Izinkinga nezixazululo ze-peptide synthesis ende

Ocwaningweni lwebhayoloji, kuvame ukusetshenziswa ama-polypeptide anokulandelana okude.Kuma-peptide anama-amino acid angaphezu kuka-60 ngokulandelana, ukusho kofuzo kanye ne-SDS-PAGE kuvame ukusetshenziswa ukuze kutholakale.Nokho, le ndlela ithatha isikhathi eside futhi umphumela wokugcina wokuhlukanisa umkhiqizo awumuhle.

Izinselelo nezixazululo ze-peptide synthesis ende

Ekuhlanganisweni kwama-peptide amade, sihlala sibhekene nenkinga, okungukuthi, isithiyo esiqinile sokusabela kwe-condensation sikhuphuka ngokwanda kokulandelana kokuhlanganiswa, futhi isikhathi sokuphendula sidinga ukulungiswa ukuze kuphelele ukusabela.Kodwa-ke, uma isikhathi sokusabela sithatha isikhathi eside, kukhiqizwa imiphumela emibi kakhulu, futhi kwakheka ingxenye ye-peptide eqondiwe.Izinsalela ezinjalo - amaketanga e-peptide antula ukungcola okubalulekile okukhiqizwa ku-peptide synthesis ende.Ngakho-ke, ekuhlanganisweni kwe-peptide ende, inkinga ebalulekile okufanele siyinqobe ukuhlola izimo zokusabela zekhwalithi ephezulu kanye nezindlela zokusabela, ukuze senze ukusabela kokujiya kwe-amino acid kuphelele futhi kugcwale.Ukwengeza, nciphisa isikhathi sokuphendula, ngoba isikhathi eside sokuphendula, ukusabela okungalawuleki okungalawuleki, kuyinkimbinkimbi ngokwengeziwe imikhiqizo.Ngakho-ke, amaphuzu amathathu alandelayo afingqiwe:

I-Microwave synthesis ingasetshenziswa: Kwamanye ama-amino acid ahlangana nawo enqubweni yokuhlanganiswa okungelula ukuhlanganiswa, i-microwave synthesis ingasetshenziswa.Le ndlela inemiphumela ephawulekayo, futhi inciphisa kakhulu isikhathi sokuphendula, futhi inciphisa ukwakheka kwemikhiqizo emibili ebalulekile.

Indlela yokuhlanganisa izingcezu ingasetshenziswa: Uma amanye ama-peptide kunzima ukuhlanganiswa ngezindlela ezivamile zokuhlanganisa futhi kungelula ukuhlanzwa, singakwazi ukusebenzisa ukufiphala okuphelele kwama-amino acid amaningana engxenyeni ethile ye-peptide kuya kuchungechunge lwe-peptide lilonke.Le ndlela futhi ingaxazulula izinkinga eziningi ekuhlanganiseni.

I-Acylhydrazide synthesis ingasetshenziswa: I-acylhydrazide synthesis yama-peptides iyindlela ye-solid-phase synthesis ye-N-terminal Cys peptide kanye ne-C-terminal polypeptide hydrazide chemical selective reaction phakathi kokwakhiwa kwama-amide bond ukuze kuzuzwe indlela yokubopha i-peptide.Ngokusekelwe endaweni yama-Cys ochungechungeni lwe-peptide, le ndlela ihlukanisa lonke iketango le-peptide libe ngokulandelana okuningi futhi liwahlanganise ngokulandelana.Ekugcineni, i-peptide eqondiwe itholakala ngokusabela kokujiya kwesigaba se-liquid.Le ndlela ayigcini nje ukunciphisa kakhulu isikhathi sokuhlanganiswa kwe-peptide ende, kodwa futhi yandisa kakhulu ukuhlanzeka komkhiqizo wokugcina.

Ukuhlanzwa kwe-peptide ende

Ukukhethekile kwama-peptide amade kuholela nakanjani ezingxenyeni eziyinkimbinkimbi zama-peptide aluhlaza.Ngakho-ke, futhi kuyinselele ukuhlanza ama-peptide amade nge-HPLC.uchungechunge lwe-amyloid lwenqubo yokuhlanza i-polypeptide, emunca ulwazi oluningi futhi lusetshenziswe ngempumelelo ekuhlanzweni kwe-peptide ende.Ngokwamukela imishini emisha, ukuxuba izinhlelo eziningi zokuhlanza, ukuhlukaniswa okuphindaphindiwe nezinye izindlela zokuhlangenwe nakho, izinga lokuphumelela lokuhlanzwa kwe-peptide ende lathuthukiswa kakhulu.