Protein Solution Viscosity Control in Ultrafiltration

Ijikwa viscosity nke ngwọta protein dị oke mkpa maka imeziwanye usoro ntinye ultrafiltration na mmepụta biopharmaceutical. viscosity dị elu na ngwọta protein - karịsịa na nnukwu protein - na-emetụta arụmọrụ membrane, arụmọrụ usoro, na akụnụba na ngwa ntinye protein ultrafiltration ozugbo. viscosity ngwọta na-arị elu na ọdịnaya protein n'ihi nchịkọta mgbochi ahụ na mmekọrịta electrostatic, nke na-eme ka iguzogide mmiri na mbelata nrụgide na-agafe na membrane ultrafiltration dịkwuo elu. Nke a na-eme ka obere fluxes intravement na oge ọrụ dị ogologo, karịsịa na usoro nzacha transverse flow (TFF).

Nrụgide Transmembrane (TMP), ike na-akpali ultrafiltration, nwere njikọ chiri anya na viscosity. Ịrụ ọrụ n'èzí oke nrụgide transmembrane nkịtị na-eme ka mmebi akpụkpọ ahụ dịkwuo ngwa ngwa ma na-eme ka polarization nke mkpokọta dịkwuo njọ—mkpokọta protein dị nso na akpụkpọ ahụ nke na-eme ka viscosity mpaghara dịkwuo elu mgbe niile. Ma polarization nke mkpokọta na ntụpọ akpụkpọ ahụ na-eme ka arụmọrụ akpụkpọ ahụ ultrafiltration belata ma nwee ike ime ka ndụ akpụkpọ ahụ dị mkpụmkpụ ma ọ bụrụ na emeghị ya. Ọrụ nnwale na-egosi na ntụpọ akpụkpọ ahụ na polarization nke mkpokọta na ultrafiltration na-apụta ìhè karịa na ụkpụrụ TMP dị elu yana nri viscous karịa, na-eme ka njikwa TMP oge dị mkpa iji bulie mmepụta na ibelata ugboro nhicha.

Ịkwalite njupụta ultrafiltration chọrọ atụmatụ agbakwunyere:

Nnwale viscosity nke ngwọta proteinNyocha viscosity nkịtị—ijindị na-enyocha ihe ndị dị n'ahịrị—nyere aka ịkọ amụma ọnụego nzacha na ịtụ anya nsogbu usoro, na-akwado mgbanwe usoro ngwa ngwa.
Nhazi nri: Ịgbanwe pH, ike ionic, na okpomọkụ nwere ike ibelata viscosity ma belata friability. Dịka ọmụmaatụ, itinye sodium ion na-eme ka mmiri ghara ịdị n'etiti protein, na-ebelata mkpokọta na friability, ebe calcium ion na-akwalite njikọ protein na friability.
Ojiji nke ihe mgbakwunye: Itinye ihe ndị na-ebelata viscosity n'ime ngwọta protein dị elu na-eme ka akpụkpọ ahụ nwee ike ịbanye nke ọma ma na-ebelata nrụgide transmembrane na ultrafiltration, na-eme ka arụmọrụ zuru oke dịkwuo mma.
Usoro mmiri dị elu: Ịbawanye ọsọ nke ikuku na-agafe agafe, iji mgbanwe nke ikuku na-agafe agafe, ma ọ bụ iji ntụtụ ikuku na-emebi akwa mmebi. Usoro ndị a na-enyere aka ịnọgide na-enwe mmiri na-abanye ma belata ugboro ole a na-agbanwe agbanwe nke akpụkpọ ahụ site na ibelata nhazi nke ihe nkwụnye.
Nhọrọ na nhicha akpụkpọ ahụỊhọrọ akpụkpọ ahụ ndị na-eguzogide kemịkalụ (dịka ọmụmaatụ, SiC ma ọ bụ ngwakọta thermosalient) na ime ka ugboro ole a na-ehicha akpụkpọ ahụ dị mma (dịka ọmụmaatụ, nhicha sodium hypochlorite) dị oke mkpa maka ịgbatị ndụ akpụkpọ ahụ ma belata ọnụ ahịa ọrụ.

