Hepatitis B Vaccine production.ppt

For subject #Ind. Micro# SHC students

1. Large scale production of
recombinant molecules
HBV vaccines
By
Mr. K.VIJAY M.Sc., TNSET, NET(LS), (Ph.D.)
Assistant Professor,
Department of Microbiology,
Sacred Heart College,
Tirupattur,TN,India.

2. •HepatitisBisaviralinfectionwhichattacksthecellsoftheliver.It
canbeveryserious,causingchronicliverfailure,livercancerand
death
•HepatitisBcanbepreventedbyavaccination,andincountries
likeIndiawhereitisrelativelycommon.Acourseofthree(3)
vaccineinjectionsaregivenwiththesecondinjectionatleastone
monthafterthefirstdoseandthethirdinjectiongivensixmonths
afterthefirstdose.
•AfterwardanimmunesystemantibodytoHBsAgisestablishedin
thebloodstream.Theantibodyisknownasanti-HBsAg.This
antibodyandimmunesystemmemorythenprovideimmunityto
hepatitisBinfection.Thefirstvaccinebecameavailablein1981.
•HepatitisBvaccineisdevelopedforthepreventionofhepatitis
Bvirusinfection.Thevaccinecontainsoneoftheviralenvelope
proteins,hepatitisBsurfaceantigen(HBsAg).Itisproducedby
yeastcells,intowhichthegeneticcodeforHBsAghasbeen
inserted.

3. Recombinant Hepatitis B Vaccine
(HBsAg)
•Oneofthemostrecentdevelopmentsistheproductionofa
vaccineagainsthepatitisBusinggeneticallymodifiedyeast
cells.
•TheGenecodingfortheHBsAgisisolatedandclonedintoa
VectorUnderthecontrolofastrongpromoter.
•TheclonedgeneistransferredtotheYeastExpression
system.
•Thegeneisallowedtoexpressintheyeastandthe
recombinantproteinproductoftheHepatitisisobtained.
•Theproteinislaterpurifiedandusedforvaccination.

4. Geneencodingthe226aminoacidhepatitisBsurfaceantigen
(HBsAg),wasclonedintoyeast,Saccharomycescerevisiae.
The5'endoftheHBsAggenewasreplacedwithanotherDNA
segmentsoastooptimizesynthesisinyeast.
High-cell-densityfermentationsoflaboratorystrainsofyeasthave
beendevelopedfortheproductionofHBsAg.
TheHBsAg(lipoproteinparticle)incelllysateshasbeenpurified
toobtainhomogeneity.

5. ProductionofRecombinantHBsAgvaccine
1.HBsurfaceantigencodinggeneisisolatedfromtheHBvirus.
2.AplasmidDNAisextractedfromabacteriumandiscutwith
restrictionenzymeformingtheplasmidvector.
3.TheisolatedHBsurfaceantigenproducinggeneisinsertedinto
thebacterialplasmidvectoronformingtherecombinantDNA.
4.ThisrecombinantDNA,containingthetargetgene,is
introducedintoayeastcellformingtherecombinantyeastcell.
5.Therecombinantyeastcellmultipliesinthefermentationtank
andproducestheHBsurfaceantigens.
6.TheHBantigensareextracted,purifiedandbottled.Itisready
forvaccinationinhumans.