Qhov sib txawv tseem ceeb ntawm affinity thiab ion txauv chromatography yog tias peb tuaj yeem siv affinity chromatography cais cov khoom siv los yog cov khoom tsis muaj nqi nyob rau hauv ib qho sib tov whereas peb tuaj yeem siv ion txauv chromatography cais cov khoom sib xyaw ua ke.
Chromatography yog txheej txheem uas peb tuaj yeem siv los cais cov khoom xav tau hauv kev sib tov. Muaj ntau hom xws li kua chromatography, roj chromatography, thiab lwm yam. Cov affinity chromatography thiab ion exchange chromatography yog ob pawg ntawm cov kua chromatography. Tsis tas li ntawd, hauv cov txheej txheem no, muaj ob theem. Namely, lawv yog cov theem nyob ruaj ruaj thiab mobile theem. Lub hom phiaj ntawm cov txheej txheem no yog cais cov khoom, nyob ntawm qhov sib txuas ntawm cov khoom sib txuas, nyob rau hauv lub xov tooj ntawm tes mus rau saum npoo ntawm qhov chaw nres tsheb.
Affinity Chromatography yog dab tsi?
Affinity chromatography yog cov txheej txheem biochemical uas peb siv los cais cov khoom sib xyaw ua ke nyob ntawm kev sib cuam tshuam ntawm cov khoom no.
Kev sib cuam tshuam uas peb siv hauv qhov no suav nrog cov hauv qab no:
- Antigen-antibody interactions
- Enzyme-substrate kev sib cuam tshuam
- Receptor-ligand interactions
- YProtein-nucleic acid sib cuam tshuam
Hauv cov txheej txheem no, peb siv cov khoom molecular ntawm molecules rau cov txheej txheem sib cais. Ntawm no, peb tso cai rau qhov xav tau kev sib cuam tshuam nrog rau theem nyob ruaj khov ntawm hydrogen bonding, kev sib cuam tshuam ionic, txuas hniav disulfide, kev sib cuam tshuam hydrophobic, thiab lwm yam. Cov molecules uas tsis cuam tshuam nrog rau theem nyob ruaj ruaj yuav elute ua ntej. Yog li, peb tuaj yeem cais nws los ntawm kev sib tov. Cov kev xav tau sib txuas yuav txuas nrog rau theem nyob ruaj khov. Yog li ntawd, peb tuaj yeem tshem tawm nws siv cov kuab tshuaj eluting thiab ua kom nws elute cais nws ib yam.
Daim duab 01: A Chromatographic Kem
Affinity chromatography yog qhov muaj txiaj ntsig zoo hauv kev ua kom huv thiab ua kom pom tseeb ntawm cov khoom sib xyaw uas siv cov tshuaj tsis haum. Tsis tas li ntawd, nws muaj txiaj ntsig zoo hauv kev txo cov khoom tsis zoo hauv kev sib tov. Thaum xav txog cov cuab yeej uas peb siv rau cov txheej txheem no, peb yuav tsum siv ib kem uas muaj peb theem nyob ruaj khov. Tom qab ntawd, peb yuav tsum thauj cov theem mobile uas muaj cov biomolecules uas peb yuav cais. Tom ntej no, cia lawv khi nrog rau theem nyob ruaj khov. Tom qab ntawd, siv lub tshuab ntxhua khaub ncaws, peb tuaj yeem cais cov biomolecules uas tsis yog lub hom phiaj, tab sis lub hom phiaj molecules yuav tsum muaj kev sib raug zoo siab rau theem nyob ruaj ruaj kom ua tiav cov txheej txheem sib cais.
Ion Exchange Chromatography yog dab tsi?
Ion chromatography yog ib hom kua chromatography uas peb tuaj yeem txheeb xyuas cov tshuaj ionic. Feem ntau, peb siv nws los tshuaj xyuas cov anions thiab cations inorganic (piv txwv li chloride thiab nitrate anions thiab potassium, sodium cations). Txawm hais tias nws tsis tshua muaj, peb tuaj yeem txheeb xyuas cov organic ions thiab. Tsis tas li ntawd, peb tuaj yeem siv cov txheej txheem no rau kev ua kom huv ntawm cov protein vim tias cov proteins raug them cov roj molecules ntawm qee qhov pH qhov tseem ceeb. Ntawm no, peb siv lub sijhawm ruaj khov uas cov khoom them tuaj yeem txuas tau. Piv txwv li, peb tuaj yeem siv cov resin polystyrene-divinylbenzene copolymer ua cov khoom txhawb nqa.
Daim duab 02: Phase of Ion Exchange Chromatography
Txhawm rau piav qhia qhov no ntxiv, theem nyob ruaj khov tau kho ions xws li sulfate anions lossis quaternary amine cations. Txhua qhov no yuav tsum koom nrog kev tawm tsam (ib qho ion nrog tus nqi sib txawv), yog tias peb yuav tsum tswj qhov nruab nrab ntawm qhov system no. Nyob rau hauv no, yog hais tias tus counterion yog ib tug cation, ces peb npe lub system raws li ib tug cation exchange resin. Tab sis, yog tias qhov counterion yog anion, lub kaw lus yog anion pauv resin.
Muaj tsib theem loj hauv kev sib pauv ion;
- pib theem
- Adsorption ntawm phiaj
- pib ntawm elution
- End of elution
- Regeneration
Qhov txawv ntawm Affinity thiab Ion Exchange Chromatography yog dab tsi?
Affinity chromatography yog cov txheej txheem biochemical uas peb siv los cais cov khoom sib xyaw ua ke nyob ntawm kev sib cuam tshuam ntawm cov khoom no thaum ion chromatography yog ib hom kua chromatography uas peb tuaj yeem txheeb xyuas cov tshuaj ionic. Yog li, qhov sib txawv loj ntawm affinity thiab ion exchange chromatography yog tias peb tuaj yeem siv ion txauv chromatography nkaus xwb rau kev sib cais ntawm cov khoom siv ionic thaum lub affinity chromatography muaj peev xwm sib cais ob qho tib si them thiab tsis muaj nqi. Thaum xav txog lub hauv paus ntsiab lus ntawm kev ua haujlwm, qhov sib txawv ntawm affinity thiab ion exchange chromatography yog qhov affinity chromatography tshwm sim vim qhov tseeb tias lub hom phiaj molecules muaj kev sib raug zoo siab rau theem nyob ruaj khov. Txawm li cas los xij, rau kev sib pauv ion chromatography, lub hom phiaj molecules muaj tus nqi rov qab rau ntawm qhov chaw nyob ruaj khov.
Cov hauv qab no qhia txog qhov sib txawv ntawm kev sib raug zoo thiab kev sib pauv ion chromatography ua ib sab ntawm kev sib piv.
Summary – Affinity vs Ion Exchange Chromatography
Nyob rau hauv cov ntsiab lus, affinity thiab ion pauv chromatography yog ob hom kua chromatographic cov tswv yim. Qhov sib txawv tseem ceeb ntawm affinity thiab ion exchange chromatography yog tias peb tuaj yeem siv affinity chromatography cais cov khoom siv hluav taws xob los yog cov khoom tsis muaj nqi hauv qhov sib tov thaum peb tuaj yeem siv ion txauv chromatography cais cov khoom sib xyaw ua ke.