N'ozuzu, njikwa viscosity dị irè na njikwa TMP bụ isi ihe na-eme ka arụmọrụ nke usoro ntinye ultrafiltration na-aga nke ọma, na-emetụta mmepụta ngwaahịa ozugbo, ugboro ole nhicha akpụkpọ ahụ, na ogologo oge nke akụ akpụkpọ ahụ dị oke ọnụ.

Ịghọta Ọdịiche Dị n'etiti Ngwọta Protein na Ultrafiltration

1.1. Kedu ihe bụ Viscosity nke Ngwọta Protein?

Viscosity na-akọwa otú mmiri si eguzogide mmiri; na ngwọta protein, ọ na-egosi etu esemokwu molekul si egbochi mmegharị. Ngalaba SI maka viscosity bụ Pascal-second (Pa·s), mana a na-ejikarị centipoise (cP) eme ihe maka mmiri bayọlọji. Viscosity na-emetụta kpọmkwem otu esi agba mmiri ma ọ bụ nzacha protein ngwa ngwa n'oge mmepụta ma na-emetụta nnyefe ọgwụ, karịsịa maka biotherapeutics nwere nnukwu njupụta.

Njupụta protein bụ ihe kacha emetụta viscosity. Ka ọkwa protein na-arị elu, mmekọrịta dị n'etiti molekul na mkpokọta na-abawanye, na-eme ka viscosity rịgoro, ọtụtụ mgbe na-abụghị nke kwụ ọtọ. N'elu oke ụfọdụ, mmekọrịta protein na protein na-egbochi mgbasa n'ime ngwọta ahụ. Dịka ọmụmaatụ, ngwọta monoclonal antibody a na-ejikarị eme ihe na ọgwụ na-erukarị ọkwa viscosity nke na-ama aka n'ịgba ọgwụ ma ọ bụ na-egbochi ọnụego nhazi.

Ụdị ndị na-ebu amụma banyere viscosity na ngwọta protein a gbakọtara agwakọta ugbu a na-etinye usoro geometry molekul na njikọta. Ọdịdị protein—ma ọ dị ogologo, gburugburu ụwa, ma ọ bụ nke na-adịkarị mfe ịnakọta—na-emetụta viscosity nke ukwuu na oke njupụta. Mmepe ọhụrụ na nyocha microfluidic na-eme ka nha viscosity ziri ezi site na obere olu ihe nlele, na-eme ka nyocha ngwa ngwa nke usoro protein ọhụrụ dị mfe.

1.2. Otu Viscosity si agbanwe n'oge Ultrafiltration

N'oge ultrafiltration, polarization nke njupụta na-achịkọta protein ngwa ngwa na njikọ membrane-solution. Nke a na-emepụta oke njupụta mpaghara ma na-eme ka viscosity dịkwuo nso na akpụkpọ ahụ. Oke viscosity na mpaghara a na-egbochi mbufe oke ma na-ebelata mmiri ịbanye n'ime.

Mbelata nke mkpokọta ihe dị iche na mmetọ akpụkpọ ahụ. Mbelata nke mkpokọta ihe dị iche na mmetọ akpụkpọ ahụ na-agbanwe agbanwe, na-eme n'ime nkeji ole na ole ka nzacha na-aga n'ihu. Ma e jiri ya tụnyere ya, mmetọ ahụ na-etolite ka oge na-aga ma na-agụnyekarị ntinye ma ọ bụ mgbanwe kemịkalụ na-enweghị mgbagha na elu akpụkpọ ahụ. Nchọpụta ziri ezi na-enye ohere ka a na-enyocha oyi akwa polarization nke mkpokọta ihe dị iche iche n'oge, na-ekpughe mmetụta ya na ọsọ gafere-flow na nrụgide transmembrane. Dịka ọmụmaatụ, ịbawanye ọsọ ma ọ bụ mbelata nrụgide transmembrane (TMP) na-enyere aka imebi oyi akwa ókè viscous, na-eweghachi flux.

