Ob txoj hauv kev rau cleaving peptides los ntawm cov khoom -theem resins
Ib txoj hauv kev rau kev tshem cov polypeptides los ntawm cov khoom -theem resins
Khoom theem Fmoc Tom qab chemically synthesizing peptides, muaj ntau txoj kev ntawm cleavage. Tam sim no peb yuav qhia ob hom yooj yim uas siv los ntawm txhua tus.
1. Strong acid method (feem ntau siv TFA)
Muab cov pob zeb qhuav tso rau hauv lub khob shaker thiab ntxiv qhov tsim nyog ntawm scavenger (Scavengers) Cleaving reagent trifluoroacetic acid (TFA), 10-25ml / g Resin, npog, thiab co intermittently ntawm chav tsev kub. Lim cov resin nrog TFA tshiab Ntxuav cov cob 2-3 zaug, muab cov filtrate, thiab dropwise ntxiv 8-10 npaug ntawm qhov ntim ntawm glacial ether. Qee lub sij hawm ib feem ntawm nws tuaj yeem raug steamed tawm TFA A crude peptide precipitate tau txais. Diethyl ether tuaj yeem ua kom txias nrog dej khov los txhim kho nag lossis daus. Thaum kawg, sau cov precipitated crude peptide.
Siv tau resin:King resin los yog Rink Amide Resin
2. Txoj kev tsis muaj kua qaub (diluted TFA lossis acetic acid)
1% TFA:
yuav 1 g Cov polypeptide- dai resin tau ntxiv rau cov tshuaj uas muaj 1% TFA Lub dichloromethane kua iav shake flask tau kaw thiab co rau 2 feeb. Siv nitrogen los nchuav cov lim rau hauv lwm lub khob methanol tshiab uas muaj 10% pyridine. Rov ua cov haujlwm saum toj no 10 zaug. Siv 30 ml DCM, 30ml MeOH Ntxuav cov peptides uas tseem tshuav rau ntawm cov cob peb zaug ua ntu zus. los ntawm TLC lossis HPLC Saib xyuas cov khoom seem. Muab tag nrho cov filtrate thiab evaporate cov kuab tshuaj nyob rau hauv lub siab txo kom tawm 5% ntim. ntxiv 40 ml dej, thiab dej khov - txias qhov sib tov los pab nag lossis daus. Tom qab pom, ntxuav cov khoom nrog dej huv peb zaug thiab muab tso rau hauv lub tshuab nqus tsev ziab kom siv. KOH lossis P2O5 Cia kom qhuav.
dilute acetic acid:
siv AcOH / TFE / DCM (2: 2: 6) Kho cov peptides dai ntawm cov resin nrog cov tshuaj txiav rau 2 teev, lim cov resin, thiab ntxuav cov resin peb zaug nrog kev txiav. Ntxiv 15 npaug ntawm qhov ntim ntawm n-hexane mus rau qhov sib xyaw filtrate, thiab rotary evaporate kom tshem tawm cov acetic acid ntau dhau. Cov txheej txheem rov qab Peptide tau piav qhia hauv qab no.
Siv tau resin:2- CTC cob