Daim ntawv thov ntawm Ultrafiltration nyob rau hauv cov txheej txheem ntau lawm ntawm DTP tshuaj tiv thaiv

Daim ntawv thov ntawmUltrafiltration nyob rau hauv cov ntaub ntawvProductionProcess ntawmDTP Vaccine

 

1. Tshuaj tiv thaiv DTP

Pertussis, feem ntau hu ua hnoos hawb pob, yog tus kab mob ua pa nyuaj uas tshwm sim los ntawm Bordetella pertussis, feem ntau cuam tshuam rau cov menyuam mos thiab menyuam yaus. Nws kis tau los ntawm cov pa ua pa thiab muaj lub sijhawm incubation ntawm 7 mus rau 10 hnub. Tus neeg mob yuav tsim cov tsos mob ntawm hnoos qeev, nrog rau hnoos. Diphtheria yog ib qho kab mob ua pa nyuaj uas tshwm sim los ntawm Corynebacterium diphtheriae. Cov kab mob nkag mus rau hauv lub qhov ntswg mucosa lossis caj pas, proliferate, tsim exotoxins thiab ua rau lub zos o thiab cov nqaij mos necrosis. Tetanus, tshwm sim los ntawm Clostridium tetani, nkag mus rau tib neeg lub cev los ntawm daim tawv nqaij los yog mucous membrane qhov txhab. Nws hlob zoo nyob rau hauv cov xwm txheej anaerobic thiab tsim cov co toxins, ua rau spasmodic nqaij contractions.

Cov tshuaj tiv thaiv DTP yog ib qho tshuaj tiv thaiv ua ke uas muaj cov khoom siv zoo ntawm Bordetella pertussis, diphtheria toxin, thiab tetanus toxin sib xyaw rau hauv qee qhov kev faib ua feem thiab adsorbed rau ntawm aluminium hydroxide adjuvant. Nws yog siv los tiv thaiv peb kab mob: pertussis, diphtheria thiab tetanus.

 

2. Kev tsim cov tshuaj tiv thaiv DTP

Cov tshuaj tiv thaiv DTP feem ntau yog ib qho tshuaj tiv thaiv tsis muaj zog. Kev npaj tshuaj tiv thaiv DTP ua raws li cov txheej txheem tsim tshuaj tiv thaiv tsis muaj zog, feem ntau yog vim li cas kev nyab xeeb. Tam sim no, cov txheej txheem tsim cov tshuaj tiv thaiv DTP hauv Suav teb feem ntau ua raws li cov thev naus laus zis hauv xyoo 1970 thiab 1980s.Txawm li cas los xij, thoob plaws hauv cov txheej txheem, cov txheej txheem lim ntshav koom nrog ntau theem pom pom, cuam tshuam rau tag nrho cov txheej txheem ntawm cov khoom. Tangential flow membrane filtration khoom nrog qhib raws muaj qhov zoo nyob rau hauv downstream sib cais ntawm cov khoom lom, xws li qis shear rog thiab siab rov qab. Lawv tshwj xeeb tshaj yog haum rau kev sib cais ntawm cov qauv nrog cov concentrations siab, siab viscosities, thiab cov ntsiab lus siab, ua rau lawv tsim nyog rau cov txheej txheem ntawm cov tshuaj tiv thaiv DTP.

3. Daim ntawv thov ntawm Hollow Fiber Ultrafiltration Technology nyob rau hauv cov txheej txheem purification ntawm Cell-Dawb DTP Combined Vaccine Antigen Liquid

(1) Cov khoom siv thiab cov txheej txheem

①Sample

Decontaminated thiab refined cell-dawb DTP antigen tov (tom qab no hu ua decontaminated antigen tov).

②Main Reagents thiab Instruments

Txhaj-qib dej; sodium chloride (qib kws tshuaj); lub hnab lim ntshav; 10kDa hollow fiber ntau; peristaltic twj tso kua mis.

③ Ultrafiltration thiab Purification ntawm Cell-Free DTP Antigen Liquid

Ua ntej ntawm tag nrho cov, mloog zoo rau cov decontaminated antigen tov rau txog 5-fold thawj ntim siv 10kDa hollow fiber. Tom qab ntawd, pib cov txheej txheem lim thiab ntxuav. Tom ntej no, nres filtration thaum lub filtrate ntim nce mus txog kwv yees li 4 mus rau 5 npaug ntawm thawj ntim. Thaum kawg, yaug tag nrho cov kab ke thiab rov ua kom cov lim dej siv 1-fold ntim ntawm physiological saline.