Usoro ọrụ na-emetụta omume viscosity ozugbo:

Nrụgide Transmembrane (TMP): TMP dị elu na-eme ka polarization dịkwuo elu, na-eme ka viscosity mpaghara dịkwuo elu ma na-ebelata flux.
Ọsọ mbufe-agafe: Ọsọ ọsọ na-ebelata mkpokọta, na-eme ka ọ dị mfe ịnweta oke ọkụ n'akụkụ akpụkpọ ahụ.
Ugboro nhicha akpụkpọ ahụ: Ịsacha ihe mgbe niile na-ebelata mkpokọta ogologo oge ma na-ebelata mfu arụmọrụ nke na-akpata viscosity.

Usoro ntinye nke Ultrafiltration ga-eme ka paramita ndị a dịkwuo mma iji belata mmetụta viscosity na-adịghị mma ma nọgide na-enwe mmepụta.

1.3. Njirimara Ngwọta Protein Na-emetụta Viscosity

Ibu molekụlananhazina-ekpebikarị viscosity. Protein ma ọ bụ ngwakọta buru ibu ma dị mgbagwoju anya na-enye viscosity dị elu n'ihi mgbochi mmegharị na ike dị ukwuu nke intermolecular. Ọdịdị nke protein na-agbanwekwa usoro mmiri - agbụ ndị toro ogologo ma ọ bụ ndị na-enwe ike ịchịkọta na-akpata nguzogide karịa protein globular dị obere.

pHỌ na-emetụta oke ibu na ike ịgbaze protein. Ịgbanwe pH ngwọta dị nso na ebe isoelectric nke protein na-ebelata ibu net, na-ebelata mwepu protein-protein, ma na-ebelata viscosity nwa oge, na-eme ka nzacha dị mfe. Dịka ọmụmaatụ, ọrụ ultrafiltration dị nso na ebe isoelectric nke BSA ma ọ bụ IgG nwere ike ime ka ntinye mmiri na nhọrọ nkewa dịkwuo mma.

Ike ionicna-emetụta viscosity site na ịgbanwe oyi akwa okpukpu abụọ nke eletriki gburugburu protein. Ike ionic mụbara na-enyocha mmekọrịta electrostatic, na-akwalite nnyefe protein site na membranes mana ọ na-emekwa ka ihe egwu nke nchịkọta na mmụba viscosity kwekọrọ. Mmekọrịta dị n'etiti arụmọrụ nnyefe na nhọrọ na-adaberekarị na nhazi nnu dị mma na nhazi buffer.

Enwere ike iji obere ihe mgbakwunye molekul—dịka arginine hydrochloride ma ọ bụ guanidine—belata viscosity. Ihe ndị a na-emebi ihe ndị na-adọta hydrophobic ma ọ bụ electrostatic, belata mkpokọta, ma melite njirimara mmiri mmiri ngwọta. Okpomọkụ na-arụ ọrụ dị ka ihe na-agbanwe agbanwe ọzọ; obere okpomọkụ na-eme ka viscosity dịkwuo elu, ebe okpomọkụ ọzọ na-ebelatakarị ya.

Ihe dị mkpa maka nha viscosity nke protein ngwọta bụ:

Nkesa ibu molekụla
Ngwakọta nke ngwọta (nnu, ihe mgbakwunye, ihe mgbakwunye)
Nhọrọ nke sistemụ pH na nchekwa
Ntọala ike Ionic

Ihe ndị a dị oke mkpa maka ime ka arụmọrụ akpụkpọ ahụ ultrafiltration dịkwuo mma ma hụ na ọ na-adịgide n'otu oge n'usoro ntinye uche na usoro TFF.

Isi ihe dị mkpa maka ntinye protein Ultrafiltration

Ụkpụrụ nke Oge Ntinye Uche Ultrafiltration

Njupụta protein Ultrafiltration na-arụ ọrụ site n'itinye nrụgide transmembrane (TMP) n'ofe akpụkpọ ahụ nke nwere ike ịbanye, na-akpali ihe mgbaze na obere ihe mgbaze site na ya ebe ọ na-ejide protein na molekul buru ibu. Usoro a na-eji nhọrọ permeation dabere na nha molekul, yana mbelata ibu molekul nke membrane (MWCO) na-akọwa oke nha molekul ndị na-agafe. Protein ndị gafere MWCO na-agbakọta n'akụkụ retentate, na-eme ka ntinye ha dịkwuo elu ka a na-ewepụ permeability.

