Iingxaki kunye nezisombululo ze-peptide synthesis ende

Kuphando lwebhayoloji, iipolypeptides ezinolandelelwano olude zihlala zisetyenziswa.Kwiipeptides ezingaphezulu kwe-60 amino acids kulandelelwano, i-gene expression kunye ne-SDS-PAGE zisetyenziswa ngokubanzi ukuzifumana.Nangona kunjalo, le ndlela ithatha ixesha elide kwaye umphumo wokugqibela wokuhlukana kwemveliso awulungile.

Imingeni kunye nezisombululo ze-peptide synthesis ende

Kwi-synthesis yeepeptides ezinde, sihlala sijongene nengxaki, oko kukuthi, isithintelo esinqabileyo sokuphendula kwe-condensation sinyuka ngokunyuka kolandelelwano kwi-synthesis, kwaye ixesha lokuphendula kufuneka lihlengahlengiswe ukwenza ukuphendula kuphelele.Nangona kunjalo, ixesha lokuphendula ixesha elide, iziphumo ebezingalindelekanga ezininzi ziveliswa, kwaye inxalenye yepeptide ekujoliswe kuyo yenziwa.Iintsalela ezinjalo - amakhonkco e-peptide anqongopheleyo bubumdaka obuphambili obuveliswe kwi-peptide synthesis ende.Ke ngoko, ekudityanisweni kwepeptide ende, eyona ngxaki iphambili ekufuneka siyoyise kukuphonononga iimeko zokusabela ezikumgangatho ophezulu kunye neendlela zokusabela, ukuze senze i-amino acid yokujikwa kwe-condensation iphelele kwaye iphelele.Ukongeza, nciphisa ixesha lokuphendula, kuba ixesha lokuphendula ixesha elide, ukusabela okungalawulekiyo kwecala, kokukhona kuntsonkothileyo kweemveliso.Ngoko ke, ezi ngongoma zintathu zilandelayo zishwankathelwa:

I-Microwave synthesis ingasetyenziswa: Kwezinye i-amino acids ezidityaniswe kwinkqubo yokudibanisa ekungekho lula ukudibanisa, i-microwave synthesis ingasetyenziswa.Le ndlela ineziphumo eziphawulekayo, kwaye inciphisa kakhulu ixesha lokuphendula, kwaye inciphisa ukubunjwa kwezinto ezimbini eziphambili zeemveliso.

Indlela yokudibanisa iqhekeza ingasetyenziswa: Xa ezinye iipeptides kunzima ukudityaniswa ngeendlela eziqhelekileyo zokudityaniswa kwaye kungekho lula ukucocwa, sinokuthatha yonke i-condensation yee-amino acids ezininzi kwicandelo elithile lepeptide ukuya kwikhonkco lepeptide lilonke.Le ndlela inokusombulula iingxaki ezininzi kwi-synthesis.

I-Acylhydrazide synthesis ingasetyenziswa: i-acylhydrazide synthesis ye-peptides yindlela ye-slid-phase synthesis ye-N-terminal Cys peptide kunye ne-C-terminal polypeptide hydrazide ikhemikhali yokusabela okukhethiweyo phakathi kokubunjwa kweebhondi ze-amide ukufezekisa indlela yokudibanisa i-peptide.Ngokusekwe kwindawo yeCys kwikhonkco lepeptide, le ndlela yohlulahlula yonke ikhonkco lepeptide ibe lulandelelwano oluninzi kwaye luyidibanise ngokulandelelana.Okokugqibela, i-peptide ekujoliswe kuyo ifunyanwa ngokusabela kolwelo-kwinqanaba lokujiya.Le ndlela ayinciphisi kakhulu kuphela ixesha lokudibanisa i-peptide ende, kodwa ikwanyusa kakhulu ukucoceka kwemveliso yokugqibela.

Ukucocwa kwepeptide ende

Ubunjani beepeptides ezinde ngokungaphephekiyo bukhokelela kumalungu antsonkothileyo eepeptide ezikrwada.Ke ngoko, kungumceli mngeni ukuhlambulula iipeptide ezinde ngeHPLC.uchungechunge lwe-amyloid lwenkqubo yokucoca i-polypeptide, ifunxa amava amaninzi kwaye isetyenziswe ngempumelelo ekucoceni i-peptide ende.Ngokwamkela izixhobo ezitsha, ukuxuba iinkqubo ezininzi zokucoca, ukuhlukana ngokuphindaphindiweyo kunye nezinye iindlela zamava, izinga lokuphumelela lokuhlanjululwa kwe-peptide elide liphuculwe kakhulu.