Kupatsanurwa uye kucheneswa kwemapuroteni kunoshandiswa zvakanyanya mukutsvagisa biochemistry uye kushandisa uye yakakosha hunyanzvi hwekuita. Sero rakajairwa eukaryotic rinogona kuve nezviuru zvemapuroteni akasiyana, mamwe akapfuma kwazvo uye mamwe ane mashoma makopi. Kuti udzidze chimwe chinhuprotein, zvakakosha kutanga kuchenesa mapuroteni kubva kune mamwe mapuroteni uye asiri mapuroteni molecules.

1. Salting-out nzira yeprotein:

Munyu usina kwawakarerekera une simba guru pakunyungudika kweprotein. Kazhinji, nekuwedzera kwekuiswa kwemunyu pasi pemunyu wakaderera, kunyungudika kweprotein kunowedzera. Izvi zvinonzi salting; apo munyu wemunyu unoramba uchiwedzera, Kunyungudika kweprotein kunoderera kusvika kune madhigirii akasiyana uye kunoparadzanisa imwe mushure meimwe. Chiitiko ichi chinonzi salting out.

2. Isoelectric point stacking nzira:

Iyo electrostatic repulsion pakati pezvimedu ndiyo idiki kana protein yakamira, saka kunyungudika kudiki zvakare. Iwo isoelectric mapoinzi emapuroteni akasiyana akasiyana. Iyo pH yemushonga wekugadzirisa inogona kushandiswa kusvika kune isoelectric point yeprotein Iite kuti iunganidze, asi iyi nzira haiwanzo shandiswa yoga uye inogona kusanganiswa neiyo salting-out nzira.

3. Dialysis uye ultrafiltration:

Dialysis inoshandisa semi-permeable membrane kuparadzanisa mapuroteni emhando dzakasiyana dzemamorekuru. Iyo ultrafiltration nzira inoshandisa yakanyanya kudzvanywa kana simba recentrifugal kuita kuti mvura uye mamwe madiki solute mamorekuru apfuure nepakati semi-permeable membrane, nepoproteininoramba iri pa membrane. Iwe unogona kusarudza hukuru hwakasiyana hwepore kubata mapuroteni ehuremu hwemamorekuru akasiyana.

4.Gel kusefa nzira:

Iyo inonziwo saizi yekusarudzika chromatography kana molecular sieve chromatography, iyi ndiyo imwe yenzira dzinobatsira dzekuparadzanisa maprotein musanganiswa zvichienderana nekukura kwemamorekuru. Iyo inonyanya kushandiswa kurongedza zvinhu mukoramu ndeye glucose gel (Sephadex ged) uye agarose gel (agarose gel).

5.Electrophoresis:

Pasi pemamiriro akafanana epH, mapuroteni akasiyana anogona kupatsanurwa nekuda kwehuremu hwawo hwakasiyana hwemamorekuru uye machaji akasiyana mumunda wemagetsi. Zvakakosha kubhadhara kune isoelectric set electrophoresis, iyo inoshandisa ampholyte semutakuri. Munguva ye electrophoresis, ampholyte inoumba pH gradient zvishoma nezvishoma inowedzerwa kubva kune yakanaka electrode kusvika kune yakaipa electrode. Apo puroteni ine imwe charge inoshambira mairi, inosvika kune imwe neimwe. Iyo pH chinzvimbo chegetsi poindi inoregedza, uye iyi nzira inogona kushandiswa kuongorora nekugadzirira akasiyana mapuroteni.

6.Ion kutaurirana chromatography:

Ion kutaurirana vamiririri vanosanganisira cationic kutaurirana vamiririri (senge carboxymethyl cellulose; CM-cellulose) uye anionic kutaurirana vamiririri (diethylaminoethyl cellulose). Kana uchipfuura neiyo ion yekutaurirana chromatography column, puroteni ine yakatarisana nechaji kune iyoion yekutaurirana agent inoiswa paion communication agent, uyezve adsorbed.proteininokwenenzverwa nekushandura pH kana ionic simba.

7. Affinity chromatography:

Affinity chromatography inzira inobatsira kwazvo yekuparadzanisa mapuroteni. Zvinowanzoda nhanho imwe chete kuparadzanisa imwe puroteni kuti icheneswe kubva kune yakasvibiswa musanganiswa weprotein nekuchena kwepamusoro.

Iyi nzira yakavakirwa pane chaiyo kwete covalent kusunga kwemamwe mapuroteni kune imwe morekuru inonzi ligand (Ligand).

The basic principle:

mapuroteni aripo mumusanganiswa wakashata mumatishu kana maseru, uye mhando yese yesero ine zviuru zvemapuroteni akasiyana. Naizvozvo, musiyano pakati pemapuroteni chikamu chakakosha chebiochemistry, uye haina kunge iri yega. Kana kuti seti yenzira dzakagadzirirwa-yakagadzirirwa dzinogona kubvisa chero rudzi rweprotein kubva kune yakashata yakasanganiswa protein, saka nzira dzinoverengeka dzinowanzoshandiswa pamwe chete.