Usoro ntinye ultrafiltration na-elekwasị anya na mbelata olu na ịba ụba nke ngwọta protein. Ka nzacha na-aga n'ihu, viscosity nke ngwọta protein na-ebilikarị, na-emetụta mmụba na ihe achọrọ TMP. Protein echekwara nwere ike ịkpakọrịta na ibe ha na akpụkpọ ahụ, na-eme ka usoro ụwa dị adị dịkwuo mgbagwoju anya karịa mwepụ nha dị mfe. Mmekọrịta electrostatic, nchịkọta protein, na njirimara ngwọta dịka pH na ike ionic na-emetụta nsonaazụ njide na nkewa. N'ọnọdụ ụfọdụ, njem advective na-achịkwa mgbasa, karịsịa na akpụkpọ ahụ nwere nnukwu pores, na-eme ka atụmanya dabere naanị na nhọrọ MWCO [lee nchịkọta nyocha].

Nkọwa nke Nzacha Ọsọ Transverse (TFF)

Nzacha mmiri transverse, nke a na-akpọkwa tangential flow filter (TFF), na-ebuga mmiri protein ahụ n'elu akpụkpọ ahụ site na tangential. Ụzọ a dị iche na nzacha dead-end, ebe mmiri na-adaba na akpụkpọ ahụ, na-akwanye ihe ndị ahụ ozugbo na nzacha ahụ.

Ihe dị iche na mmetụta dị iche iche:

Njikwa Mmebi:TFF na-ebelata mkpokọta protein na ihe ndị dị n'ime ya, nke a maara dị ka nhazi achịcha, site n'ịnapụ ihe ndị nwere ike imebi akpụkpọ ahụ mgbe niile. Nke a na-eme ka mmiri na-agbagharị nke ọma ma na-adị mfe idozi.
Nchekwa protein:TFF na-akwado njikwa ka mma nke polarization nke mkpokọta ihe—ogwe nke molekul ndị a na-ejide n'akụkụ akpụkpọ ahụ—nke, ọ bụrụ na achịkwaghị ya, nwere ike ibelata nhọrọ nkewa ma mee ka ntụpọ dịkwuo mma. Usoro mgbanwe dị na TFF na-ebelata mmetụta a, na-enyere aka ịnọgide na-enwe oke njigide protein na arụmọrụ nkewa.
Ịkwụsi ike na mgbanwe:TFF na-eme ka oge ọrụ dị ogologo na mgbanwe na-agbanwe agbanwe, na-eme ka arụmọrụ dịkwuo mma na usoro nwere nnukwu protein ma ọ bụ nri bara ụba na ihe ndị dị ndụ. N'aka nke ọzọ, nzacha na-adịghị mma na-egbochi ngwa ngwa site na mmebi, ibelata mmepụta na-ebelata ma chọọ nhicha ugboro ugboro.

Ụdị TFF dị elu, dịka mgbanwe nke mmiri ọkụ (ATF), na-akpaghasị mmebi na nhazi achịcha site na ịtụgharị ma ọ bụ ịgbanwe ọsọ ọkụ ọkụ mgbe niile, na-agbatị ndụ nzacha na imeziwanye mmepụta protein [lee nchịkọta nyocha]. Na ntọala TFF ochie na nke dị elu, ntọala ọrụ - dị ka TMP, ọsọ ọsọ ọkụ ọkụ, na ugboro nhicha - ga-abụrịrị nke ahaziri maka sistemụ protein kpọmkwem, ụdị akpụkpọ ahụ, na ntinye uche iji mee ka arụmọrụ dịkwuo mma ma belata mmebi.

Nrụgide Transmembrane (TMP) na Ultrafiltration

3.1. Gịnị bụ nrụgide Transmembrane?

Nrụgide Transmembrane (TMP) bụ ọdịiche nrụgide n'ofe akpụkpọ nzacha, na-ebuga ihe mgbaze site n'akụkụ nri gaa n'akụkụ permerate. TMP bụ isi ike dị n'azụ usoro nkewa na ultrafiltration, na-enye ohere ka ihe mgbaze gafere na akpụkpọ ahụ ma na-ejide protein na macromolecules ndị ọzọ.

