Qhov sib txawv tseem ceeb FPLC thiab HPLC yog tias FPLC yog ib hom kua chromatography uas ntxuav cov biomolecules loj xws li cov proteins, nucleotides thiab peptides, thaum HPLC yog ib hom kua chromatography uas cais me me molecular hnyav compounds.
Ua kua chromatography yog cov txheej txheem siv los cais cov qauv rau hauv nws tus kheej cov khoom. Qhov kev sib cais no tshwm sim los ntawm kev sib cuam tshuam ntawm cov qauv tshwj xeeb nrog cov theem mobile thiab nyob ruaj khov. Cheebtsam nyob rau hauv ib tug qauv yog sib cais raws li txhua feem ntawm affinity rau lub mobile theem nyob rau hauv cov kua chromatography. Raws li cov theem ntawm lub xov tooj ntawm tes thiab cov qauv dhau los ntawm ib kem, cov khoom ntawm cov qauv pib sib cais ua pawg uas tuaj yeem kuaj pom los ntawm UV-VIS spectroscopy. Yog li, FPLC thiab HPLC yog ob hom kev ua kua chromatography.
FPLC yog dab tsi?
FPLC yog hom kua chromatography uas ntxuav cov biomolecules loj xws li cov proteins, nucleotides, thiab peptides. Qhov ceev ceev protein kua chromatography (FPLC) yog thawj zaug tsim thiab ua lag luam nyob rau hauv Sweden los ntawm Pharmacia tuam txhab nyob rau hauv 1982. Nws yog feem ntau siv los soj ntsuam los yog purify protein sib tov. Qhov no yuav siv sij hawm qhov chaw raws li lub hauv paus ntsiab lus ntawm cov qauv piv txwv affinities rau txav kua (mobile theem) thiab ib tug ntxeem tau khoom (stationary theem). Hauv FPLC, lub sijhawm txawb yog qhov tsis muaj, thiab theem nyob ruaj khov yog cov khoom siv los ntawm cov hlaws dai. Nws feem ntau muaj cov ntoo khaub lig-txuas agarose ntim rau hauv ib lub iav cylindrical lossis yas kem.
Daim duab 01: FLPC (Fast Protein Liquid Chromatography) Apparatus
Nyob rau hauv feem ntau ntawm FLPC lub tswv yim, ion pauv resin yog siv. Yog li ntawd, cov protein ntawm kev txaus siab yuav khi rau cov resin los ntawm kev sib cuam tshuam. FLPC cov txheej txheem kuj tseem siv ob lub buffers: tsis 1 (qhov khiav tsis tau) thiab tsis 2 (qhov tsis sib xws). Thaum pib, thaum khiav tsis thiab cov qauv sib tov khiav los ntawm kab, cov protein ntawm kev txaus siab hauv qhov sib tov yuav khi rau cov resin los ntawm kev sib cuam tshuam. Tab sis cov protein ntawm kev txaus siab dhau los ua kev sib cais thiab rov qab mus rau qhov kev daws teeb meem hauv elution tsis thaum lub elution buffer khiav los ntawm kem ntawm qhov kawg ntawm cov txheej txheem. Tom qab ntawd, kev daws teeb meem (eluant uas muaj protein ntau txaus siab) dhau los ntawm ob lub ntsuas, uas ntsuas ntsev concentration thiab protein concentration. Raws li txhua cov protein tau eluted, nws zoo nkaus li "ncej" thaum lub sij hawm tshawb pom thiab tuaj yeem sau tau rau kev siv ntxiv.
HPLC yog dab tsi?
HPLC yog ib hom kua chromatography uas cais cov sib txuas me me me me. Xyoo 1969, thawj HPLC tau ua lag luam los ntawm Waters Corporation, Tebchaws Asmeskas. Nyob rau hauv high-performance kua chromatography (HPLC), ib tug piv txwv sib tov (analyte) nyob rau hauv ib tug hnyav (lub sij hawm txawb) yog pumped los ntawm ib kem nrog chromatographic packing khoom (stationary theem) ntawm kev kub siab. Cov qauv sib xyaw yog nqa los ntawm kev txav cov pa roj kwj ntawm helium lossis nitrogen.
