Iindlela zeukuhlanjululwa kweeprotheyini:

Indlela yokuhlanjululwa kweprotheyini, ukwahlukana kunye nokuhlanjululwa kweprotheni, iprotheni ikhutshwa kwiiseli zangaphambili okanye izicubu kwimeko echithwe kwaye ihlala kwindawo yokuqala yendalo ngaphandle kokulahlekelwa kwemisebenzi yezinto eziphilayo. Ngenxa yesi sizathu, izinto eziphathekayo kufuneka zisuse isilwanyana kunye nezicubu ze-adipose, izinto zembewu kufuneka zikhuphe ukungcoliseka kwengubo yembewu kwizinto ezifana ne-tannins, kunye nembewu yeoli kufuneka isebenzise i-solvent ephantsi yokubilisa.

1. Indlela yokuhlanjululwa kweprotheyini, ubungakanani kunye neendlela ezahlukeneyo zokuhlalutya ukuhlukana: i-dialysis kunye ne-ultrafiltration isebenzisa ezi proteni ukuba zidlule kwindalo yeemolekyuli ezingenakudlula kwi-membrane ye-semi-permeable; Ubungakanani beprotheyini ephakathi iya kuba nempembelelo enkulu kumasuntswana. Isantya sokumisa ngokukhawuleza; ukuhluzwa kwejeli luhlobo lwekholamu yechromatography.

2. Ukwahlula umahluko we-solubility ekusetyenzisweni: i-electro-precipitation ngenxa yokuba i-net intlawulo yeemolekyuli zeprotheni ikwindawo ye-isoelectric ye-zero, ukunciphisa ukunyanzeliswa kwe-electrostatic phakathi kweeamolekyu, kulula ukuhlanganisa, kunye ne-solubility encinci; isisombululo setyuwa esisetyenziswe kunye nesisombululo setyuwa Ukugxila kwandisa okanye kunciphisa ukunyibilika kweprotheni.

3. Umrhumo uhlukaniswe kwaye uhlalutywe ngeendlela ezahlukeneyo, iziqulatho eziphambili ziquka i-electrophoresis kunye ne-ion exchange chromatography.

4. Iindlela zeukuhlanjululwa kweeprotheyiniI-adsorption ekhethiweyo kunye nokwahlulwa kweeprotheni zisebenzisa amandla e-particle adsorption.

5. I-chromatography yobudlelwane isekelwe ekuhlukaneni kwe-ligands. Olu phawu lwebhayoloji luvumela iimolekyuli zeprotheyini ukuba zibophe ngokuthe ngqo kunokuba zidibanise enye imolekyuli ebizwa ngokuba yi-ligand.

6. Indlela yokuhlanjululwa kweprotheyini: Ubushushu obuphantsi be-organic solvent precipitation method: Sebenzisa i-solvents eziphilayo ezixutywe ngamanzi, ezifana ne-methanol, i-ethanol, ukunciphisa ukunyibilika kunye nemvula yeeprotheni ezininzi. Le ndlela iphezulu kunesisombululo se-salting ngaphandle, kodwa iprotheni i-denatures ngakumbi Xa kulula, kufuneka yenziwe kwiqondo lokushisa eliphantsi.

Indlela yokucocaukuhlanjululwa kweeprotheyini:

Indlela yokucoca iprotheni, iprotheni yimolekyuli enkulu, kunye nobukhulu obuhlukeneyo beemolekyuli ze-albumin, kukho iindlela ezilula zokwahlula iiprotheni kunye nezixhobo ezincinci ze-molecule, kunye nomxube weprotheni owahlulayo. Iindlela eziphambili zokwahlula iiprotheyini zobungakanani beemolekyuli ezahlukeneyo yi-dialysis, i-ultrafiltration, i-gel filtration, kunye nokwahlulwa kwe-centrifugal. I-Dialysis kunye ne-ultrafiltration isetyenziswa ngokuqhelekileyo kwiindlela zokuhlukana kweeprotheni. Umxube we-dialysis oza kuhlukaniswa ufakwe kwisikhwama se-dialysis esenziwe kwi-membrane e-semipermeable, emva koko ifakwe kwisisombululo se-dialysis sokwahlula. I-Ultrafiltration yinkqubo yokudlula kwamanzi kunye nezinye iamolekyu ezincinci zisebenzisa amandla e-centrifugal okanye uxinzelelo kwi-membrane ye-semi-permeable kunye nokubambisa iiprotheni kwi-membrane ye-semi-permeable. Ezi ndlela zimbini zinokwahlulwa kwii-macromolecules ezisekelwe kwiprotheyini kunye neemolekyuli ezincinci ezisekelwe kwityuwa. Zihlala zigalelwe ityuwa ngaphandle, zisetyenziswe kunye nendlela yokuyinyibilikisa ityuwa, okanye zinokuziswa ukuze zikhuphe ezi ndlela zimbini emva kokufakwa kwetyuwa kusetyenziswa izisombululo zetyuwa ezingaphiliyo.

1. Indlela yokuhlambulula iprotheni ngendlela ye-salting-out: Isiseko sendlela ye-salting-out kukuba ukunyibilika kweprotheni kwisisombululo setyuwa edibeneyo kuya kwanda ngokunyuka kwetyuwa, kodwa xa ityuwa inyuka ukuya kwixabiso elithile, umsebenzi wamanzi uya kwandiswa. Ukuncipha, okunokuthi kubangele ukuba ezinye iiproteni zenze iintlawulo ngomphezulu wemolekyuli ukuba zingathathi hlangothi ngokuthe ngcembe, kwaye i-membrane ye-hydration itshatyalaliswe ngokuthe ngcembe, enokuthi ekugqibeleni ibangele iiproteni ezinxulumene neemolekyuli ukuba ziphuhlise i-agglomeration kunye nemvula evela kwizisombululo ezahlukeneyo.

2. Indlela yemvula ye-solvent ye-Organic: Kukho izizathu ezibini zokunciphisa i-solubility yeprotheyini kwi-solvents yezinto eziphilayo: indlela yokucoca iprotheni njengesisombululo setyuwa kunye nokuchithwa kwamanzi; okwesibini, i-dielectric constant of organic solvents kunamanzi inokuncitshiswa Kwi-solvents ye-polar.

3. I-protein precipitating agent: iprotheni njenge-precipitating agent isebenza kuphela kuhlobo olulodwa okanye enye ye-protein precipitation, eziqhelekileyo ziyi-alkaline, iprotheni, i-lectin, kunye neentsimbi ezinzima.

4. Imvula yePolyethylene glycol: Uzicoca njani iipolima ezinyibilikayo ezingena-ionic emanziniukuhlanjululwa kweeprotheyini, ezifana ne-polyethylene glycol kunye ne-sodium dextran sulfate ukukhupha iiprotheni.