300kd Hollow Fiber Ultrafiltration: Kev Tshawb Fawb Txog Kev Ua Haujlwm Siab -Kev Ua Haujlwm Cuam Tshuam Thiab Cov Txheej Txheem Ua Kom Zoo Rau Ko Taw-thiab- Kab Mob Hauv Qhov Ncauj O/A Tus Kab Mob
Sep 03, 2026| Taw qhia
Ko taw-thiab- kab mob qhov ncauj (FMD) yog ib qho mob hnyav, febrile, thiab kis tau tus tsiaj kis tau zoo heev los ntawm ko taw- thiab- kab mob qhov ncauj (FMDV). Tus kab mob no cuam tshuam rau cloven-cov tsiaj nyeg, nrog ntau dua 70 hom kab mob. Nws kis tau sai heev thiab muaj tus kab mob siab, feem ntau cuam tshuam rau cov tsiaj txhu loj xws li npua, yaj, thiab nyuj, qhov xwm txheej tuaj yeem ncav cuag 100%. FMDV belongs rau lub genusKo taw-thiab-Mouthvirusntawm tsev neeg MicroRNAviridae. Cov khoom yog spherical, kwv yees li 27-30 nm inch, nrog sedimentation coefficient ntawm 146S rau cov kab mob tsis zoo. Tam sim no, muaj ntau yam tshuaj tiv thaiv FMD, suav nrog cov tshuaj tiv thaiv kab mob muaj sia nyob, tshuaj tiv thaiv kab mob muaj sia nyob, tshuaj tiv thaiv tsis muaj zog, tshuaj tiv thaiv nucleic acid, tshuaj tiv thaiv peptide hluavtaws, thiab tshuaj tiv thaiv subunit. Cov tshuaj tiv thaiv FMD uas tsis muaj zog feem ntau muaj cov kev mob tshwm sim tsawg dua hauv cov tsiaj, thiab cov tshuaj tiv thaiv ntau yam muab kev tiv thaiv zoo dua, ua rau lawv yog qhov tseem ceeb ntawm kev tiv thaiv FMD hauv Suav teb.
Tangential Flow Ultrafiltration
Tam sim no, tangential flow ultrafiltration yog ib txoj hauv kev uas feem ntau siv purification thiab concentration nyob rau hauv qhov loj - nplai ntau lawm ntawm ko taw- thiab - qhov ncauj kab mob ntau yam tshuaj tiv thaiv tsis muaj zog. Txoj kev no tsis tsuas yog ua tiav qhov xav tau qhov xav tau, tab sis kuj tshem tawm ntau qhov txawv teb chaws impurities. Tangential flow ultrafiltration yog tsuas yog siv rau kev ua kom huv, concentration, thiab filtration (desalting thiab tsis hloov). Tsis tas li ntawd, nws tseem tuaj yeem siv rau kev qhia meej ntawm fermentation broth lossis cell kab lis kev cai nruab nrab siv tangential flow microfiltration, yog li hloov centrifugation lossis thawj qhov kev qhia meej.
Nyob rau hauv cov pa ntsug pom, loj molecules yuav aggregate rau ntawm daim nyias nyias, clogging lub lim. Tangential flow filtration, txawm li cas los xij, tso cai rau cov dej ntws mus rau qhov sib npaug ntawm daim nyias nyias, zam qhov teeb meem clogging. Thaum lub sij hawm tangential flow filtration, cov kuab tshuaj thiab cov khoom me dua li daim nyias nyias pore loj uas permeate los ntawm daim nyias nyias yog hu ua permeate, thaum cov khoom loj dua qhov pore loj yog khaws cia raws li cov concentrate. Ua ke nrog optimized ntxuav thiab pom cov txheej txheem, kev sib cais thiab purification ntawm molecules ntawm ntau qhov ntau thiab tsawg tuaj yeem ua tiav. Cov thev naus laus zis no muaj cov txiaj ntsig zoo xws li tus nqi qis, kev ua haujlwm me me, yooj yim tu thiab hloov pauv, siab membrane permeate txaus tus nqi, thiab kev ua haujlwm yooj yim, ua rau nws tsim tshwj xeeb rau qhov loj - nplai concentration thiab purification ntawm bioactive tshuaj.
Cov ntaub ntawv thiab cov txheej txheem
Hollow Fiber Kem:Molecular hnyav txiav tawm ntawm 300kd, membrane cheeb tsam ntawm 1600cm².
Khoom siv:5.6L txhua tus supernatant los ntawm FMDV hom O thiab hom A tom qab centrifugation thiab qhia meej.
