applicationsofbiotechnology-200403171418 (1).pdf

Application of biotechnology.. class 12 project............,#####№###########################################################################################################

1. APPLICATIONS OF BIOTECHNOLOGY IN
MEDICINE
Dr. Saji Mariam George
Associate Professor
Assumption College Autonomous
Changanacherry

2. APPLICATIONS OF BIOTECHNOLOGY
IN MEDICINE
1.Production of human insulin
Insulin –the hormone produced by the beta
cells of the Islets of Langerhansof Pancreas
A protein –2 polypeptides
A chain -21 amino acids
B chain -30 amino acids
Used for the treatment of Diabetes -
Hyperglycemia

3. Human Insulin
•Humulin–first therapeutic product produced by rDNA
technology (Eli Lilly & Co. , USA)
•Recosulin(ShreyaLife Science, Pune)
•Humanzinsulin(Sarabhai -Ahmedabad)
•Lentard40 (Ahmedabad)
•Human Insulatard(Novo Nordisk, Mumbai, Maharashtra)
•Huminsulin–R (Ranbaxy Ltd. Haryana)

4. •Thegeneforhumaninsulinisinsertedinto
E.colibyrDNAtechnology.
•ThegeneforAchainandBchainare
introducedintoseparateplasmids(pBRtype
vector)–therecombinantplasmidsare
introducedintoE.colibytransformation.
•CultureoftransformedE.colihavinghuman
insulingeneinbioreactorsandharvesting.

6. 2.Productionofhumangrowthhormone(hGH)
hGH–Somatotropin–thesecondgeneticallyengineered
pharmaceuticalapprovedforhumanuse(1985,U.SFood
andDrugAdministration)
Asinglepolypeptidechainof191aminoacids.
ThehGHcodingsequencewasinsertedintoaplasmid
carryingthelacregulatorysignalsandintroducedinto
E.colibytransformation.ThehGHsynthesizedinE.coliwas
foundtobeactiveinhumans.
Somatotropinisrequiredfornormalgrowth
Earliersource–humancadavers

7. Production of human growth hormone (hGH)

8. 3. Production of Vaccines
Vaccination
Aneffective,safestandmostpowerfultool
ofmedicinetopreventsuffering,disability,
anddeathfromvariousinfectiousdiseases.
Reducethemortalityandmorbiditydueto
severalinfectiousdiseases–Tetanus,
Measles,Mumps,Poliomyelitisetc.

9. Vaccines –(L. vacca= cow) may consists of
i.Live attenuated microbes(Attenuate –to reduce
the virulence of a bacterium or virus to make a
vaccine
ii.Killed, inactivated microbes
iii.Purified microbial components
iv.Polysaccharide –carrier protein conjugates
v.Recombinant proteins

10. •EdwardJenner(1796)–
developedfirstvaccine-
usedcowpoxvirusto
vaccinateagainsthuman
smallpox(causedby
Variolavirus).
2. Small pox
1. Cow pox

11. •Louis Pasteur(1885)-
coinedthetermvaccine
•Developedvaccineagainst
rabies
•Geneticallyengineered
Vacciniavirus(Cowpox
virus)capableof
synthesizingantirabies
vaccinewasdeveloped
in1984 (Wistar
Institute,Philadelphia)

12. HepatitisBvaccine–thefirstrecombinant
vaccineforhumanuse(1984),against
HepatitisBvirus(HBV).
•GeneticallyengineeredTobaccoplantscan
beusedfortheproductionofrecombinant
vaccinesagainstHepatitisBvirus–HBsAg
gene(HepatitisBsurfaceantigengene)may
beintegratedintotheplantgenomeandthe
recombinantproteinproducedbytheplantis
purifiedforuseasavaccine.

13. VLP–Virus Like Particles Parenteralvaccine -administered or occurring
elsewhere in the body than the mouth and alimentary canal.

14. Production of Hepatitis B vaccine
•Producedbycloning
HBsAggene(Hepatitis
Bsurfaceantigen
gene)ofthevirusin
yeastcells.

15. 4. Gene therapy.
Potentialtoolfortreatinginheritedhumandiseases-
itallowsthetransferofgeneticinformationinto
patienttissuesandorgans.
Genetherapyisstillhighlyexperimental,buthasthe
potentialtobecomeanimportanttreatment
regimen.
Involvesaddinganormal(wild–type)copyofagene
tothegenomeofanindividualcarryingdefective
copiesofthegene.
Ifsuccessful,thetransgene(fortransferredgene)will
synthesizethemissinggeneproductandrestorethe
normalphenotype.

