Ukusetshenziswa kwe-ammonium sulfate protein purification

Ukusetshenziswa kwe-ammonium sulfate ekuhlanzeni amaprotheni

 

Amaprotheni ayizinto ezibalulekile zemisebenzi yempilo, futhi ukwehluka kwawo ngesakhiwo kanye nokusebenza kunquma ukubaluleka kwawo ku-biomedicine, ucwaningo lwebhayoloji yamangqamuzana, kanye nokukhiqizwa kwezimboni. Ukuhlukaniswa nokuhlanzwa kwamaprotheni kuyizinyathelo ezibalulekile emkhakheni we-biotechnology, futhi i-ammonium sulfate, njenge-precipitant yamaprotheni esetshenziswa kakhulu, idlala indima ebalulekile enqubweni yokuhlanzwa kwamaprotheni.

 

1. Ukubaluleka kokuhlanza amaprotheni

 

Ukuhlanzwa kwamaprotheni kuyisidingo sokuqala sokuqinisekisa ukunemba kocwaningo lwamaprotheni nokusebenzisa. Amaprotheni ahlanzekile angasetshenziselwa:

 

Ukuhlaziywa kwesakhiwo, njenge-X-ray crystallography, i-nuclear magnetic resonance (NMR), njll.

 

Ucwaningo olusebenzayo, olufana ne-enzyme kinetics, izici ezibophayo, njll.

 

Ukuthuthukiswa kwezidakamizwa: njengethagethi yezidakamizwa noma umuthi ngokwawo.

 

Izicelo zezimboni: njengama-catalysts, izithasiselo zokudla, njll.

 

2. Indima ye-ammonium sulfate ekuhlanzeni amaprotheni

 

I-Ammonium sulfate (NH4) 2SO4) ingusawoti wezinto ezingaphili ongabizi, futhi ukusetshenziswa kwawo ekuhlanzeni amaprotheni ngokuyinhloko kusekelwe ekhonweni lawo lokufaka imvula yamaprotheni.

 

2.1 Ukuna kukasawoti

 

I-Ammonium sulfate yandisa amandla e-ion esixazululo, inciphisa amandla anyanyekayo phakathi kwama-molecule amaprotheni, futhi ikhuthaze ukuhlangana kanye nemvula yama-molecule amaprotheni. Le ndlela ibizwa ngokuthi 'izulu likasawoti'.

 

2.2 Imvula ekhethiwe

 

Ukuncibilika kwamaprotheni ahlukene kuyehluka ezindaweni ezihlukene ze-ammonium sulfate, okuvumela ukuna okukhethekile kwamaprotheni ahlosiwe ngokukhuphula kancane kancane ukugcwala kwe-ammonium sulfate.

 

2.3 Ukuvikela umsebenzi wamaprotheni

 

I-Ammonium sulfate ibhekwa njengento engasebenzi engajwayele ukusabela okungaqondile ngamaprotheni, okusiza ukuvikela ukwakheka kwemvelo nokusebenza kwamaprotheni.

 

3. Izinyathelo zenqubo yokuhlanza i-ammonium sulfate

 

3.1 Ukuhlukaniswa kweseli

 

Okokuqala, kuyadingeka ukudiliza amaseli futhi ukhulule amaprotheni e-intracellular ngokusebenzisa izindlela zomzimba noma zamakhemikhali.

 

3.2 Ukuhlolwa kokuqala kanye ne-sedimentation

 

Engeza i-ammonium sulfate ekugxiliseni okuthile ukuze uphuze iningi lamaprotheni futhi ukhiphe ukungcola.

 

3.3 Ihlola kabusha imvula

 

Khulisa kancane kancane ukugcwala kwe-ammonium sulfate futhi unciphise amaprotheni okuhlosiwe.

 

3.4 Ukumiswa nokugeza

 

Misa kabusha iphrotheni enile ku-buffer futhi ugeze ukuze ususe ukungcola okusele.

 

3.5 Ukuhlanzwa okwengeziwe

 

Uma kudingekile, iphrotheni ingase idinge ukuphinde ihlanzwe nge-ion exchange chromatography, i-gel filtration chromatography noma i-affinity chromatography.

 

4. Izinzuzo kanye nokubi ukuhlanzwa kwe-ammonium sulfate

 

Inzuzo

 

Ukusebenza kwezindleko: I-Ammonium sulfate ayibizi futhi ifanele ukusetshenziswa kwezinga elikhulu.

Kulula ukusebenza: Inqubo yokwehlisa i-sedimentation ilula futhi ayidingi imishini eyinkimbinkimbi.

Umsebenzi wokuvikela: unomthelela omncane esakhiweni semvelo kanye nokusebenza kwamaprotheni.

 

Ukushoda

 

Ukulungiswa okulinganiselwe: Kunzima ukuhlukanisa amaprotheni anezakhiwo ezifanayo.

Ukungcola okungenzeka: I-Ammonium sulfate ngokwayo ingase yethule ukungcola, okudingeka kususwe ngezinyathelo ezilandelayo.

 

5.Izitayela zesikhathi esizayo zokuhlanzwa kwe-ammonium sulfate

 

Ngokujula kocwaningo lwamaprotheni, izidingo zobuchwepheshe bokuhlanza amaprotheni nazo zikhula njalo. Ukuhlanzwa kwe-ammonium sulfate esikhathini esizayo kungase kuhlanganiswe nobuchwepheshe besimanje, obufana:

 

Ubuchwepheshe be-chromatography bokucaca okuphezulu: buthuthukisa ukulungiswa kokuhlanza futhi buhlukanise amaprotheni amaningi.

 

I-Automation kanye ne-miniaturization: ukuthuthukisa ukusebenza kahle nokunemba.

 

I-Green chemistry: ukunciphisa ukusetshenziswa kwamakhemikhali kanye nokungcoliswa kwemvelo.

 

I-Epilogue

 

I-Ammonium sulfate, njenge-precipitant yendabuko yamaprotheni, idlala indima engenakushintshwa emkhakheni wokuhlanza amaprotheni. Ukusebenza kahle kwayo kwezindleko, ukusebenza kalula, kanye nokuvikelwa komsebenzi wamaprotheni kuyenze yasetshenziswa kabanzi ezinqubweni zokuhlanza amaprotheni. Ngokuthuthuka kwesayensi nobuchwepheshe, ubuchwepheshe bokuhlanza i-ammonium sulfate kulindeleke ukuthi buhlanganiswe nobuchwepheshe besimanje ukuze kuthuthukiswe ukusebenza kahle nokusebenza kahle kokuhlanzwa kwamaprotheni, kunikeze ukwesekwa okuqinile kocwaningo lwamaprotheni kanye nokusetshenziswa.