Muaj - nplai downstream purification ntawm bacteriopages - 'clubification' thiab 'ultrafiltration'
Bacteriophages, Kuj muaj cov lus hu ua Phages, yog cov kab mob uas kis tau tus kab mob. Cov phage tsis muaj sia nyob ntawm nws tus kheej; Nws yuav tsum mob siab rau cov kab mob tus tswv tsev kom cov me nyuam, thaum kawg ua rau tus kab mob tsis pom zoo. Vim tias lawv cov khoom muaj txiaj ntsig zoo, cov ncauj lus tuaj yeem siv tau kho mob rau cov kab mob cim thiab ntaus, nrog rau kev kho mob cov kab mob refractory.
Qhov no yog schematic daim duab ntawm ib tug bacteriophage qauv (duab duab: Is Taws Nem).
Nws yog kwv yees hais tias cov nroj tsuag kab mob ua rau muaj ntau dua 30% ntawm cov kab mob thoob ntiaj teb cog qoob loo txhua xyoo. Ntawm ntau cov pathogens, cov kab mob kab mob yog qhov tshwj xeeb nyuaj rau kev tswj hwm. Cov txheej txheem tswj ib txwm muaj feem ntau ntawm cov tshuaj tua kab mob thiab tooj liab {}}} bathed cov neeg sawv cev. Txawm li cas los xij, cov tshuaj tua kab mob ntau dhau lawm tau ua rau nce cov tshuaj tiv thaiv loj, thaum lub sijhawm ntev siv ntawm kev sib tw ib puag ncig, ntawm kev pheej hmoo kev pheej hmoo rau tib neeg thiab tsiaj.
Vim tias cov bacteriophagent muaj cov kab mob muaj txiaj ntsig zoo, lawv tuaj yeem xaiv cov kab mob tua kab mob uas tsis muaj cov microbes lossis lwm yam host hlwb. Yog li, cov phooj ywg tuaj yeem ua lwm yam tshuaj tua kab mob thiab tooj liab -} buil} }ອກ. Cov tshuaj lom phas, pathogens tuaj yeem tshem tawm tau zoo thaum txo cov tshuaj tua kab mob thiab tooj liab.
Nrog kev tshawb fawb tas mus li thiab kev tshawb fawb thev naus laus zis thiab kev tshawb nrhiav tob hauv bacteriopcages, lub zeem muag rau kev siv superbugs tau ua cog lus ntau zuj zus. Nyob rau yav tom ntej, page kev kho mob yuav tsum tau dhau los ua ib qho ntawm cov kev daws qhov tseem ceeb los tiv thaiv kev tiv thaiv. Los ntawm kev tshawb fawb txuas ntxiv thiab kev tshawb nrhiav, phage kev kho mob tuaj yeem tawm los ua cov quab yuam loj hauv kev kho mob antimicrobial, ua rau tib neeg kev noj qab haus huv.
Txawm li cas los xij, kom muaj kev nyab xeeb thiab tswj hwm bacterongopcages rau tib neeg lub cev -}- ib qho kev sib tw tseem ceeb:Yuav ua li cas kom tau Ultra - Cov Phot Eje Ejections.
Phoj lysate yog ib txoj kev sib xyaw ua ke uas, ntxiv rau cov kab mob loj xws li: cov neeg tuav pov hwm, thiab cov tswv cuab {}} Lwm yam impurities); endotoxins xws li Lipopolysaccharides (LPS) (txheej txheem -} lwm yam cuam tshuam); Thiab khoom {{- muaj feem cuam tshuam cov impurities suav nrog khoob capsids thiab tawg tails.
Yog li ntawd, lub hom phiaj ntawm kev ua kom huv tsis yog tsuas yog ua tiav cov phoj siab heev tab sis kuj tau ua kom txo qis {{}}} rau cov qauv kev ua haujlwm tsawg raws li cov chaw muag tshuaj tau teev tseg.
Lub zog ua kom ruaj khov thiab ntsuas cov txheej txheem phage pho ntsuas feem ntau ua raws cov hauv paus ua haujlwm hauv qab no thiab tuaj yeem faib ua cov kauj ruam hauv qab no:
Txheej txheem hauv qab ntawm bacteriopcages
Kev Hais Qhia - Tshem Tawm ntawm macroscopic impurities
Nyob rau hauv thawj theem ntawm bacteriophage purification, lub hom phiaj ntawm cov theem qhia meej yog kom tshem tawm cov kab mob hlwb thiab cov kab mob cellular los ntawm cov lysate crude. Lub hom phiaj tseem ceeb ntawm cov kauj ruam no yog los muab kev huv si rau poob qis cov chav nyob lossis lub chav ntim khoom sib cais, txo qis cov khoom siv ntawm cov khoom siv. Qhov no zoo tiv thaiv clogging hauv cov chaw ua haujlwm lim dej tom qab, ua haujlwm ruaj khov thiab cov txheej txheem siab thoob plaws thoob plaws hauv kev ua haujlwm lim.
