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 ephambene 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.