16. Gene therapy -Types.
1.Somatic –cell gene therapy(Non heritable
gene therapy)
Treatthediseasesymptomsofthe
individual,butwillnotcurethedisease.
Thedefectivegene/swillstillbepresentin
thegerm-linecellsofthepatientafter
somatic–cellgenetherapyandmaybe
transmittedtothechildren.
Allgenetherapytreatmentsforhuman
diseasesaresomatic-cellgenetherapies.

17. Addfunctionalcopiesofthegenethatis
defective–donotreplacethedefectivegene.
Wildtypegenesareintroducedintothecells
homozygousorhemizygousforamutant
alleleusingretroviruses(RNAvirusesthat
replicatesusingreversetranscriptase)or
Adenovirusesandgetthegeneexpressed.

18. Adenoviruses–thevirusesthat
attackthemucosaofthe
upperrespiratorytractand
lymphnodes.
• Most commonly
employed vectorfor
cancergenetherapy-canbe
replication-defective;some
essentialviralgenesare
removedwhichinturnare
replacedbyagenecassette
-atypeofmobilegenetic
element-canmove
between different
constructs-containagene
andarecombinationsite–
expressesa foreign
therapeuticgene.
Image:https://www.slideteam.net/0914-

19. Gene therapy using an adenovirus vector
Image: https://courses.lumenlearning.com/

20. •Somatic–cellgenetherapy-Pioneeringstudies
onthecorrectionofadenosinedeaminase
deficiency,alymphocyte-associatedsevere
combinedimmunodeficiency(SCID).
•Successfullycarriedoutinthetreatmentof
Adenosinedeaminase-deficientseverecombined
immunodeficiencydisease(ADA¯SCID),arare
autosomaldiseaseoftheimmunesystem,in
1990.Theenzymeadenosinedeaminaseis
encodedbyageneonchromosome20.Children
withSCIDwereoftenknownas'bubblebabies'.

21. SCID
“DavidVetterfromTexashad
SCIDandhadtoliveina
sterileenvironmentfor
mostofhislifeduringthe
1970/80s.Hewasknown
tothemediaas'theboyin
the plasticbubble'
andworeaspecial
'spacesuit'toprotecthim
frominfections.”
https://www.yourgenome.org
/
Image credit:NASA Johnson
Space Center

22. Somatic cell gene therapy for SCID
STEPS
1)Isolation of White blood corpuscles (WBCs )from the patient.
2)Introduction of functional copies of the ADA gene into these
cells.
3)Demonstration of transgeneexpression in these cells.
4)Infusion of the transgenic cells back into the patient.
EffectshortlivedbecauseofthelimitedlifespanoftheWBCs
–repeatedinfusionsofWBCscarryingfunctionalADAgenesare
necessary.

23. Somatic cell gene therapy for SCID

24. Stem cells as vehicles for gene therapy
FirstusedtotreattwoinfantswithAdenosine
deaminase-deficient severe combined
immunodeficiencydisease(ADA¯SCID,1993).
•BonemarrowstemcellsthatgivesrisetoWBCscould
beusedtotreatADA¯SCIDwhichcouldprovidea
permanentorlong-termtreatmentofthedisease.
•In2000,BritishandFrenchPhysiciansusedSomatic
cellgenetherapyforthetreatmentofX-linkedSCID
inboys.(causedbythemutationsinageneonX
chromosome–afatalinheriteddisease–individuals
havenofunctionalimmunesystem).

25. 2.Germ-linegenetherapy(Heritablegene
therapy)-modifyingthegenesineggor
spermcells–passthegeneticchangesto
futuregenerations.
Performedonmiceandotheranimals,noton
humans–moralandethicalconsiderationsare
involvedinanydecisiontoperformgerm–line
modificationsofhumans.

26. Gene replacements ( Targeted gene
replacements)
•Theidealgenetherapy–involvesthe
replacementofthedefectivegenewitha
functionalgene.
•Researchersaretryingforsuchgene
replacements(targetedgenereplacements).

27. 5.Production of Monoclonal antibodies
Antibodies(Immunoglobulins)–proteins
synthesizedinbloodagainstspecificantigens.
Heterogeneous–containamixtureof
antibodies(polyclonalantibodies)–donot
havespecificity.
Aspecificlymphocyte,afterisolationand
cultureinvitro,becomescapableofproducing
asingletypeofantibodywhichhasspecificity
againstaspecificantigen–monoclonal
antibodies–usedinthediagnosisofdiseases.