Daim duab 02: HPLC (High-Performance Liquid Chromatography)
Cov khoom hauv cov qauv sib xyaw uas muaj qhov tsawg kawg ntawm kev sib cuam tshuam nrog cov theem nyob ruaj khov lossis qhov sib cuam tshuam nrog cov theem ntawm lub xov tooj feem ntau yuav tawm ntawm kab nrawm dua. Ntawm qhov tod tes, cov khoom sib xyaw hauv cov qauv sib xyaw uas muaj qhov sib txuam nrog cov theem nyob ruaj khov lossis qhov tsawg kawg ntawm kev sib cuam tshuam nrog cov theem ntawm lub xov tooj ntawm tes yuav tawm hauv kem qeeb dua. Raws li cov txheej txheem kev sib cuam tshuam saum toj no, cov khoom ntawm cov qauv sib xyaw tuaj yeem sib cais. Tsis tas li ntawd, lub ntsuas ntsuas ntsuas ntsuas txhua yam ntawm cov qauv sib xyaw uas tawm hauv kab. HPLC yog siv los tshuaj xyuas ib puag ncig thiab lom cov qauv rau kev muaj lossis tsis muaj cov khoom sib txuas, xws li tshuaj, co toxins lossis tshuaj tua kab. Nws yog siv nyob rau hauv kev lag luam sib txawv xws li tshuaj, ib puag ncig, forensics, thiab tshuaj.
Qhov Zoo sib xws ntawm FPLC thiab HPLC yog dab tsi?
- FPLC thiab HPLC yog hom kua chromatography.
- Ob leeg siv los cais thiab txheeb xyuas cov qauv lom neeg.
- Lawv muaj cov theem ua kua mobile thiab cov theem ruaj khov.
- Ob txoj kev siv ib kem kom dhau los ntawm tus qauv.
- Cov txheej txheem no siv twj, ntsuas, li qub thiab software rau cov qauv sib cais thiab kuaj pom.
Qhov txawv ntawm FPLC thiab HPLC yog dab tsi?
FPLC yog hom kua chromatography uas yog siv los ntxuav cov biomolecules loj xws li cov proteins, nucleotides, thiab peptides. HPLC yog ib hom kua chromatography uas yog siv los cais me me molecular hnyav tebchaw. Yog li, qhov no yog qhov sib txawv tseem ceeb ntawm FPLC thiab HPLC. Ntxiv mus, FPLC siv pH thiab conductivity xyuas raws li zoo raws li ib feem ntawm cov khoom sau. Piv txwv li, HPLC tsis siv pH thiab conductivity saib xyuas thiab cov khoom siv feem ntau.
Cov duab hauv qab no qhia ntau qhov sib txawv ntawm FPLC thiab HPLC hauv daim ntawv teev npe.
Summary – FPLC vs HPLC
Chromatography yog ib lub chaw soj ntsuam cov txheej txheem siv rau kev sib cais ntawm cov khoom los ntawm kev sib tov. Nws tau muab faib ua ob peb hom xws li kem chromatography, roj chromatography, kua chromatography, ion-exchange chromatography, thiab lwm yam. FPLC thiab HPLC yog ob hom kev ua kua chromatography. FPLC yog ib hom kua chromatography uas yog siv los ntxuav cov biomolecules loj xws li cov proteins, nucleotides, thiab peptides. Ntawm qhov tod tes, HPLC yog ib hom kua chromatography uas yog siv los cais cov sib txuas me me molecular hnyav. Yog li, qhov no yog cov ntsiab lus ntawm qhov txawv ntawm FPLC thiab HPLC.
