Hoʻohana nui ʻia ka hoʻokaʻawale ʻana a me ka hoʻomaʻemaʻe ʻana o nā protein i ka noiʻi biochemistry a me ka noi ʻana a he mākaukau hana koʻikoʻi. Hiki i kekahi cell eukaryotic maʻamau ke loaʻa nā kaukani o nā protein ʻokoʻa, waiwai nui kekahi a he mau kope wale nō kekahi. I mea e aʻo ai i kekahipūmua, pono e hoʻomaʻemaʻe mua i ka protein mai nā proteins ʻē aʻe a me nā molekala non-protein.

1. Ke ano paakai opūmua:

He hopena koʻikoʻi ka paʻakai neutral i ka solubility o ka protein. ʻO ka maʻamau, me ka piʻi ʻana o ka paʻakai paʻakai ma lalo o ka haʻahaʻa o ka paʻakai paʻakai, piʻi ka solubility o ka protein. Ua kapaia keia he paakai; ke hoʻomau ka piʻi ʻana o ka paʻakai, emi ka solubility o ka protein i nā degere like ʻole a hoʻokaʻawale i kēlā me kēia. Kapa ʻia kēia ʻano he salting out.

2. Isoelectric kiko hoʻopaʻa 'ano:

ʻO ka repulsion electrostatic ma waena o nā mea liʻiliʻi ka mea liʻiliʻi loa ke kūpaʻa ka protein, no laila ʻo ka solubility ka liʻiliʻi loa. ʻOkoʻa nā helu isoelectric o nā protein like ʻole. Hiki ke ho'ohana 'ia ka pH o ka solution conditional no ka hiki 'ana i ka helu isoelectric o kahi protein E ho'onui, aka, 'a'ole hiki ke ho'ohana wale 'ia kēia ala a hiki ke ho'ohui 'ia me ke ala pa'akai.

3.Dialysis a me ka ultrafiltration:

Hoʻohana ʻo Dialysis i kahi membrane semi-permeable e hoʻokaʻawale i nā protein o nā nui molekala like ʻole. Hoʻohana ke ʻano ultrafiltration i ke kaomi kiʻekiʻe a i ʻole ka ikaika centrifugal e hana i ka wai a me nā molekole solute liʻiliʻi e hele i loko o kahi membrane semi-permeable, ʻoiai kapūmuanoho ma ka membrane. Hiki iā ʻoe ke koho i nā nui pore ʻokoʻa no ka hoʻopaʻa ʻana i nā protein o nā kaumaha molekala like ʻole.

4.Gel kānana ala:

Ua kapa ʻia hoʻi ka chromatography exclusion size a i ʻole molecular sieve chromatography, ʻo ia kekahi o nā ala pono loa no ka hoʻokaʻawale ʻana i nā huila protein e like me ka nui molekala. ʻO nā mea hoʻopili maʻamau i hoʻohana ʻia ma ke kolamu ʻo ia ka gel glucose (Sephadex ged) a me ka gel agarose (gel agarose).

5.Electrophoresis:

Ma lalo o ke kūlana pH like, hiki ke hoʻokaʻawale ʻia nā protein like ʻole ma muli o kā lākou mau paona molekala like ʻole a me nā uku like ʻole i ke kahua uila. Pono e hoʻolohe i ka electrophoresis hoʻonohonoho isoelectric, e hoʻohana ana i kahi ampholyte ma ke ʻano he mea lawe. I ka wā electrophoresis, hoʻokumu ka ampholyte i kahi gradient pH i hoʻohui mālie ʻia mai ka electrode maikaʻi i ka electrode maikaʻi ʻole. I ka wā e ʻau ai ka protein me kahi uku i loko o laila, e hōʻea kekahi i kekahi. Hoʻopau ʻia ke kūlana pH o ke kiko uila, a hiki ke hoʻohana ʻia kēia ʻano no ka nānā ʻana a hoʻomākaukau i nā protein like ʻole.

6. Ion kamaʻilio chromatography:

Hoʻokomo ʻia nā ʻelele kamaʻilio ion i nā mea kamaʻilio cationic (e like me ka carboxymethyl cellulose; CM-cellulose) a me nā mea kamaʻilio anionic (diethylaminoethyl cellulose). I ka hele ʻana i ke kolamu chromatography kamaʻilio ion, ua hoʻopili ʻia ka protein me ka uku ʻē aʻe i ka ʻelele kamaʻilio ion ma luna o ka mea kamaʻilio ion, a laila hoʻopili ʻia ka adsorbed.pūmuaeluted ma ka hoololi ana i ka pH a ionic ikaika.

7. Affinity chromatography:

ʻO ka chromatography affinity kahi ala kūpono loa no ka hoʻokaʻawale ʻana i nā protein. Hoʻokahi wale nō ʻanuʻu e hoʻokaʻawale i kahi protein e hoʻomaʻemaʻe ʻia mai kahi hui pūmua me ka maʻemaʻe kiʻekiʻe.

Hoʻokumu ʻia kēia ʻano hana ma kahi kikoʻī ma mua o ka paʻa covalent o kekahi mau protein i kekahi mole i kapa ʻia he ligand (Ligand).

ʻO ke kumu kumu:

Aia nā protein i loko o kahi hui ʻino i loko o nā ʻiʻo a i ʻole nā ​​​​pūnaewele, a ʻo kēlā me kēia ʻano o ke kelepona he mau kaukani mau protein ʻokoʻa. No laila, ʻo ka ʻokoʻa ma waena o nā protein he ʻāpana koʻikoʻi o ka biochemistry, a ʻaʻole ʻo ia wale nō. A i ʻole kahi hoʻonohonoho o nā ʻano hana i mākaukau hiki ke wehe i kekahi ʻano protein mai kahi protein i hui pū ʻia, no laila e hoʻohana pinepine ʻia nā ʻano hana like ʻole.