④ Dialysis thiab Purification ntawm Cell-Free DTP Antigen Liquid

Aseptically ntim cov tshuaj decontaminated antigen rau hauv lub hnab lim ntshav thiab muab tso rau hauv 0.85% sterile sodium chloride tov thiab tom qab ntawd ua lim dej ntawm 2-8 degree .

⑤ Kev txheeb xyuas cov kua dej ntshiab huv

Raws li cov ntaub ntawv rov qab ultrafiltration thiab dialysis-recovered kua kuaj mus rau lub pre-purification ntim. Siv cov tshuaj decontaminated antigen los tswj thiab txheeb xyuas qhov purity ntawm cov qauv siv 12% SDS-PAGE. Ntsuas cov ntsiab lus hauv cov kev daws teeb meem thiab suav cov nqi rov qab tom qab centrifuging cov qauv ntawm 10, 000 rpm rau 15 feeb.

Cov txiaj ntsig

①Hollow Fiber Ultrafiltration Concentration thiab Ntxuav

Thaum lub sij hawm txheej txheem ultrafiltration concentration, 4,800 mL filtrate yog tau nyob rau hauv 70 feeb, ua rau ib tug 5- quav concentration. Qhov pom tus nqi tsis tu ncua, nrog qhov nruab nrab pom tus nqi ntawm 42 LMH (69.6 mL / min). Thaum lub sij hawm ntxuav, 5,300 mL filtrate yog tau nyob rau hauv 90 feeb, ua rau ib tug 4.4-fold ntxuav, thiab pom tus nqi tsis tu ncua, nrog qhov nruab nrab pom tus nqi ntawm 36 LMH (59.5 mL / min). Thaum kawg, yaug tag nrho cov kab ke nrog 1,200 mL physiological saline, thiab sib tov cov tshuaj yaug nrog cov tshuaj ntxuav yav dhau los (1,200 mL). Qhov kawg concentration ntawm cov kua yog 2.5-fold. Piv nrog rau ultrafiltration, dialysis yuav tsum tau hloov lub dialysate tsib zaug thiab yuav siv li ntawm 6 hnub.

② Kev txheeb xyuas cov kua dej ntshiab huv

SDS-PAGE kev tshuaj xyuas tau qhia tias qhov purity ntawm cell-free DTP tshuaj tiv thaiv antigen kua purified los ntawm ob qho tib si ultrafiltration thiab dialysis txoj kev tseem tsis tau hloov, raws li qhia nyob rau hauv daim duab hauv qab no. Tus nqi rov qab ntawm ultrafiltration purification yog me ntsis qis dua li ntawm dialysis purification, raws li qhia hauv cov lus hauv qab no.

Qauv	Cov ntsiab lus muaj zog (%)	Tus nqi rov qab (%)
Decontaminated Antigen Solution	0.872	/
Ultrafiltration-rov qab ua kua (txo)	0.803	92.1
Dialysis-recovered Liquid (txo)	0.813	93.2

 

Xaus

Hollow fiber ntau ultrafiltration yog cov tshuab sib cais membrane dav siv hauv kev lag luam xws li khoom noj khoom haus, fermentation, tshuaj, kev tiv thaiv ib puag ncig, thiab tshuaj lom neeg engineering rau kev sib cais, concentration, thiab purification ntawm cov khoom loj molecular. Hauv qhov kev sim no, hollow fiber ultrafiltration thev naus laus zis tau ua haujlwm los ntxuav cov kab mob cell-dawb DTP antigen daws. Tus kws sim piv nrog cov tshuaj lim ntshav hauv cov ntsiab lus ntawm kev ua kom huv huv thiab kev rov qab los. Cov txiaj ntsig tau pom tias ultrafiltration muaj kev ua haujlwm zoo dua hauv kev tshem tawm cov tshuaj detoxifying thiab cov pigments (ua tiav hauv 3 teev, piv rau 5-7 hnub rau kev lim ntshav) thiab muaj tus nqi rov zoo dua. Cov no qhia tias daim ntawv thov ntawm hollow fiber rau ultrafiltration purification ntawm cell-dawb DTP tshuaj tiv thaiv antigen kua yog ua tau. Nws tuaj yeem hloov cov txheej txheem lim dej uas twb muaj lawm.