Qhov sib txawv tseem ceeb ntawm gel pom thiab affinity chromatography yog tias gel pom chromatography nyob ntawm qhov sib txawv ntawm qhov hnyav molecular lossis qhov loj ntawm cov qauv tshuaj ntsuam xyuas, qhov affinity chromatography nyob ntawm qhov affinity ntawm analyte rau ib qho immobilized ligand.
Lub sij hawm chromatography hauv analytical chemistry yog hais txog cov txheej txheem ntawm kev sib cais ntawm cov khoom hauv ib qho kev sib xyaw uas siv cov txheej txheem sib txawv. Chromatography yog ib lub npe sib sau ua ke uas sawv cev rau ntau yam kev sib txawv rau kev sib cais.
Gel Filtration Chromatography yog dab tsi?
Gel filtration chromatography yog cov txheej txheem tshuaj ntsuam xyuas uas kev sib cais ntawm cov khoom nyob ntawm qhov sib txawv ntawm qhov hnyav lossis qhov loj me. Cov txheej txheem no tseem hu ua qhov loj-exclusion chromatography lossis molecular-sieve chromatography. Cov txheej txheem kev sib cais hauv cov txheej txheem no nyob ntawm qhov sib txawv ntawm qhov muaj peev xwm ntawm cov qauv hauv cov qauv kom nkag mus rau hauv qhov hws ntawm gel-filtration nruab nrab.
Nyob rau hauv cov txheej txheem gel filtration, cov theem nyob ruaj ruaj muaj cov hlaws dai ntawm cov hydrated, daim txhuam cev zoo li cov khoom uas muaj qhov pores ntawm molecular qhov ntev uas muaj qhov nqaim ntawm qhov ntau thiab tsawg. Yog tias peb dhau cov tshuaj aqueous uas muaj cov molecules ntawm ntau qhov ntau thiab tsawg los ntawm ib kem uas muaj cov "molecular sieves", cov molecules uas loj dua qhov pores ntawm qhov nruab nrab pom txav mus los ntawm kab. Nyob rau hauv sib piv, cov me me molecules nkag mus rau hauv lub qhov hws ntawm cov gel thiab zoo li maj mam txav mus rau sab. Molecules elute los ntawm kem kom txo qis molecular loj. Ntawm no, qhov txwv tsis pub ntawm cov gel yog cov molecular loj ntawm cov molecules me tshaj plaws tsis tuaj yeem nkag mus rau lub qhov hws ntawm cov gel.
Muaj ntau hom kev gel filtration, xws li indirect gel filtration method, steady-state gel filtration, gel bead dialysis, etc. Indirect gel filtration chromatography yog pab tau rau kev txiav txim siab ntawm cov thyroid hormones dawb. Steady-state gel filtration chromatography yog ib qho kev qhia tsis ncaj uas feem ntau siv rau kev ntsuas cov tshuaj dawb steroids xws li cortisol, testosterone, thiab lwm yam. Gel hlaws dialysis, ntawm qhov tod tes, yog ib qho kev hloov kho ntawm khov kho-state gel filtration uas yog kev sib piv yooj yim dua.
Affinity Chromatography yog dab tsi?
Affinity chromatography yog cov txheej txheem kev tshuaj xyuas thiab kev sib cais uas nyob ntawm ib qho kev sib txuas lus tshwj xeeb ntawm ib qho immobilized ligand thiab nws tus khub khi. Qee qhov piv txwv suav nrog cov tshuaj tiv thaiv kab mob-antigen binding, enzyme-substrate khi, thiab enzyme-inhibitor khi. Yog li ntawd, cov txheej txheem no muaj ntau yam kev siv tseem ceeb hauv nucleic acid purification, protein purification los ntawm cell extracts, thiab purification los ntawm cov ntshav.
Hauv cov txheej txheem no, cov cuab yeej tseem ceeb tshaj plaws yog ligand immobilization. Peb tuaj yeem siv ntau yam ntaub ntawv xws li acrylates thiab silica gel rau qhov no. Nws yog ib qho tseem ceeb los tiv thaiv steric cuam tshuam ntawm lub hom phiaj molecule rau ligand. Ntxiv mus, ib qho inhibitor txuas nrog rau theem khoom. Peb hu qhov no inhibitor ib tug spacer. Classically, ib tug spacer muaj ib tug hydrocarbon saw.
Lub sijhawm nyob ruaj khov ntawm qhov kev sib raug zoo chromatography muaj cov tub ntxhais, spacer, thiab ligand. Qee lub sij hawm, nws kuj muaj cov hlau ion uas txuas nrog lub ligand. Qhov kev nyiam tshaj plaws rau theem nyob ruaj khov hauv cov txheej txheem no yog agarose gel vim tias nws yooj yim dispensed los sau thiab ntim cov kem nrog resin txaj ntawm txhua qhov me me, thiab nws loj txaus rau biomolecules ntws mus rau hauv thiab los ntawm cov hlaws dai. Cov ligands tuaj yeem txuas rau cov hlaws polymer hauv ntau txoj hauv kev. Cov tshuaj sib xyaw ua ke ntau tshaj plaws yog cyanogen bromide, epoxide, epoxide nrog C6 acid, thiab diamin. Ntawm qhov tod tes, cov ligand peb tuaj yeem siv sib txawv raws li lub hom phiaj, xws li cov tshuaj tiv thaiv-antigen, hlau lossis txhuas ions-phosphoproteins, avidin-biotin, glutathione-GST, chelator-His-tagged proteins, thiab lwm yam.
Qhov txawv ntawm Gel Lim thiab Affinity Chromatography yog dab tsi?
Chromatography yog ib qho tseem ceeb analytical txheej txheem. Gel filtration chromatography thiab affinity chromatography yog ob qho kev hloov pauv tseem ceeb ntawm chromatography. Qhov sib txawv tseem ceeb ntawm gel filtration thiab affinity chromatography yog tias gel filtration chromatography nyob ntawm qhov sib txawv ntawm qhov hnyav molecular lossis qhov loj ntawm cov qauv tshuaj ntsuam xyuas, qhov affinity chromatography nyob ntawm qhov affinity ntawm ib qho tshuaj ntsuam xyuas rau ib qho immobilized ligand.
Summary – Gel Lim vs Affinity Chromatography
Muaj ntau hom kev kawm chromatographic raws li lawv daim ntawv thov thiab qhov xwm txheej ntawm tus qauv ntsuas. Gel filtration thiab affinity chromatography yog ob hom kev kawm. Qhov sib txawv tseem ceeb ntawm gel filtration thiab affinity chromatography yog tias gel pom chromatography nyob ntawm qhov sib txawv ntawm qhov hnyav molecular lossis qhov loj ntawm cov qauv tshuaj ntsuam xyuas, qhov affinity chromatography nyob ntawm qhov affinity ntawm ib qho tshuaj ntsuam xyuas rau ib qho immobilized ligand.
