Ukuhlukaniswa nokuhlanzwa kwamaprotheni kusetshenziswa kakhulu ocwaningweni nasekusetshenzisweni kwe-biochemistry futhi kuyikhono elibalulekile lokusebenza. Iseli elijwayelekile le-eukaryotic lingaqukatha izinkulungwane zamaphrotheni ahlukene, amanye acebe kakhulu kanti amanye aqukethe amakhophi ambalwa kuphela. Ukuze ufunde okuthileamaprotheni, kuyadingeka ukuhlanza amaprotheni kuqala kwamanye amaprotheni nama-molecule angewona amaprotheni.

1. Indlela yokukhipha usawotiamaprotheni:

Usawoti ongathathi hlangothi unomthelela omkhulu ekuncibilikeni kwamaprotheni. Ngokuvamile, ngokwanda kokuhlushwa kukasawoti ngaphansi kokuhlushwa kukasawoti omncane, ukuncibilika kwamaprotheni kuyakhula. Lokhu kubizwa nge-salting; lapho ukuhlushwa kukasawoti kuqhubeka nokwanda, I-solubility yamaprotheni iyancipha ibe ngamadigri ahlukahlukene futhi ihlukanise enye ngemva kwenye. Lesi simo sibizwa ngokuthi i-salting out.

2. Indlela yokunqwabelanisa iphoyinti le-isoelectric:

I-electrostatic repulsion phakathi kwezinhlayiya incane kakhulu uma iphrotheni imile, ngakho-ke ukunyibilika nakho kukuncane kakhulu. Amaphuzu e-isoelectric amaprotheni ahlukahlukene ahlukile. I-pH yesisombululo se-conditioning ingasetshenziswa ukufinyelela iphuzu le-isoelectric leprotheni Yenze inqwabelene, kodwa le ndlela ayivamile ukusetshenziswa iyodwa futhi ingahlanganiswa nendlela yokukhipha usawoti.

3.I-Dialysis kanye ne-ultrafiltration:

I-Dialysis isebenzisa ulwelwesi olukwazi ukungena kalula ukuze luhlukanise amaprotheni amasayizi amangqamuzana ahlukene. Indlela ye-ultrafiltration isebenzisa ingcindezi ephezulu noma amandla aphakathi nendawo ukwenza amanzi namanye ama-athomu amancane ancibilike adlule kulwelwesi olukwazi ukungena kancane, kuyilaphoamaprotheniihlala kulwelwesi. Ungakhetha osayizi abahlukene bezimbotshana ukuze ubambe amaprotheni ezisindo zamangqamuzana ahlukene.

4.Indlela yokuhlunga yejeli:

Ebizwa nangokuthi i-chromatography yokukhishwa kosayizi noma i-molecular sieve chromatography, lena enye yezindlela eziwusizo kakhulu zokuhlukanisa izingxube zamaprotheni ngokuya ngosayizi wamangqamuzana. Izinto zokupakisha ezisetshenziswa kakhulu kukholamu ijeli yeglucose (Sephadex ged) kanye nejeli ye-agarose (ijeli ye-agarose).

5.I-Electrophoresis:

Ngaphansi kwesimo esifanayo se-pH, amaprotheni ahlukahlukene angahlukaniswa ngenxa yesisindo sawo samangqamuzana ahlukene kanye namashaji ahlukene emkhakheni kagesi. Kuyafaneleka ukunaka isethi ye-isoelectric electrophoresis, esebenzisa i-ampholyte njengesithwali. Ngesikhathi se-electrophoresis, i-ampholyte yakha i-pH gradient kancane kancane eyengezwe isuka ku-electrode eyakhayo iye ku-electrode engemihle. Lapho iphrotheni eneshaja ethile ibhukuda kuyo, izofinyelela komunye nomunye. Indawo ye-pH yendawo kagesi ayiqhubeki, futhi le ndlela ingasetshenziswa ukuhlaziya nokulungisa amaprotheni ahlukahlukene.

I-chromatography yokuxhumana ye-Ion:

ama-ejenti okuxhumana nge-ion ahlanganisa ama-agent okuxhumana e-cationic (afana ne-carboxymethyl cellulose; i-CM-cellulose) kanye nama-anionic communication agents (i-diethylaminoethyl cellulose). Lapho udlula kukholamu yechromatography yokuxhumana kwe-ion, iphrotheni eneshaji eliphambene ne-ejenti yokuxhumana ye-ion ikhangiswe kumenzeli wokuxhumana we-ion, bese kuba i-adsorbed.amaprotheniikhishwa ngokushintsha i-pH noma amandla e-ionic.

7.Affinity chromatography:

I-Affinity chromatography iyindlela ewusizo kakhulu yokuhlukanisa amaprotheni. Ngokuvamile kudinga isinyathelo esisodwa kuphela ukuhlukanisa iphrotheni ethile ukuze ihlanzwe engxubeni yamaprotheni engcolile enobumsulwa obuphezulu.

Le ndlela isekelwe ekubopheni okuqondile kunokuvumelana kwamaprotheni athile kwenye i-molecule ebizwa ngokuthi i-ligand (Ligand).

Umgomo oyisisekelo:

amaprotheni akhona engxubeni engcolile ezicutshini noma kumaseli, futhi uhlobo ngalunye lweseli luqukethe izinkulungwane zamaphrotheni ahlukene. Ngakho-ke, umehluko phakathi kwamaprotheni uyingxenye ebalulekile ye-biochemistry, futhi ayizange ibe yodwa. Noma isethi yezindlela esezilungile zingasusa noma yiluphi uhlobo lwamaprotheni ephrotheni exubene engcolile, ngakho-ke izindlela ezimbalwa zivame ukusetshenziswa ngokuhlanganiswa.