Kulekanitsa ndi kuyeretsedwa kwa mapuloteni kumagwiritsidwa ntchito kwambiri mu kafukufuku wa biochemistry ndi kugwiritsa ntchito ndipo ndi luso lofunika kwambiri.Selo lodziwika bwino la eukaryotic limatha kukhala ndi ma protein ambiri osiyanasiyana, ena ndi olemera kwambiri ndipo ena amakhala ndi makope ochepa.Kuti muphunzire zinamapuloteni, m'pofunika poyamba kuyeretsa mapuloteni kuchokera ku mapuloteni ena ndi mamolekyu omwe si a mapuloteni.

1. Njira yothetsera mcheremapuloteni:

Mchere wosalowerera ndale umakhudza kwambiri kusungunuka kwa mapuloteni.Nthawi zambiri, ndi kuchuluka kwa ndende ya mchere pansi pa mchere wochepa, kusungunuka kwa mapuloteni kumawonjezeka.Izi zimatchedwa salting;pamene ndende ya mchere ikupitiriza kuwonjezeka, Kusungunuka kwa mapuloteni kumachepa mpaka mosiyanasiyana ndikulekanitsa wina ndi mzake.Chodabwitsa ichi amatchedwa salting out.

2. Isoelectric point stacking njira:

The electrostatic repulsion pakati pa tinthu tating'onoting'ono kwambiri pamene puloteni ili static, kotero solubility ndi yaying'ono kwambiri.Ma isoelectric a mapuloteni osiyanasiyana ndi osiyana.PH ya njira yothetsera vutoli ingagwiritsidwe ntchito kuti ifike pa isoelectric point ya protein Ipangitseni kudziunjikira, koma njirayi siigwiritsidwa ntchito kawirikawiri yokha ndipo imatha kuphatikizidwa ndi njira ya salting.

3. Dialysis ndi ultrafiltration:

Dialysis imagwiritsa ntchito nembanemba yocheperako kuti ilekanitse mapuloteni amitundu yosiyanasiyana.Njira ya ultrafiltration imagwiritsa ntchito kuthamanga kwambiri kapena mphamvu ya centrifugal kupanga madzi ndi mamolekyu ena ang'onoang'ono a solute kudutsa nembanemba yotheka, pomwemapuloteniamakhala pa nembanemba.Mutha kusankha kukula kwake kosiyanasiyana kuti muchepetse mapuloteni amitundu yosiyanasiyana yama cell.

4.Gel kusefera njira:

Imatchedwanso size exclusion chromatography kapena molecular sieve chromatography, iyi ndi imodzi mwa njira zothandiza kwambiri zolekanitsira zosakaniza za mapuloteni molingana ndi kukula kwa maselo.Zida zonyamulira zomwe zimagwiritsidwa ntchito kwambiri pagawoli ndi gel osakaniza (Sephadex ged) ndi gel agarose (gel agarose).

5. Electrophoresis:

Pansi pa pH yomweyi, mapuloteni osiyanasiyana amatha kulekanitsidwa chifukwa cha kulemera kwawo kwa mamolekyulu osiyanasiyana komanso zolipiritsa zosiyanasiyana m'munda wamagetsi.Ndikoyenera kulabadira isoelectric set electrophoresis, yomwe imagwiritsa ntchito ampholyte ngati chonyamulira.Pa electrophoresis, ampholyte amapanga pH gradient pang'onopang'ono anawonjezera kuchokera elekitirodi zabwino kuti elekitirodi zoipa.Mapuloteni okhala ndi mtengo wina akasambira mmenemo, amafikirana.Malo a pH a malo amagetsi ndi olekanitsidwa, ndipo njirayi ingagwiritsidwe ntchito pofufuza ndi kukonza mapuloteni osiyanasiyana.

6.Ion kulumikizana chromatography:

Othandizira kulumikizana ndi ma ion akuphatikizapo cationic communication agents (monga carboxymethyl cellulose; CM-cellulose) ndi anionic communication agents (diethylaminoethyl cellulose).Podutsa mugawo la ion communication chromatography, puloteni yomwe ili ndi mtengo wotsutsana ndi wolumikizira ion imayikidwa pa cholumikizira cha ion, kenako ndi adsorbed.mapuloteniimasinthidwa ndikusintha pH kapena mphamvu ya ionic.

7. Affinity chromatography:

Affinity chromatography ndi njira yothandiza kwambiri yolekanitsira mapuloteni.Nthawi zambiri pamafunika sitepe imodzi yokha kuti mulekanitse puloteni inayake kuti iyeretsedwe kuchokera ku zosakaniza zosokoneza za mapuloteni ndi chiyero chapamwamba.

Njirayi imachokera pamapangidwe enieni m'malo momangirira mapuloteni ena ku molekyulu ina yotchedwa ligand (Ligand).

Mfundo yofunikira:

Mapuloteni amapezeka muzosakaniza zosakaniza mu minofu kapena maselo, ndipo mtundu uliwonse wa selo uli ndi zikwi za mapuloteni osiyanasiyana.Choncho, kusiyana pakati pa mapuloteni ndi gawo lofunika kwambiri la biochemistry, ndipo silinakhale lokha.Kapena njira zokonzekera zokonzekera zimatha kuchotsa mapuloteni amtundu uliwonse kuchokera ku mapuloteni osakanikirana, choncho njira zingapo zimagwiritsidwa ntchito nthawi zambiri.