Gutandukanya no kweza poroteyine bikoreshwa cyane mubushakashatsi bwibinyabuzima no kubukoresha kandi ni ubuhanga bukomeye bwo gukora. Ingirabuzimafatizo ya eukaryotique irashobora kuba irimo poroteyine ibihumbi n'ibihumbi, zimwe zirakize cyane kandi zirimo kopi nkeya. Kugirango wige ikintu runakaporoteyine, ni ngombwa kubanza kweza poroteyine mu zindi poroteyine na molekile zitari poroteyine.

1. Uburyo bwo gushunguraporoteyine:

Umunyu udafite aho ubogamiye ugira ingaruka zikomeye ku gukomera kwa poroteyine. Mubisanzwe, hamwe no kwiyongera kwumunyu mwinshi mukunyunyu muke, gukomera kwa poroteyine biriyongera. Ibi byitwa umunyu; iyo umunyu ukomeje kwiyongera, Ubushobozi bwa poroteyine bugabanuka kuburyo butandukanye kandi bugatandukanya umwe umwe. Iki kintu cyitwa umunyu.

2. Uburyo bwo gutondekanya ingingo ya Isoelectric:

Kwanga electrostatike hagati yuduce ni duto iyo poroteyine ihagaze, bityo solubile nayo ikaba nto. Ingingo ya isoelectric ya proteine ​​zitandukanye ziratandukanye. PH yumuti wibisubizo urashobora gukoreshwa kugirango ugere kuri isoelectric point ya proteine ​​Bitume yegeranya, ariko ubu buryo ntibukoreshwa gake kandi burashobora guhuzwa nuburyo bwo gusohora.

3.Dialysis na ultrafiltration:

Dialysis ikoresha igice cya kabiri cyoroshye kugirango itandukane poroteyine zingana na molekile zitandukanye. Uburyo bwa ultrafiltration bukoresha umuvuduko mwinshi cyangwa imbaraga za centrifugal kugirango amazi nizindi molekile ntoya ya solute inyure muri kimwe cya kabiri cyinjira, mugiheporoteyinekuguma kuri membrane. Urashobora guhitamo ubunini butandukanye bwa pore kugirango uhagarike poroteyine zipima uburemere butandukanye.

4.Uburyo bwo kuyungurura:

Byitwa kandi ubunini bwo gutandukanya chromatografiya cyangwa molekile ya sikeri ya chromatografiya, ubu ni bumwe muburyo bwingenzi bwo gutandukanya imvange ya poroteyine ukurikije ubunini bwa molekile. Ibikoresho bikoreshwa cyane mu gupakira mu nkingi ni glucose gel (Sephadex ged) na gel agarose (gel agarose).

5.Electrophoresis:

Muburyo bumwe bwa pH, poroteyine zitandukanye zirashobora gutandukana bitewe nuburemere bwa molekile zitandukanye hamwe nuburyo butandukanye mumashanyarazi. Birakwiye ko twita kuri isoelectric set electrophoresis, ikoresha ampholyte nkitwara. Mugihe cya electrophorei, ampholyte ikora pH gradient yongeweho buhoro buhoro kuva kuri electrode nziza kuri electrode mbi. Iyo poroteyine ifite charge runaka yoga muri yo, izageraho. Umwanya wa pH wumurongo wamashanyarazi urahagarara, kandi ubu buryo burashobora gukoreshwa mugusesengura no gutegura proteine ​​zitandukanye.

6.Mu itumanaho rya chromatografiya:

Ibikoresho byitumanaho ion birimo ibikoresho byitumanaho (nka carboxymethyl selulose; CM-selile) hamwe nubutumwa bwitumanaho rya anionic (diethylaminoethyl selulose). Iyo unyuze muri ion itumanaho rya chromatografiya, poroteyine hamwe nuburyo bunyuranye kubakozi ba ion itumanaho byamamazwa kuri ion itumanaho, hanyuma bikamenyekana.poroteyineisubirwamo muguhindura imbaraga za pH cyangwa ionic.

7.Ubusobanuro bwa chromatografiya:

Affinity chromatography nuburyo bwingirakamaro cyane bwo gutandukanya poroteyine. Bikunze gusaba intambwe imwe gusa yo gutandukanya poroteyine runaka kugirango isukure ivanze na poroteyine ivanze nubuziranenge bwinshi.

Ubu buryo bushingiye ku buryo bwihariye aho guhuza poroteyine zimwe na zimwe kuri molekile yitwa ligand (Ligand).

Ihame shingiro:

poroteyine zibaho mu ruvangitirane ruvanze mu ngingo cyangwa selile, kandi buri bwoko bwa selile burimo poroteyine ibihumbi. Kubwibyo, gutandukanya poroteyine nigice cyingenzi cyibinyabuzima, kandi ntabwo byabaye wenyine. Cyangwa uburyo bwuburyo bwateguwe burashobora gukuraho ubwoko bwose bwa poroteyine muri poroteyine ivanze, bityo uburyo bwinshi bukoreshwa muguhuza.