28. Hybridoma techniqueforthecontinuous
productionofantibodies-(GeorgeKohlerand
CaeserMilstein,1974–NobelPrize1984in
Physiology&Medicine).
Hybridoma-formedbythefusionofamyeloma
cell(bonemarrowtumourcell)andanantibody
producinglymphocyte.
KohlerandMilstein–isolatedclonesofcellsfrom
thefusionoftwoparentalcelllines–
lymphocytesfromspleenofmiceimmunized
withsheepRBCsandmyelomaandmaintained
invitro–producedantibodieswhichimmunized
myelomacells.

29. Hybridoma-maintainthecharacterof
lymphocytestosecreteantibodiesand
ofmyelomacellstomultiplyinculture–
capableofcontinuousproductionof
antibodies.

30. Hybridomatechnique for the continuous
production of antibodies
https://www.biomol.com/

31. Hybridomacells grown in tissue culture
Image:https://commons.wikimedia.org/

32. Applications of Monoclonal Antibodies
Usefulinthediagnosisofdiseases–Cancer,
Allergy,Viraldiseases-e.g.ForAIDSdetection–
ELISA(EnzymeLinkedImmunosorbentAssay).
ForclassificationofABObloodgroups,Rhetc.
ForthetreatmentofCancer-antibodiesspecificto
acancercellcanbelinkedwithatoxin
polypeptidetoyieldaconjugatemolecule,
immunotoxin–theantibodywillensureits
bindingspecificallyandonlytothetargetcellsand
theattachedtoxinwillkillsuchcells.
Usefulinthepurificationofantigensspecificto
pathogens,whichcanbeusedasvaccines.
ABZYMES –ANTIBODIES THAT CAN FUNCTION AS
ENZYMES

33. 6.Biopharming(Molecular farming or Molecular
pharming)
•Genesthatencodethebiosynthesisofcertain
proteinsormetabolicproducts(Secondary
metabolites)whicharepharmaceutically
importantareinsertedintosuitablehostsby
recombinantDNAtechnology(rDNAtechnology)
–getthemexpressedandtheproductsare
harvestedandusedtoproducepharmaceuticals.
•Transgenicproteinsaremostcommonly
producedbytransgenicbacteriaoryeastina
bioreactor.

34. i)Biopharmingin plants
Transgenicplantsoranimalsare
usedasbioreactorstoproduce
therapeuticproteins,drugs,
vaccinesetc.–Biologics.
E.g. Transgenic tobacco, Potato,
Duck weed(Lemnaminor)
Geneticallymodifyingplantsso
thattheycanproducecertain
proteinsandothercomponents
likesecondarymetabolites
whicharetherapeutically
important.
Duck weed
(Lemnaminor)

35. The first recombinant plant –derived protein
(PDP) was human serum albumin produced in
transgenic tobacco and potato plants.
Transgenicbanana,tomato,potato-to
produceediblevaccinesagainstcertain
diseases(e.g.HepatitisB)
Transgenic tobacco –to produce Cholera
vaccine (ChlorogenInc.)

36. Plant Made Pharmaceuticals(PMPs)
Advantages
Low cost of production
Large production capacity
Safer than recombinant proteins from
microorganisms
Do not carry potentially harmful human or animal
viruses into the drug.
Theseedsandfruitsofplantsprovidesterile
packagingcontainersforthevaluabletherapeutics
andguaranteeacertainstoragelife.

37. Plant Made Pharmaceuticals(PMPs)-
Disadvantages
•Concernabout the safety of GM foods –
edible vaccines –for human consumption.
•GM plants may cross pollinate with non-GM
crops.

38. Biopharmingin animals
Creation of transgenic mammals(Cow, Sheep, Goat )
that can produce the biopharmaceutical in its milk.
Thefirstdrugproducedingeneticallymodified
livestockisATryn-Antithrombinprotein(brandnameof
theanticoagulantantithrombinmanufacturedbythe
Massachusetts-basedU.S.companyrEVOBiologics)
purifiedfromthemilkofgeneticallymodifiedgoats.
Advantages
Milk is produced in large quantity.
Purification of the biopharmaceutical from milk is easy.

39. THANK YOU