Txheej txheem Parameter:Thaum lub sij hawm ultrafiltration concentration, lub zog shear raug hloov mus rau kwv yees li 3000⁻¹, thiab TMP tau tswj kom tsis txhob siab tshaj 10 psi. Thaum cov concentration ntau tshaj 11 zaug, cov qauv raug ntxuav nrog 6CV Buffer tov. Tom qab ntxuav, cov cua tsis muaj menyuam tau nkag mus rau hauv lub raj xa dej, thiab tag nrho cov qauv concentrated hauv lub raj yog txhaj rau hauv cov qauv concentration lub raj mis. Tom qab ntawd, sterile PBS tau ntxiv rau yaug lub raj kom txog thaum lub ntim kawg ntawm cov kua concentrated yog 500 mL, yog li txhim kho tag nrho cov nyiaj rov qab.
Cov txiaj ntsig thiab kev sib tham
Nyob rau hauv cov nqe lus ntawm thawj tus kab mob purification, ultrafiltration concentration ua tiav tus nqi tshem tawm ntau tshaj 90% rau impurities xws li cov proteins uas tsis muaj cov kab mob. Nyob rau hauv cov nqe lus ntawm ntim, nws muaj peev xwm ua tau siab concentrations ntawm 10-50 lub sij hawm, muab kev txhawb zog rau tom qab purification. Lub hollow fiber kem nrog qhov hnyav molecular txiav ntawm 300kd raug xaiv. Qhov pore loj no, thaum khaws ntau dua 85% ntawm tus kab mob, tuaj yeem tshem tawm ntau dua 90% ntawm cov protein tsis huv. Lub cheeb tsam ntawm daim nyias nyias yog 1600 cm², thiab qhov kev tawm mus yog 2-5 mL / cm², ua kom cov dej lim dej ua kom tiav hauv lub sijhawm zoo (hauv 2 teev). Cov ncauj lus kom ntxaws ultrafiltration concentration thiab purification cov ntaub ntawv muaj nyob rau hauv Table 1.
Table 1. Cov txiaj ntsig ntawm kev kuaj pom tag nrho ntawm ko taw- thiab - qhov ncauj kab mob antigens (hom O thiab A)
| Qauv Lub Npe | Kua Pub | Ntim (mL) | 146S Cov ntsiab lus (ug / ml) | 146S tus nqi rov qab (%) | Protein Concentration (ug / ml) | Protein Tshem Tawm Tus Nqi (%) |
|---|---|---|---|---|---|---|
| O- hom | Kev daws qhov qub | 5600 | 2.53 | 87.94 | 5040 | 94.38 |
| O- hom | Nco ntsoov | 500 | 24.92 | 87.94 | 3170 | 94.38 |
| Ib - hom | Kev daws qhov qub | 5600 | 3.37 | 94.80 | 5980 | 93.55 |
| Ib - hom | Nco ntsoov | 500 | 35.78 | 94.80 | 4318 | 93.55 |
Purification Method Kev txiav txim siab
Tam sim no, muaj ntau txoj hauv kev ua kom huv huv, tab sis txawm tias muaj tib txoj kev rau tib lub antigen, cov nyhuv purification feem ntau tsis txaus siab rau ntau yam, suav nrog cov cim antigen thiab cov txheej txheem tsis zoo. Piv txwv li, kev hloov pauv hauv cov qauv tsim qauv molecular lossis cov kab lis kev cai fermentation tuaj yeem hloov pauv cov qauv kab mob molecular, ua rau cov txiaj ntsig tsis zoo. Txoj kev qhia meej tsis tsim nyog tuaj yeem nce cov nqi raw khoom lossis tus nqi ntawm cov protein heterologous, ua rau lub siab ntawm kev ua kom huv hauv qab. Tsis tsim nyog ultrafiltration concentration, xws li sib txawv ultrafiltration membrane pore qhov ntau thiab tsawg, concentration thiab elution yam, inlet flow rates, pom tus nqi, thiab TMP, yuav ua rau tsawg antigen rov qab tus nqi. Kev xyaum dav dav qhia tau hais tias kev lim dej tsis zoo tuaj yeem ua rau muaj kev cuam tshuam loj heev hauv cov tsiaj tom qab txhaj tshuaj, uas ua rau tuag taus.
Xaus:Txoj kev tshawb no qhia tau hais tias, nyob rau hauv cov txheej txheem tangential ntws, los ntawm kev xaiv qhov tsim nyog daim nyias nyias pore loj (300kd), ntxuav piv, inlet flow rate, thiab TMP, ob qho tib si siab rov qab tus nqi thiab qhov siab tshaj plaws impurity tshem tawm tuaj yeem ua kom ntseeg tau.