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AFTER IMMOBILISATION OF PROBE! PCR WAS CARRIED OUT ONE BASE AT
PCR BEGINS AT THE FIRST NUCLEOTIDE OF THE PROBE STRAND AND GR
COMPLEMENTARY BASE! TO WHICH ONLY ONE OF THE FLUOROPHORES
THE PCR COULD BE TERMINATED BY THE PHOTOCLEAVABLE
26NITROBENYL LINER ATTACHED TO EACH FLUOROPHORE.
IT WAS DONE USING THE UV NITROGEN LASER WITH DISTINCT
WAVELENGTHS! WHICH FACILITATED EACH FLUOROPHORE TO EMIT
LIGHT! WHICH WAS THEN DETECTED BY THE SCANNER.
MEANWHILE! THE NET STEP OF PCR WAS CARRIED BY THE
AIDO6LABELED PRIMER! WHICH IS A SELF PRIMING COMPLE ATTACHED TIMMOBILIED DNA TEMPLATE BY ENYMATIC LIGATION.
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MUTATION DETECTION
THE POINT MUTATIONS IN GENES RESULT IN VARIOUS DISORDERS!
INCLUDING CANCERS.
THE SNP CAN BE DETECTED USING PIEOELECTRIC DNA BIOSENSO
AS THE HYBRIDISATION OF TARGET DNA STARTS! SIGNALS ARE
TRANSFERRED TO QUART CRYSTAL! WHOSE RESONANT FREQUENCHANGES ACCEDING TO THE RECEIVED SIGNAL. DURING THIS PRO
THERE IS A MISMATCH BETWEEN THE PROBE=S SEQUENCE AND TH
SEQUENCE! THERE IS A DIFFERENCE IN THE RESONANT FREQUENC
QUART CRYSTAL! AND HENCE! MUTATION IS DETECTED.
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REFERENCES
WEBSITE LINS:
HTTP:??WWW.DOCSTOC.COM?DOCS?1355?PPT666BIOSENSORS6DNAPROTEIN6BIOSENSORS
HTTP:??EN.WIIPEDIA.ORG?WII?DNAFIELD6EFFECTTRANSISTOR
HTTP:??WWW.IUE.TUWIEN.AC.AT?PHD?WINDBACHER?NODE.HTML
HTTP:??WWW.ACADEMIA.EDU?1155?PRINCIPLEANDREVIEWONDNSOR
HTTP:??WWW.RESEARCHGATE.NET?POST?HOWDOESETHIDIUMBROMBRINTERCALATEWITHTHESINGLESTRANDEDDNAORRNA
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RESEARCH PAPERS:
OPTICAL AND ELECTROCHEMICAL DNA NANOBIOSENSORS!
HTTP:??WWW.MOLECULAR6BEACONS.ORG?DOWNLOAD?LI!AS01;111
RECENT ADVANCES IN FIBER6OPTIC DNA BIOSENSORS YI6MING WANIAO6FENG PANG1 ! YU6YU HANG2
FOUR6COLOR DNA SEQUENCING BY SYNTHESIS ON A CHIP USING PHCLEAVABLE FLUORESCENT NUCLEOTIDES
OPTICAL! ELECTROCHEMICAL! AND MAGNETIC DNA BIOSENSORS > AOVERVIEW! NORTHERN ILLINOIS UNIVERSITY
NUCLEIC ACID BASED BIOSENSORS FOR CLINICAL APPLICATIONS UT
BORA1!2! ARGHYA SETT1 AND DEEPIA SINGH1
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THANK YOU