Usoro TMP:

Ihe dị mfe dị iche: TMP = P_feed − P_permeate
Usoro injinia: TMP = [(P_feed + P_retentate)/2] − P_permeate
N'ebe a, P_feed bụ nrụgide ntinye, P_retentate bụ nrụgide ọpụpụ n'akụkụ retentate, P_permeate bụkwa nrụgide akụkụ intermete. Itinye nrụgide retentate (ma ọ bụ concentrate) na-enye uru ziri ezi karịa n'elu akpụkpọ ahụ, na-aza ajụjụ maka mgbanwe nrụgide nke iguzogide mmiri na ntụpọ na-akpata.
Nri nrụgide na ọnụego eruba
Nrụgide na-adịgide adịgide (mgbe ọ dị mkpa)
Nrụgide permeate (ọtụtụ mgbe ikuku)
Mgbochi akpụkpọ ahụ
TMP na-agbanwe dabere na ụdị akpụkpọ ahụ, nhazi sistemụ, na ọnọdụ usoro.

Njikwa Mgbanwe:

3.2. TMP na Usoro Ultrafiltration

TMP na-arụ ọrụ dị mkpa na ntinye protein ultrafiltration, na-akwalite ngwọta protein site na akpụkpọ ahụ. Nrụgide ahụ ga-adị oke elu iji merie iguzogide site na akpụkpọ ahụ na ihe ọ bụla a chịkọtara mana ọ gaghị adị elu nke na ọ na-eme ka mmebi ahụ dịkwuo ngwa ngwa.

Mmetụta nke Viscosity Ngwọta na Ntinye Protein

Viscosity nke ngwọta protein:Ọbara ọbara dị elu na-eme ka iguzogide mmiri dịkwuo elu, na-achọ ka TMP dị elu iji nọgide na-enwe otu mmiri ọbara dị n'ime. Dịka ọmụmaatụ, itinye glycerol na nri ma ọ bụ ịrụ ọrụ na protein ndị a chịkọtara na-eme ka mmiri ọbara dịkwuo elu, ya mere TMP na-arụ ọrụ dị mkpa.
Ọnụọgụgụ protein:Ka njupụta na-abawanye n'oge usoro ntinye ultrafiltration, viscosity ngwọta na-arị elu, TMP na-abawanye, ihe egwu nke mmebi akpụkpọ ahụ ma ọ bụ polarization nke njupụta na-amụba.
Iwu Darcy:TMP, mmiri mmiri na-asọpụta (J), na viscosity (μ) jikọtara site na TMP = J × μ × R_m (nguzogide akpụkpọ ahụ). Maka ngwọta protein dị elu, nhazi TMP dị mkpa maka nzacha ultrafiltration dị irè.

Ihe atụ:

Nzacha nke ngwọta antibody dị oke mkpa chọrọ njikwa TMP nke ọma iji lụso mmụba nke viscosity ọgụ.
PEGylation ma ọ bụ mgbanwe protein ndị ọzọ na-agbanwe mmekọrịta na akpụkpọ ahụ, na-emetụta TMP achọrọ maka mmụba achọrọ.

3.3. Nlekota na Imeziwanye TMP

Ịdebe TMP n'ime ime ụlọoke nrụgide transmembrane nkịtịdị oke mkpa maka arụmọrụ akpụkpọ ahụ ultrafiltration kwụsiri ike na ịdị mma nke ngwaahịa. Ka oge na-aga, ka ultrafiltration na-aga n'ihu, polarization nke nguzosi ike na ntụpọ nwere ike ime ka TMP rịa elu, mgbe ụfọdụ ngwa ngwa.

Omume Nleba Anya:

