18768515 monoclonal antibody

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MONOCLONAMONOCLONA

LL

ANTIBODIESANTIBODIES

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NT IB ODI ESNT IB ODI ES

PROTIENS SYNTHESIZED INPROTIENS SYNTHESIZED IN

BLOODBLOOD

IMMUNITY IN BLOODIMMUNITY IN BLOOD

BLOOD SERUMBLOOD SERUM

HETEROGENOUSHETEROGENOUS

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WH T RE m bs??H T RE m bs??

SPECIFIC LYMPHOCYTESPECIFIC LYMPHOCYTE

SINGLE TYPE AbSINGLE TYPE Ab

SPECIFICITY AGAINST SPECIFICSPECIFICITY AGAINST SPECIFICANTIGENANTIGEN

DIFFICULT IN MAINTAININGDIFFICULT IN MAINTAINING

CANCER CELLSCANCER CELLS

IMMUNOGLOBULINSIMMUNOGLOBULINS

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Contents:Contents:DiscoveryDiscovery

mAb !er"#ymAb !er"#y

Sr$c$re "%& '$%cio%Sr$c$re "%& '$%cio%

Pro&$cio%Pro&$cio% Ty#es Ty#es

mAb ec!%o(o)ymAb ec!%o(o)y

mAb !eorymAb !eory

A##(ic"io%sA##(ic"io%s

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♣♣DISCOVERDISCOVER

 Y  Y ♣♣

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TH E M GI C BU LLETH E M GI C BU LLET

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P UL EHR LICH  UL EHR LICH

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Hu man-Mo use Hyb r!u man-Mo use Hyb r!

Ce ""se""s

GeorgesGeorges

KöhlerKöhler

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C#sar#sar

M"s$e n"s$e n

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Ne"s %a&e"s %a&

'er neer ne

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GREG WINTERGREG WINTER

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MONOCLONALMONOCLONAL

ANTIBODYANTIBODYTHERAPY THERAPY 

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one specific antigenone specific antigen

stimulating the patient'sstimulating the patient'simmune systemimmune system

e.g.radioimmunotherapye.g.radioimmunotherapy

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STRUCTURESTRUCTUREANDAND

FUNCTIONSFUNCTIONS

OFOFMAb...MAb...

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IgG 150kDaIgG 150kDa

2 diff. chains2 diff. chainsight chain kappaight chain kappa

lam!dalam!da"apain #a! #c"apain #a! #c

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Tu mor ( e""s)) )u mor ( e""s)) )

$r!%2 &$2 !reast cancer $r!%2 &$2 !reast cancer 

 ()*I%+DI$, recogni-e foreign ()*I%+DI$, recogni-e foreign ()*IG$),... ()*IG$),...

(!s are used to raise specific(!s are used to raise specificanti!odies on such surfaces ofanti!odies on such surfaces of

tumors//tumors//

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PRODUCTIONPRODUCTION

  HYBRIDOMA CELLHYBRIDOMA CELL

PRODUCTIONPRODUCTION

  RECOMBINANTRECOMBINANT

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HYBRIDOMAHYBRIDOMA

CELLCELL

  PRODUCTION  PRODUCTION

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RECOMBINANTRECOMBINANTAb PRODUCTIONAb PRODUCTION

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Repertoire cloning/phageRepertoire cloning/phage

display/yeast displaydisplay/yeast displayVirus/yeast used instead of MICEVirus/yeast used instead of MICE

Rapid cloning Ig gene segmentRapid cloning Ig gene segmentAbs having slightly diff. aminoAbs having slightly diff. amino

acid sequencesacid sequences

Ab of desired specificity areAb of desired specificity are

selectedselected

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T* e + , . on$s)))* e + , . on$s)))

specificity of (! to recogni-e (gspecificity of (! to recogni-e (g

sta!ilitysta!ilitytherapeutic efficacytherapeutic efficacy

detectiondetection

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TYPES OFTYPES OF!Abs!Abs

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MURINE MAbsMURINE MAbs

Initially &%ID+( technology asInitially &%ID+( technology as

usedused

"+%$,"+%$,

1.educed stimulation of cytotoity1.educed stimulation of cytotoity

2.ild allergic reactions2.ild allergic reactions

3.anaphylactic shocks3.anaphylactic shocks

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CHIMERIC MAbsCHIMERIC MAbs

urine 4aria!le region fusedurine 4aria!le region fused

onto human constant regiononto human constant region

Kappa light chainIgG1 hea4yKappa light chainIgG1 hea4y

chain6(nti!odies 758 humanchain6(nti!odies 758 human

Increases serum half lifeIncreases serum half life

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HUMANI"ED MAbsHUMANI"ED MAbs

Grafting murine hyper4aria!le aminoGrafting murine hyper4aria!le amino

acid domains into human anti!odiesacid domains into human anti!odies

958 human origin958 human origin

&umani-ed anti!ody:parent murine&umani-ed anti!ody:parent murine

(!(!Introducing ;*(*I+), likeIntroducing ;*(*I+), like chain<chain<

shuffling= site<specific mutagenesis= $<colishuffling= site<specific mutagenesis= $<coli

mutator strains and others...mutator strains and others...

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HUMAN MAbsHUMAN MAbsransgenic miceransgenic mice  ++

 phage display librariesphage display libraries

*ransferring human genes into murine genome.*ransferring human genes into murine genome.*he mouse is then 4accinated against the human*he mouse is then 4accinated against the human

immunoglo!ulinimmunoglo!ulin

"hage display li!raries transforms murine (!s in"hage display li!raries transforms murine (!s in4itro into fully human (!s.4itro into fully human (!s.

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MAb Te#$no%o&'...MAb Te#$no%o&'...

>&I)(>&I)(Dr. a an= unnan uni4ersityDr. a an= unnan uni4ersity

"iece of paper test"iece of paper test

unda onoclonal *echnologyunda onoclonal *echnology

>ompany>ompany

 (ID,= &epatitis %= &epatitis >= (ID,= &epatitis %= &epatitis >="rostrate >ancer= 4enereal disease"rostrate >ancer= 4enereal disease

and so on...and so on...

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!Ab!Ab

t$eo('t$eo('

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Antigen!"oreign invadersAntigen!"oreign invaders

Antibody!#atural defensesAntibody!#atural defenses

Monoclonal AntibodiesMonoclonal Antibodies

  $rotect$rotect

  diagnosediagnose

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$ure antibody prod.$ure antibody prod.

Immortal cellsImmortal cellsumor cell %ith mammalianumor cell %ith mammalian

cell fusioncell fusion& type of cell i.e. 'ybridoma& type of cell i.e. 'ybridoma

hence monoclonalhence monoclonal

Many (inds of cells)Many (inds of cells)

polyclonalpolyclonal

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C)n#e( t(e)t!entC)n#e( t(e)t!ent

%ind only to cancer cells%ind only to cancer cells,ho immunological response,ho immunological response

odificationodification

%ispecific anti!ody%ispecific anti!ody

 (D$"*<anti!ody directed en-yme (D$"*<anti!ody directed en-yme

  prodrug therapyprodrug therapy

 (D>><anti!ody dependent cell<mediated (D>><anti!ody dependent cell<mediated

  cytotoicitycytotoicity

( d th( d th