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Instrumentation of Affinity Chromatography - PDF Version 1

Chromatography uses a solid support with covalently bound ligands to separate and purify target molecules. The solid support should be insoluble, chemically and mechanically stable, and easily regenerated. The ligand should strongly and specifically bind the target molecule. Affinity chromatography uses the specific interaction between a ligand and target molecule to separate it from other compounds. Various methods are used to selectively elute the bound target from the ligand to purify it.

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Rakibul Karim 71
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972 views2 pages

Instrumentation of Affinity Chromatography - PDF Version 1

Chromatography uses a solid support with covalently bound ligands to separate and purify target molecules. The solid support should be insoluble, chemically and mechanically stable, and easily regenerated. The ligand should strongly and specifically bind the target molecule. Affinity chromatography uses the specific interaction between a ligand and target molecule to separate it from other compounds. Various methods are used to selectively elute the bound target from the ligand to purify it.

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Rakibul Karim 71
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umunta Mahods d Analysis Unit-5 PToIT

pmpenens d Aiy Cmomaloqraphy Dr. Parlanya Shukla

Chromabgrahbic mada- (Solid Support) Dr. .P.3ngh Closes

Chromatoqraphic mudio or solid Aupbot or mahx is 0


Substonu to which o speuic gond is Covalintly baund.

To orm o maix oechvs ik ahauld hovu turtain


chra cterishs -
shoudd bi inAolub.u in solvent and bullas Ad
in tK rO

(i)
ahadd b1
should bi
chamically and muchanicaly stablh
oasiy 'couhlsd
to o
gond or Abau
arm on to which Jigond can bs atohaud
dv) 4shauld hove good ou propaTkas and havs
Aulabunlylora dundau oraa to atach mant.

)mmobiunad Ligond
Th Alachon d igand is basad on natuu d macromobul
o b ablafad.

ihun 0 ho monu uupto1 protain is to bs purifizd by


ainity chromaBbgrabhy , Ki hormon iBAf ion
Idia Condidata or Ri ligand.
tor onibody isolabm, an antqen may br uwkd as baand.
tor

o n en2ymu to br puridiud a Aubsrakh onalog, inhibitou


Coloctor oi alatbr may bt uud as gond
OVETlaping d tageh masuwl Shacr OTm enAuT Rat Ru torget
and garcd
moluul has ull ortuu to Ki ligand.
TorgeB molu ule

igond

Brad or mahrix
TnAtamnolen andphodo.bgy Atiniy Chromalogrebby Dr. Parjanya Shukla
&
xTnaumant or ComponanB, oainity chromabgrobhy Dr. N. P. 8tngh Cluascoa

Ainihy Subport (Mahix)


SpatT Arm
Ligands
Looding Target
Saparahan lwhon
Compourd
C a

COO

C
o

''a lol
Immobliged OO
ligand
Puxipud tompaund

AHinity mpdium is 2qilibrobd wih bulr

Somplu i abbliud und condikon Ke ovOY Ahaüic


binding taage molicul to te gand and
unbobnd matYia washu rough column.
Torget Aubtanu bind spsuically bud uvevaibly to ligond)

Torge moluuuu is ucovud by changing cmdiionA


whichovo olubom dbund moli ulusb

CJukion i rfomad b wing,a tompetikiva mabuls uih


igond (Spriically)
bchanging ph, ionic ahrangh or polaviy (Ron- Pralial)
TarqeB moluulus ia colluctad in a punijiud, cananated om.
Ainity mudium u-aquilibroked wih binding buar

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