Nlekota oge n'ezie:A na-enyocha TMP site na ntinye, retentate, na intermetendị na-ebugharị nrụgide.
Raman Spectroscopy:A na-eji ya maka nlekota protein na ihe ndị na-enyere aka na-anaghị emerụ ahụ, na-eme ka njikwa TMP na-agbanwe agbanwe n'oge ultrafiltration na diafiltration dị mfe.
Njikwa dị elu:Nzacha Kalman Extended (EKF) nwere ike ịhazi data sensọ, na-agbanwe TMP na akpaghị aka iji zere mmebi gabigara ókè.
Debe TMP mbụ n'ime oke nkịtị:Ọ dịghị oke ala iji belata mmiri na-asọfe, ọ dịghịkwa oke elu iji zere mmebi ngwa ngwa.
Gbanwee TMP ka viscosity na-abawanye:N'oge usoro ntinye uche nke ultrafiltration, bulie TMP nwayọ nwayọ dịka ọ dị mkpa.
Njikwa mmiri oriri na pH:Ịbawanye mmiri ọkụ ma ọ bụ ibelata TMP na-ebelata polarization nke ntinye uche na mmebi.
Nhicha na nnọchi akpụkpọ ahụ:A na-ejikọta TMP dị elu na nhicha ugboro ugboro na mbelata ndụ akpụkpọ ahụ.

Atụmatụ kachasị mma:

Ihe atụ:

Mmebi nke nchara n'ime usoro nhazi protein na-eduga na mmụba nke TMP na mbelata mmiri na-agbapụta, nke chọrọ nhicha ma ọ bụ dochie akpụkpọ ahụ iji weghachi ọrụ nkịtị.
Ọgwụgwọ tupu enzyme (dịka ọmụmaatụ, itinye pectinase) nwere ike ibelata TMP ma gbasaa ndụ akpụkpọ ahụ n'oge ultrafiltration nke protein rapeseed dị elu.

3.4. TMP na Sistemụ TFF

Nzacha mmiri nke Tangential (transverse) (TFF) na-arụ ọrụ site n'itinye mmiri nri n'ofe akpụkpọ ahụ kama site na ya kpọmkwem, na-emetụta ike TMP nke ukwuu.

Iwu na Nhazi nke TMP

Nrụgide transmembrane TFF (TFF TMP):A na-ejikwa ya site n'ịchịkwa ma ọnụego mmiri nri na nrụgide mgbapụta iji zere oke TMP ebe ọ na-eme ka mmiri mmiri na-asọfe nke ọma.
Nhazi paramita:Ịba ụba nke mmiri nri na-ebelata ntinye protein n'ógbè ahụ, na-eme ka TMP kwụsie ike, ma na-ebelata mmebi akpụkpọ ahụ.
Nhazi kọmputa:Ụdị CFD na-ebu amụma ma na-eme ka TFF TMP dị mma maka mgbake ngwaahịa kachasị, ịdị ọcha, na mmepụta - karịsịa dị oke mkpa maka usoro dịka mRNA ma ọ bụ mwepụ nke vesicle extracellular.

Ihe atụ:

N'ime bioprocessing, TMP TFF kacha mma na-enye mgbake mRNA karịa 70% na-enweghị mmebi, na-arụ ọrụ karịa usoro ultracentrifugation.
Njikwa TMP na-agbanwe agbanwe, nke ihe nlereanya mgbakọ na mwepụ na nzaghachi sensọ na-enye, na-ebelata ugboro mgbanwe akpụkpọ ahụ ma na-eme ka ndụ akpụkpọ ahụ ka mma site na ibelata ntụpọ.

Isi ihe dị mkpa:

A ghaghị ijikwa nrụgide transmembrane TMP nke ọma na TFF iji nọgide na-arụ ọrụ nke ọma, na-agbanwe agbanwe, na ahụike akpụkpọ ahụ.
Nhazi TMP nke sistemụ na-ebelata ọnụ ahịa ọrụ, na-akwado mgbake ngwaahịa dị ọcha, ma na-agbatị ndụ akpụkpọ ahụ na ultrafiltration protein na usoro ndị metụtara ya.

Usoro Mmebi na Mmekọrịta Ha na Viscosity

Ụzọ Ndị Isi Na-emebi Ihe na Ultrafiltration Protein

Mmepụta protein ultrafiltration na-adabere n'ọtụtụ ụzọ dị iche iche esi arụ ọrụ:

Mmebi nke ire ere:Ọ na-eme mgbe ngwaahịa nchara—karịsịa iron oxides—na-agbakọta n'elu akpụkpọ ahụ. Ihe ndị a na-ebelata mmiri na-agbapụta ma na-esi ike iwepụ ya site na iji ihe nhicha kemịkalụ nkịtị. Nchacha nchara na-eduga na mfu na arụmọrụ akpụkpọ ahụ na-adịgide adịgide ma na-eme ka ugboro ugboro nnọchi akpụkpọ ahụ dịkwuo elu ka oge na-aga. Mmetụta ya dị oke njọ karịsịa na akpụkpọ ahụ PVDF na PES ejiri mee ihe na ọgwụgwọ mmiri na itinye protein.