Hauv paus chiv keeb bacterongage, kev piav qhia tshwj xeeb yog li ntawm 0.8μm, 0.24μm}}}}}}}}}}}}}}}} Txawm hais tias paub tab thiab ntseeg tau, txoj hauv kev no cuam tshuam, yog lub sijhawm {{{{{}} Kev ua kom zoo dua nyob rau hauv chav ua kom zoo nyob rau hauv tag nrho cov txiaj ntsig thiab kev ua tau zoo.
Hloov cov mult -} theem pom kev lim dej nrog aMicrofiltration Tshuaj Ntsiavtuaj yeem ua kom yooj yim yooj yim thiab ua kom cov txheej txheem. Tshwj xeeb, tom qab tshem tawm cov kev khib cell feem ntau dhau los ntawm qis- kev lysate tuaj yeem ua ncaj qha los ntawmtangential khiav pom kev (TFF)Siv microfiltration cantsules nrog txhais lub qab loj me (0.45μm lossis 0.22μm). Qhov no ua rau kev raug tshem tawm zoo ntawm kev cais tawm thiab muaj txiaj ntsig zoo ntawm cov ntsiab lus hauv cov lus hauv aIb Leeg Ib Leeg, ua rau muaj kev sib xyaw ua ke ntawm peb ntu ntawm ib qho.
Qhov kev hloov tshiab no tsis yog txo qis lub sijhawm ua haujlwm tab sis kuj txo cov qauv kev pheej hmoo los ntawm ntau cov kauj ruam pom kev, yog li kev txhim kho page rov qab. Lub caij no, cov haujlwm ua haujlwm txaus ntawm cov haujlwm ua haujlwm mitigates membrane fouling, txhim kho kev tawm tsam, thiab cov peev txheej loj.
Yog li ntawd, tau txais ib qhoKev sib xyaw ua ke lub tswv yim ua ke qis qis -} ceev Centrifugation thiab microfililation capsulesSawv cev txoj hauv kev ua tau zoo los txhim kho kev ua tau zoo thiab raug nqi {{0}-} nplai bacteriopage ntau lawm. Guidling Technology's Microfilidation Ciav tshuaj yuav txo qis kev noj cov tshuaj lom rau hauv qab20 ntu, siab sib ntsib cov kev xav tau ntawm txos ultrafiltration thiab chromatography steption.
Kev ntes thiab concentration - thawj kev ua kom huv thiab ntim txo
Thaum lub sijhawmntes thiab siabTheem ntawm bacteriophage purification, lub hom phiaj tseem ceeb yog kom tau txais txiaj ntsig zoo ntawm cov lus qhia kom meej thaum ib txhij ua tiav cov khoom lag luam tseem ceeb thiab kev sib pauv ntas. Cov kauj ruam no feem ntau nyob ntawmTangential khiav pom kev (TFF)thev naus laus zis.
Lub hauv paus ntsiab lus ntawm Tff dag nyob rau hauv kev siv ultrafiltration membranes nrog qhia meej molecular hnyav txiav {}} Offs (feem ntau 100 lossis 300kda). Cov me me molecular impurities xws li residual kab lis kev cai xov xwm, thiab cov protein ntau, thiab cov khoom lag luam me me dhau los ntawm cov membrane pores, thaum cov ncauj yog khaws cia vim yogQhov loj me tsis suav nrog cuam tshuam, ua kom lawv sib cais ua tau zoo thiab txhawb nqa.
Tsis pub dhau ib tee,UltraFiltration homtau ua haujlwm kom ua tiav cov ntim kev pom zoo, thaum hloov musDiafiltration homTso cai rau kev sib pauv tsis sib pauv, yog li tsim tsim kom haum cov kws kho hlwb ua rau cov kauj ruam tom ntej xws li kev kho mob enzymatic.
