3/26/2017 ProductionofRecombinantHumanGrowthHormoneSomatotropin

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ProductionofRecombinantHumanGrowthHormoneSomatotropin
BY:NidhiUppangala|Category:Biotechnologyproducts|Submitted:2010082710:43:10

Article Summary: "Recombinant human growth

hormone is generally produced by inserting the
human growth hormone gene into plasmids of
E.Coli bacteria. Recombinant bacterial cells are
cultured and human growth hormones produced by
these bacteria are extracted from the
extracellular media..."

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Introduction:

RecombinantDNAtechnologyiswidelyusedintheproductionoftherapeuticagentssuchas
hormones,cytokines,growthfactors,antibiotics,vaccines,bloodproductslikealbumin,
thrombolytics,fibrynolytics,clottingfactorssuchasfactorVII,factorIX,tissueplasminogen
activatorandmanymore.Allthesetherapeuticagentscanbeproducedinalargequantityandalso
moreeconomicallybyusingrecombinantDNAtechnology.

Growthhormonesareproducedinthepitutarygland,andareinvolvedintheoverallgrowthofthe
bodybyincreasingtheproductionofprotein,thusincreasetheaminoaciduptakebythebodycells.
Growthhormonealsoincreasestheuseoffatasbodyfuel.

RecombinanthumangrowthhormoneSomatotropinProduction:

Recombinanthumangrowthhormoneisgenerallyproducedbyinsertingthehumangrowthhormone
geneintoplasmidsofE.Colibacteria.Recombinantbacterialcellsareculturedandhumangrowth
hormonesproducedbythesebacteriaareextractedfromtheextracellularmedia.

Humangrowthhormonesarealsoproducedandextractedusinganimalcellculturearenowadays
areusedintreatingpatientswithrenalcarcinomaandalsototreatchildrenwhoaresufferingfrom
humangrowthdeficiency
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3/26/2017 ProductionofRecombinantHumanGrowthHormoneSomatotropin

Limitations:

Duringtheproductionofthehumangrowthhormoneinthebodyitisattachedwithsignalpeptide
with26aminoacids.Thissignalpeptideiscleavedinthebodybeforereleasingthehumangrowth
hormonetodotheirbiologicalfunction.

Duringtheproductionofrecombinanthumangrowthhormonethesignalpeptidesinterruptstheir
production.FirstcDNAisproducedusingmRNAencodingforhumangrowthhormone.ThencDNA
areintegratedintotheplasmidandareinsertedintotheE.Colibacteria,thentheyareculturedand
humangrowthhormoneareextractedalongwiththesignalpeptidechain.AsE.Colibacteriadoes
nothavethemechanismtocleavesignalpeptidechain.


NewApproach:

1.Toresolvetheinterruptionofsignalpeptidesintheproductionofhumangrowthhormone,the
basesequencecodingforsignalpeptidesofaround26aminoacidsalongwithneighbouring24
aminoacidsiscutinthecodingcDNAusingrestrictionenzymeEcoR1.

2.Deleted24aminoacidsequenceofhumangrowthhormonecDNAisfreshlypreparedandligated
totheremaininghumangrowthhormonecDNA.

3.ThiscodingcDNAisthenintegratedintotheplasmid,theninsertedintoE.Colibacterialcell

4.Theserecombinantbacterialcellsarecultured,andthenhumangrowthhormonesareextracted
fromthemedia.

ProductionofRecombinantHumanGrowthHormoneinChineseHamsterOvary(CHO)Cells:

Thisapproachhasgotsomeadvantagessuchasrecombinanthumangrowthhormoneissecreted
directlyintotheproteinfreemedia.Hencehormonecanbeextractedandpurifiedusingsimple
methods,andthisalsoavoidsresolubilizationofinclusionbodiesandrefoldingofproteins.

ExtractionofSomatotropin:

Somatotropinisagrowthhormonecontains191aminoacidsandproducedbytheanteriorpitutary
orinotherwordsfrontsectionofthepitutarygland.Thishormoneisactivatedbyanotherhormone
producedintheliverknownassomatomedin.Therebysomatotropincausesgrowth.

2.Inthecaseofhumangrowthhormonesomatotropin,theproteinissequesteredinrefractile
bodieswithinthecytoplasmofthebodycells.

3.Refractilebodiescanberecoveredfromthecellculturebydisruptingthecelltoreleasethe
refractilebodies.Theserefractilebodiesarethencollectedaspelletbyusingdifferential
centrifugationmethod.

4.Theserefractilebodiesaresolubilizedusingsuitablechotropicagents,suchasureaorguanidine
hydrochlorideatpH912oralkalinepH.

5.Solubilizedproteinsarethentreatedwithoxidizingagent,andthisformsintramoleculardisulfide
bondsandproteinisrefoldedtoformitsbiologicallyactivestructure.

6.Biologicallyactivesomatotropinsareusedtoenhancethegrowthofanimals,andalsoanimal
productivitybyadministeringthesehormonesbysubcutaneousinjectionorintramuscularinjection.
Theycanalsobeadmisteredasimplants.

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Uses:

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3/26/2017 ProductionofRecombinantHumanGrowthHormoneSomatotropin

SomatotropinisalsousedtotreatpatientswithTurner'ssyndromeandalsoinchronicrenal
insufficiency.

8.Bovinesomatotropinisusedeffectivelytoincreasemilkproductioninlactatingcow.

9.Porcinesomatotropinisusedeffectivelytoimprovefeedefficiencyandtoincreasemeatcontent,
byadministeringittofinishinghogs.

Someoftheexamplesofrecombinanthumangrowthhormoneare

ProtropinproducedbyGenetechCompany

HumatropeproducedormanufacturedbyEriLillyCompany

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Commentsonthisarticle:(3commentssofar)

CommentBy Comment
nairutipatel
goodone...theinformationhelpedmealot.
2011080803:27:30206
sadhana tothepointandprecise...indeedaverygoodarticle....proved
2012051903:35:09293 beneficial..
Dr.Mohamadpanah
Thanksalot
2012111922:46:47356

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