Yuav ua li cas yog PTFE rhaub siv nyob rau hauv Peptide Synthesis thiab Oligonucleotide Production?

Peptide thiab oligonucleotide synthesis cycles xaus nrog cov kauj ruam cleavage uas tshem tawm qhov tshwm sim ntawm cov resin ntawm trifluoroacetic acid (TFA) los yog lwm yam muaj zog acids. Txhawm rau kom muaj txiaj ntsig zoo, qhov sib xyaw corrosive no yuav tsum tau ua kom sov thiab qhov no yuav tsum muaj lub tshuab cua sov uas tuaj yeem tiv taus cov kua qaub thiab tsis kis tus nqi siab - txiaj ntsig. Rau kev kho mob peptides thiab hluavtaws oligonucleotides, purity xav tau siab heev thiab xaiv cov khoom siv cua sov ua ib qho tseem ceeb ntawm cov txheej txheem.

Cleavage thiab Deprotection Kauj Ruam hauv Khoom Phaj Synthesis
Lub hom phiaj molecule yog sib sau ua ke ntawm ib qho insoluble resin kev them nyiaj yug los ntawm cov theem peptide synthesis (SPPS) thiab khoom theem oligonucleotide synthesis. Tag nrho cov kab ke tau sib sau ua ke thiab tom qab ntawd cov txiaj ntsig yuav tsum tau cleaved los ntawm cov resin. Tib lub sijhawm, sab saw tiv thaiv pawg (rau peptides) lossis hauv paus / acid-labile tiv thaiv pawg (rau oligonucleotides) raug tshem tawm.

Cov cleavage cocktail li niaj zaus rau peptide synthesis yog trifluoroacetic acid (TFA) uas yog lub ntsiab cleavage agent, feem ntau ua ke nrog scavengers xws li triisopropylsilane (TIS), dej los yog 1,2-ethanedithiol (EDT). Rau oligonucleotide synthesis, deprotection tuaj yeem ua tau siv cov concentrated aqueous ammonia los yog, hauv qee qhov ultra- cov tswv yim me, nrog TFA-muaj cov kev daws teeb meem.

Feem ntau, cov txheej txheem cleavage yog ua nyob rau ntawm qhov kub thiab txias, feem ntau ntawm 30℃thiab 60 degree, thiaj li yuav ua kom tiav kev tso tawm ntawm cov khoom nyob rau hauv ib lub sij hawm (1-4 teev). Cov kub kub ceev ceev cleavage thiab deprotection, tab sis lawv kuj nce qhov muaj peev xwm ntawm cov khoom deterioration yog tias tsis tswj kom zoo. Cov cua sov yuav tsum yog homogeneous, rov tsim dua thiab tsis muaj kab mob.

TFA-Cov kev daws teeb meem - Qhov Kev Sib Tw Corrosive
Trifluoroacetic acid (pKa ~ 0.23) yog ib qho muaj zog organic acid uas muaj peev xwm ua kom yaj ntau cov hlau thiab ua rau cov khoom siv hluav taws xob raug puas tsuaj. TFA vapor kuj yog corrosive heev ntawm qhov kub thiab txias. Hlau immersion heaters (titanium, stainless hlau 316L, los yog Hastelloy) tuaj yeem tsim pitting, intergranular corrosion, los yog nto roughening tom qab raug kub TFA tshaj ob peb cleavage cycles.

Corrosion tuaj yeem ua kom paub cov hlau ions (hlau, chromium, npib tsib xee, titanium, thiab lwm yam) rau hauv cov tshuaj cleavage. Cov hlau ions tuaj yeem tsim cov complexes nrog cov khoom peptide lossis oligonucleotide, uas ua rau:

Txo qhov purity ntsuas los ntawm HPLC lossis LC-MS

Kev ua lom neeg lom neeg uas tsis tuaj yeem pom

Cov hlau tsis huv uas nyuaj rau kev tshem tawm hauv cov khoom lyophilised kawg

Ntxiv mus, hlau ions tuaj yeem ua rau cov txheej txheem sab nraud xws li oxidation ntawm methionine lossis tryptophan residues hauv peptides, lossis depurination hauv oligonucleotides. Cov tshuaj no tsis tso cai rau GMP Qib tshuaj tshuaj.

Yuav ua li cas PTFE Heaters Kho qhov Cleavage Cua Sov Teeb Meem
Polytetrafluoroethylene (PTFE) immersion heaters yog teev rau daim ntawv thov no, raws li PTFE yog lig resistant rau trifluoroacetic acid txawm nyob rau hauv siab kub. Pua pua ntawm cleavage cycles ua rau tsis muaj tshuaj tua kab mob, tsis o los yog degradation nto. Lub PTFE rhaub peptide synthesis oligonucleotide tsim muaj ib tug huv si, inert cua sov qhov chaw ncaj qha nyob rau hauv lub cleavage hlab los yog nyob rau hauv lub pre- cua sov reservoir rau lub TFA cocktail.

Tag nrho Resistance rau TFA thiab Scavengers
PTFE ruaj khov rau TFA, TIS, EDT, phenol, thioanisole thiab lwm yam txheej txheem cleavage cocktail Cheebtsam. Tib yam tshuaj inertness kuj tau pom rau oligonucleotide deprotection agents, xws li concentrated ammonia lossis methylamine. Immersion rhaub ua los ntawm PTFE tuaj yeem dipped ncaj qha mus rau hauv cov tshuaj cleavage yam tsis muaj corrosion los yog leaching ntawm ions.

