Cov txheej txheem ntawmprotein purification:

Cov txheej txheem ntawm cov protein purification, sib cais thiab purification ntawm cov protein, cov protein yog tso tawm los ntawm lub qub hlwb los yog cov ntaub so ntswg nyob rau hauv ib tug yaj lub xeev thiab tseem nyob rau hauv lub qub natural lub xeev yam tsis muaj kev ua si lom. Vim li no, cov khoom siv yuav tsum tshem tawm cov tsiaj txhu thiab cov ntaub so ntswg adipose, cov khoom siv noob yuav tsum txawm tev tawm cov kab mob ntawm cov noob los ntawm cov khoom xws li tannins, thiab cov noob roj yuav tsum nyiam siv cov kuab tshuaj uas tsis muaj dej kub npau npau.

1. Cov txheej txheem ntawm cov protein purification, qhov loj me thiab kev sib cais sib txawv: kev lim ntshav thiab ultrafiltration siv cov proteins kom dhau los ntawm qhov xwm txheej ntawm cov molecules uas tsis tuaj yeem dhau los ntawm cov txheej txheem semi-permeable; density gradient centrifugation Qhov zoo thiab qhov ntom ntawm cov protein nyob hauv nruab nrab yuav muaj kev cuam tshuam ntau dua rau cov khoom. Kev daws teeb meem nrawm dua; gel filtration yog ib hom kab chromatography.

2. Kev sib cais ntawm solubility sib txawv ntawm kev siv: electro- nag lossis daus vim hais tias tus nqi ntawm cov protein molecules yog zero isoelectric point, txo cov electrostatic repulsion ntawm molecules, yooj yim rau aggregate, thiab yam tsawg kawg nkaus solubility; ntsev siv nrog ntsev tov Cov concentration nce los yog txo cov solubility ntawm cov protein.

3. Cov nqi sib cais thiab tshuaj xyuas los ntawm ntau txoj kev, cov ntsiab lus tseem ceeb suav nrog electrophoresis thiab ion exchange chromatography.

4. Cov txheej txheem ntawmprotein purificationKev xaiv adsorption thiab sib cais ntawm cov proteins siv lub zog ntawm particle adsorption.

5. Affinity chromatography yog raws li kev sib cais ntawm ligands. Cov yam ntxwv lom neeg no tso cai rau cov protein molecules los khi tshwj xeeb es tsis yog covalently rau lwm cov molecule hu ua ligand.

6. Txoj kev ua kom huv ntawm cov protein: Cov txheej txheem qis-kub organic solvent nag lossis daus: Siv cov kuab tshuaj organic uas muaj miscible nrog dej, xws li methanol, ethanol, kom txo cov solubility thiab nag lossis daus ntawm cov protein ntau. Txoj kev no yog siab tshaj qhov kev daws teeb meem salting tawm, tab sis cov protein ntau denatures Thaum yooj yim, nws yuav tsum tau ua ntawm qhov kub thiab txias.

Yuav ua li cas ntxuavprotein purification:

Yuav ua li cas purify protein, protein yog ib tug loj molecule, thiab ntau yam molecular ntau thiab tsawg ntawm albumin, muaj ib co yooj yim txoj kev los cais cov protein thiab cov khoom me me, thiab tseem cais cov protein sib xyaw. Cov txheej txheem tseem ceeb rau kev sib cais cov protein ntawm qhov sib txawv ntawm qhov ntau thiab tsawg yog kev lim ntshav, ultrafiltration, gel filtration, thiab kev sib cais centrifugal. Dialysis thiab ultrafiltration feem ntau yog siv rau hauv txoj kev sib cais protein. Cov tshuaj lim dej sib cais yuav tsum tau muab tso rau hauv ib lub hnab lim dej ua los ntawm cov khoom siv semipermeable, thiab tom qab ntawd muab tso rau hauv cov tshuaj lim dej rau kev sib cais. Ultrafiltration yog cov txheej txheem ntawm kev hla dej thiab lwm cov molecules me me uas siv lub zog centrifugal los yog siab nyob rau hauv ib tug semi-permeable membrane thiab trapping proteins nyob rau hauv lub semi-permeable membrane. Ob txoj kev no tuaj yeem sib cais los ntawm cov protein-based macromolecules thiab inorganic ntsev-raws li me me molecules. Lawv feem ntau salted tawm, siv nrog rau cov ntsev dissolution txoj kev, los yog yuav ua tswvcuab coj tawm ob txoj kev tom qab salting tawm siv inorganic ntsev tov.

1. Yuav ua li cas purify protein los ntawm salting-tawm txoj kev: lub hauv paus ntawm salting-tawm txoj kev yog solubility ntawm cov protein nyob rau hauv dilute ntsev tov yuav nce nrog nce ntsev concentration, tab sis thaum lub ntsev concentration nce mus rau ib tug tej yam nqi, lub kev ua dej num yuav nce ntxiv. Txo, uas nyob rau hauv lem tuaj yeem ua rau qee cov proteins los tsim cov nqi los ntawm cov qauv molecular kom maj mam neutralized, thiab cov hydration membrane yog maj mam rhuav tshem, uas tuaj yeem ua rau muaj feem xyuam rau cov proteins thiab cov molecules los tsim agglomeration thiab precipitate los ntawm cov kev daws teeb meem sib txawv.

2. Organic solvent precipitation method: Muaj ob qho laj thawj los txo qhov solubility ntawm protein nyob rau hauv cov kuab tshuaj organic: yuav ua li cas purify protein raws li ntsev tov nrog lub cev qhuav dej; Thib ob, qhov dielectric tas li ntawm cov kuab tshuaj organic dua li dej tuaj yeem txo qis hauv cov kuab tshuaj polar.

3. Protein precipitating agent: protein raws li ib tug precipitating tus neeg saib xyuas tsuas yog ua rau ib hom los yog ib tug ntawm cov protein los nag, uas yog alkaline, protein, lectin, thiab hnyav hlau.

4. Polyethylene glycol nag lossis daus: Yuav ua li cas ntxuav cov dej-soluble non-ionic polymers rauprotein purification, xws li polyethylene glycol thiab sodium dextran sulfate rau precipitate proteins.