Cov thev naus laus zis no muaj cov txiaj ntsig zoo sib xyaw ntawmUa hauj lwm siabthiabCov Toadity Zoo Tshaj, ua rau nws zoo tagnrho rau loj - nplai ntau lawm. Raws li Guidlingtechnology, UltraFiltration tsiav tshuaj yuav ua tiav ib qhoPage Rov qab rov qab tus nqi ntawm 90-95%, nyob ntawm cov khoom siv tshwj xeeb tau ua tiav.
Kev nqus kom huv - tsom rau kev tshem tawm ntawm qhov tseem ceeb heev
Hauv bacteriophage purification,sib sib zog nqusyog cov kauj ruam tseem ceeb uas txiav txim siab cov khoom lag luam kawg. Nws thawj lub hom phiaj yog tusKev tshem tawm tshwj xeeb ntawm qhov tseem ceeb impurities, xws li endotoxins. Coj kev cai qubCSCL Density Gradient Centrifugation{{0- Vim nws cov tshuaj lom thiab tsis muaj kev sib zog ntawm cov txheej txheem. Tam sim no txoj kev tsom mus rauchromatography -} Raws technologies, nrog kev sib koom ua ke tshiabenzyme pretreatment.
Ib txoj kev ua tiav qib siab ua keanion txauv cov chromatography (AEC)nrogAlkaline Phosphatase (AP) Pretreatmenttau tsim. Ob kab lus thiab Lipopolysaccharides (LPS) Nqa kev sib tw tsis zoo rau cov chaw tsis zoo, kev ua kom muaj kev sib xyaw ua ke. Los ntawm kev qhia txog ap pretreatment ua ntej AEC loading, tus enzyme tshwj xeeb tshem tawm cov kab tsib ntawm LPS molecules, txo qis lawv tus nqi tsis zoo. Qhov no zoo ua tsis muaj zog lawv ib qho kev sib txawv rau lub anion sib pauv nruab nrab.
Cov ntaub ntawv sim qhia tias kho cov qauv nrog20 u / mltaug qab los ntawm kev huv sivQuaternary Amine (Q) Ligand Membrane Adsorbers lossis monolithic Txhuatuaj yeem ua tiav98.8% Tshem Tawm Kev Tshem Tawm, thaum tswj cov nqi pharess zoo heev. Txoj kev ua tiav no tau daws tau qhov ntev - sawv qhov teeb meem ntawm endoxin co- adsorption thaum page purification
Ua kom zoo dua Polishing - Kawg ua kom zoo nkauj thiab cov qauv
Thaum kawgPolishing theemntawm bacteriophage purification, lub hom phiaj tseem ceeb yog kom ua tiav kev coj dawb huv thiab kev tsim qauv tsim. Qhov no suav nrog kev tshem tawm cov kab tseem ceeb, tshem tawm cov qauv phosphatase (xws li alkaline phosphatase), thiab kev sib pauv khoom rau hauv kev ua kom tiav.
Qhov no ib txwm ua tiav los ntawmtheem nrab diafiltration, ib txoj kev pov thawj thiab muaj txiaj ntsig zoo uas ua rau cov kev tshem tawm me me ntawm - molecule cov khoom tsis huv thiab meej muffer sib pauv.
Txhawm rau txhim kho cov khoom lag luam purity,hydrogen - bond cuam tshuam cov chratinaphlossisSib xyaw - hom chromatography (MMC)tuaj yeem qhia raws li aKauj Ruam Zaum Kawg Kauj RuamCov. Cov tswv yim no siv ntau lub tshuab sib cais kom tshem tawm cov kab nrog cov khoom siv dag zog zoo ib yam li cov hom phiaj pharic. Qhov tshwm sim yog ib qho bacteriophage nquag muaj cov tshuaj ntsuab (API) uas tau raws li cov qauv nruj ua kom tau rauTxhaj Tshuaj - qeb raws cov qauv.
Los ntawm kev xaiv kev sib xyaw ntawm cov microfiltration thiab ultrafiltration membrane uas tuaj yeem ua tiav cov khoom siv tsis ntev los no yog kwv yees li ntawm 100-30 lmh. Piv nrog tsoos kev ua, txoj kev no tuaj yeem ua tau zoo ntawm cov txheej txheem uas yuav tsum muaj rau ob qho meej thiab ultrafiltration.
Tuaj txog::
Saavedra li al.,Scarectable Pauv Tawm Ntawm Bacteriophage Npaj (2025)
Lapras li al.,Rationalization ntawm cov txheej txheem lim tshuaj rau cov phoj ua khoom noj muaj txiaj ntsig (2024)