Ndozi ihe dị ndụ:Ihe ndị a na-ebutekarị site na protein dịka bovine serum albumin (BSA), ma nwee ike ime ka ọ sie ike ma ọ bụrụ na e nwere ihe ndị ọzọ dị ndụ dịka polysaccharides (dịka ọmụmaatụ, sodium alginate). Usoro ndị a gụnyere ịmịnye ihe n'ime akpụkpọ ahụ, itinye oghere n'ime oghere ahụ, na ịmepụta oyi akwa achịcha. Mmetụta ndị a na-eme mgbe ọtụtụ ihe ndị dị ndụ dị, yana sistemụ ndị na-eme ka ihe dị nro na-enwe mmerụ ahụ dị njọ karịa nri protein otu.

Oke nke Ịhazi Uche:Ka ultrafiltration na-aga n'ihu, protein ndị e jigide na-agbakọta n'akụkụ elu akpụkpọ ahụ, na-eme ka mkpokọta na viscosity dịkwuo elu. Nke a na-emepụta oyi akwa polarization nke na-eme ka ohere mmebi dịkwuo elu ma na-ebelata flux. Usoro a na-eme ngwa ngwa ka usoro ntinye ultrafiltration na-aga n'ihu, nke nrụgide transmembrane na dynamics na-emetụta kpọmkwem.

Mbelata Colloidal na Mgbakwunye Foulant:Ihe colloidal (dịka ọmụmaatụ, silica, mineral inorganic) nwere ike imekọrịta ihe na protein, na-emepụta akwa mkpokọta dị mgbagwoju anya nke na-eme ka mmebi akpụkpọ ahụ ka njọ. Dịka ọmụmaatụ, ọnụnọ nke colloidal silica na-ebelata ọnụego mmiri, karịsịa mgbe ejikọtara ya na ihe organic ma ọ bụ n'okpuru ọnọdụ pH na-adịghị mma.

Mmetụta nke Viscosity Ngwọta na Mmepe Mbelata

Viscosis nke ngwọta protein na-emetụta nke ukwuu na mmepụta akpụkpọ ahụ mebiri emebi:

Mmebi Ngwa ngwa:Nnukwu oke mmiri protein na-eme ka iguzogide mbufe azụ nke ihe ndị na-agbaze agbaze dịkwuo elu, na-eme ka nhazi oyi akwa achịcha dị ngwa ngwa. Nke a na-eme ka nrụgide transmembrane (TMP) dịkwuo elu, na-eme ka akpụkpọ ahụ dị ngwa ngwa ma na-emebi ihe.

Mmetụta nke Ngwakọta Ngwọta:Ụdị protein na-agbanwe viscosity; protein globular (dịka ọmụmaatụ, BSA) na protein ndị gbatịrị agbatị na-akpa àgwà dị iche iche gbasara mmiri na polarization. Ịtinye ihe ndị dị ka polysaccharides ma ọ bụ glycerol na-eme ka viscosity dịkwuo elu nke ukwuu, na-akwalite ntụpọ. Ihe mgbakwunye na nchịkọta protein na oke njupụta na-eme ka ọsọ nke membranes na-emechi emechi dịkwuo elu, na-ebelata ma flux na ndụ membrane ozugbo.

Nsonaazụ Ọrụ:Ọbara dị elu chọrọ mmụba TMP iji kwado ọnụego nzacha na usoro nzacha transverse flow. Ogologo oge ikpughe na TMP dị elu na-eme ka mmebi a na-apụghị ịgbanwe agbanwe dịkwuo mma, na-emekarị ka ọ dị mkpa ka a na-ehicha akpụkpọ ahụ ugboro ugboro ma ọ bụ dochie akpụkpọ ahụ na mbụ.

