A na-eji nkewa na ịdị ọcha nke protein n'ọtụtụ ebe na nyocha na ngwa biochemistry na ọ bụ nka dị mkpa nke ịrụ ọrụ. Otu mkpụrụ ndụ eukaryotic a na-ahụkarị nwere ike ịnwe puku kwuru puku protein dị iche iche, ụfọdụ bara ụba nke ukwuu na ụfọdụ nwere naanị ole na ole. Iji mụọ ụfọdụprotein, ọ dị mkpa ibu ụzọ mee ka protein dị ọcha site na protein ndị ọzọ na ụmụ irighiri ihe na-abụghị protein.

1. Nnu-esi ụzọ nkeprotein:

Nnu na-anọpụ iche nwere mmetụta dị ukwuu na solubility nke protein. N'ozuzu, na mmụba nke ntinye nnu n'okpuru ntanye nnu dị ala, ihe solubility nke protein na-abawanye. Nke a na-akpọ nnu; mgbe ntinye nnu na-aga n'ihu na-abawanye, Solubility nke protein na-ebelata ruo ogo dị iche iche ma kewapụ otu mgbe. A na-akpọ ihe a na-eme nnu nnu.

2. Usoro nchịkọta isi ihe nke isoelectric:

The electrostatic repulsion n'etiti ụmụ irighiri ihe bụ nke kasị nta mgbe protein bụ static, otú solubility bụkwa nke kasị nta. Isi ihe isoelectric nke protein dị iche iche dị iche iche. Enwere ike iji pH nke ihe ngwọta na-edozi ahụ iji ruo ebe isoelectric nke protein Mee ka ọ gbakọta, ma usoro a adịghị adịkarị eji naanị ya ma nwee ike jikọta ya na usoro nnu.

3. Dialysis na ultrafiltration:

Dialysis na-eji akpụkpọ ahụ nke nwere ike ime ka ọ kewapụ protein dị iche iche nha molekụla. Usoro ultrafiltration na-eji nrụgide dị elu ma ọ bụ ike centrifugal mee ka mmiri na obere ụmụ irighiri ihe ndị ọzọ na-agafe n'ime akpụkpọ ahụ nke na-agafe agafe, ebeproteinna-anọgide na akpụkpọ ahụ. Ị nwere ike ịhọrọ nha pore dị iche iche iji gbochie protein dị arọ dị iche iche.

4.Gel filtration usoro:

A na-akpọkwa chromatography mwepu nha ma ọ bụ chromatography sieve molecular, nke a bụ otu n'ime ụzọ kachasị baa uru maka ikewa ngwakọta protein dị ka nha molekul siri dị. Ihe ngwugwu a na-ejikarị na kọlụm bụ glucose gel (Sephadex ged) na agarose gel (agarose gel).

5. Electrophoresis:

N'okpuru otu ọnọdụ pH, enwere ike kewapụ protein dị iche iche n'ihi ịdị arọ ha dị iche iche na ụgwọ dị iche iche na mpaghara eletrik. Ọ bara uru ịṅa ntị na isoelectric set electrophoresis, nke na-eji ampholyte dị ka onye na-ebu. N'oge electrophoresis, ampholyte na-etolite pH gradient nke nta nke nta gbakwunyere site na electrode dị mma na electrode na-adịghị mma. Mgbe protein nwere ụgwọ ụfọdụ na-egwu mmiri na ya, ọ ga-eru ibe ya. Ọnọdụ pH nke ebe eletrik na-akwụsị, a pụkwara iji usoro a nyochaa na ịkwadebe protein dị iche iche.

6.Ion nkwurịta okwu chromatography:

Ndị ọrụ nkwurịta okwu ion gụnyere ndị ọrụ nkwurịta okwu cationic (dị ka carboxymethyl cellulose; CM-cellulose) na ndị ọrụ nkwurịta okwu anionic (diethylaminoethyl cellulose). Mgbe ị na-agafe na kọlụm nkwurịta okwu ion chromatography, protein nke nwere ụgwọ dị iche na onye na-ahụ maka nkwurịta okwu ion na-etinye ya na onye na-ahụ maka nkwurịta okwu ion, wee gbasaa ya.proteinA na-ewepụ ya site na ịgbanwe pH ma ọ bụ ike ionic.

7. Affinity chromatography:

Affinity chromatography bụ ụzọ bara uru nke ukwuu maka ikewa protein. Ọ na-achọkarị naanị otu nzọụkwụ iji kewaa ụfọdụ protein ka a sachaa ya na ngwakọta protein na-adịghị mma nke nwere ịdị ọcha dị elu.

Usoro a gbadoro ụkwụ na nke a kapịrị ọnụ kama ijikọ protein ụfọdụ na molekul ọzọ a na-akpọ ligand (Ligand).

Ụkpụrụ bụ isi:

protein dị na ngwakọta na-adịghị mma na anụ ahụ ma ọ bụ sel, na ụdị cell nke ọ bụla nwere ọtụtụ puku protein dị iche iche. Ya mere, ọdịiche dị n'etiti protein bụ akụkụ dị mkpa nke biochemistry, ọ bụghị naanị ya. Ma ọ bụ usoro a na-emezi nke ọma nwere ike iwepu ụdị protein ọ bụla na protein agwakọtara adịghị mma, ya mere a na-ejikarị ọtụtụ ụzọ eme ihe.