Tsis yog -Ntoo Nto, Tsis muaj qhov seem tsim -Up
Lub du, tsis yog -stick nto ntawm PTFE kuj yog resistant rau tsub zuj zuj ntawm peptide los yog oligonucleotide residues, cleaved resin fragments, los yog scavenger los ntawm - khoom. Kev yaug cov khoom siv nrog cov kuab tshuaj xws li dichloromethane, acetonitrile lossis dej yog yooj yim. Qhov no tso cai rau kev tu kom yooj yim dua, tiv thaiv tus ntoo khaub lig- sib kis ntawm cov khoom sib txawv sib txawv, thiab ua kom yooj yim rau kev siv tau rau GMP ntau lawm.

Kev tswj qhov kub thiab txias rau qhov siab tshaj plaws
Lub PTFE immersion rhaub feem ntau yog txuas nrog PID maub los thiab lub ntsuas kub (PT100 lossis thermocouple) ncaj qha kov cov tshuaj cleavage. Qhov qis thermal loj ntawm PTFE sheath tso cai rau cov tshuaj tiv thaiv sai rau kev hloov pauv. Ib qho cleavage cua sov profile tuaj yeem yog:

Ramp los ntawm ambient mus rau 40℃(tshaj 10 min)

Tuav ntawm 40℃± 0.5℃rau 90 feeb

Tom qab ntawd txias rau 25℃thiab cov khoom precipitated thiab lim

Qhov kev tswj xyuas meej no tau lees paub cov khoom lag luam siab tshaj plaws thiab, tib lub sijhawm, tiv thaiv kev tawm tsam xws li aspartimide ntau lawm hauv peptides lossis cleavage ntawm phosphodiester bonds hauv oligonucleotides.

Daim ntawv ceeb toom: Sizing lub PTFE rhaub rau Common Cleavage Volumes
Daim ntawv ceeb toom: Rau peptide synthesis ntawm lub ntsuas ntsuas (0.1 txog 10 mmol resin load), cleavage kom muaj nuj nqis feem ntau yog 5 mL txog 500 mL. Qhov tsim nyog yog lub rhaub dej kub ntawm PTFE kev tsim kho, 50 mus rau 200 W. Lub watt ceev ntawm lub rhaub yuav tsum tau khaws cia qis (tsawg dua los yog sib npaug li 10 W / cm^2) los tiv thaiv lub zos boiling ntawm TFA (boiling point ~ 72℃). Automated synthesisers lossis pilot-scale reactors (1-50 L cleavage ntim) tuaj yeem muab nrog ntau lub tshuab cua sov PTFE lossis ib lub rhaub loj dua (500-3000 W). Lub geometry ntawm lub rhaub (ncaj, L-shaped los yog coil) yuav tsum raug xaiv kom haum rau hauv lub nkoj cleavage yam tsis muaj kev cuam tshuam nrog kev sib nqus nplawm lossis kev kub ntxhov.

Nyob rau hauv automated synthesisers PTFE heaters feem ntau muaj nyob rau hauv lub cleavage module raws li ib tug pre-heater rau TFA reagent kwj. Cov kua qaub yog khiav los ntawm ib qho me me PTFE hauv -kab cua sov ua ntej nws mus txog cov hlab ntsha. Qhov no tau lees tias qhov cleavage cocktail yog ntawm lub hom phiaj kub tam sim ntawd thaum nws hits cov resin.

Purity of the Final Product is depend on Contamination-Tswv Cua Dawb
Ntshiab yuav tsum yog tag nrho cov seem uas kov cov cleavage tov kom qhov kawg tshwm sim ntshiab. Lub PTFE rhaub peptide synthesis oligonucleotide system lav tias tsis muaj hlau ions, extractables los yog hais tau qhia thaum lub sij hawm tseem ceeb deprotection thiab cleavage txheej txheem. Qhov no yog qhov tseem ceeb tshwj xeeb rau kev kho peptides thiab oligonucleotides npaj rau tib neeg siv, rau cov tshuaj pharmacopoeias (xws li USP.<232>, ICH Q3D) teeb tsa siab pib rau lub hauv paus impurities.

Xaus: Leading Advanced Biopharmaceutical Manufacturing
PTFE cua sov yog ib qho tseem ceeb ua kom muaj kev tiv thaiv rau qhov kawg, theem tseem ceeb hauv peptide thiab oligonucleotide ntau lawm, kom ntseeg tau tawm los thiab purity. Lub tshuab cua sov muab cov tshuaj lom neeg tsis zoo, tsis yog- paug, thiab hloov kho qhov kub ntawm qhov muaj tseeb thaum muaj TFA kub thiab lwm yam kev ua phem cleavage agents, ua rau cov kws tshuaj hluavtaws los ua kom zoo dua qhov kev sib cais tsis muaj kev cuam tshuam qhov zoo ntawm qhov tshwm sim.

Advanced biopharmaceutical manufacturing xav tau siab -purity cua sov daws. Qhov kev thov nce ntxiv rau cov peptides hluavtaws (xws li, GLP-1 agonists) thiab kho cov tshuaj oligonucleotides (piv txwv li, antisense thiab siRNA tshuaj) yuav txhawb nqa kev siv PTFE immersion heaters rau cleavage thiab deprotection cov kauj ruam raws li kev xyaum hauv kev tshawb fawb thiab GMP ntau lawm chaw.