Ọrụ nke Njirimara Nri

Àgwà nri—ya bụ, àgwà protein na kemistri mmiri—na-ekpebi oke mmebi ahụ:

Nha na Nkesa Protein:Protein buru ibu ma ọ bụ nke a chịkọtara nwere ike ime ka oghere na-amachi ma na-agbakọta achịcha, na-eme ka viscosity na mkpakọ dịkwuo elu n'oge ultrafiltration protein.

pH:Oke pH na-eme ka electrostatic repulsion dịkwuo elu, na-egbochi protein ịchịkọta nso membrane ahụ, si otú a na-ebelata mmetọ. N'ụzọ dị iche, ọnọdụ acidic na-ebelata mmịpụta, karịsịa maka colloidal silica, na-eme ka mmetọ akpụkpọ ahụ ka njọ ma na-ebelata ọnụego flux.

Okpomọkụ:Okpomọkụ usoro dị ala na-ebelata ike kinetic, nke nwere ike ibelata ọnụego mmebi ma meekwa ka mmiri ghara ịdị. Okpomọkụ dị elu na-eme ka mmebi ahụ dịkwuo ngwa ngwa mana ọ nwekwara ike ime ka nhicha dịkwuo mma.

Ihe Colloidal/Inorganic:Ọnụnọ nke silica colloidal ma ọ bụ ọla na-eme ka ntụpọ dịkwuo njọ, ọkachasị n'ọnọdụ acidic. Mkpụrụ silica na-eme ka viscosity ngwọta zuru oke dịkwuo elu ma na-egbochi oghere ahụ, na-eme ka ntinye ultrafiltration ghara ịrụ ọrụ nke ọma ma na-ebelata ndụ na arụmọrụ nke akpụkpọ ahụ dum.

Ngwakọta Ionic:Ịtinye ụfọdụ ụdị ionic (Na⁺, Zn²⁺, K⁺) nwere ike ibelata ntụpọ site na ịgbanwe ike electrostatic na hydration n'etiti protein na membranes. Agbanyeghị, ion dị ka Ca²⁺ na-akwalite nchịkọta ma na-eme ka ikike ntụpọ dịkwuo elu.

Ihe atụ:

N'oge nzacha mmiri transverse, nri bara ụba na protein dị arọ nke molekul na oke viscosity ga-enwe mbelata ọsọ ọsọ, na-abawanye n'ihi nhicha na usoro nnọchi.
Mgbe mmiri nri nwere silica colloidal ma mee ka ọ dị ka acid, nchịkọta na ntinye silica na-akawanye njọ, na-eme ka ọnụego mmebi dịkwuo elu ma na-ebelata arụmọrụ akpụkpọ ahụ.

Na nchịkọta, nghọta nke mmekọrịta dị n'etiti viscosity ngwọta, ụdị ntụpọ, na njirimara nri dị oke mkpa maka ime ka ntinye ultrafiltration dịkwuo mma, belata ntụpọ akpụkpọ ahụ, na ime ka ndụ akpụkpọ ahụ ka mma.

Nhazi nke Mkpuchi na Njikwa Ya

Gịnị bụ Polarisation nke Ịhazi Uche?

Mkpokọta mkpụrụ ndụ ihe nketa bụ mkpokọta mmiri e jigide—dị ka protein—n'ime membrane/solution interface n'oge ultrafiltration. N'ihe gbasara mmiri protein, ka mmiri mmiri na-asọba na akpụkpọ ahụ nke na-erugharị n'ime ọkara-permeable, protein ndị akpụkpọ ahụ jụrụ na-agbakọta n'ime obere oyi akwa dị nso n'elu. Mkpokọta a na-ebute oke njupụta: nnukwu njupụta protein kpọmkwem na akpụkpọ ahụ, dị ala karịa na ngwọta buru ibu. Ihe a na-agbanwe agbanwe ma na-achịkwa ya site na ike hydrodynamic. Ọ dị iche na ntụpọ akpụkpọ ahụ, nke gụnyere itinye ihe ma ọ bụ mmịpụta na-adịgide adịgide n'ime ma ọ bụ na akpụkpọ ahụ.

Otu Polarization nke Ntụnye Uche si eme ka Viscosity na Fouling ka njọ

N'elu akpụkpọ ahụ, nchịkọta protein na-aga n'ihu na-emepụta akwa ókè nke na-eme ka mkpokọta solute mpaghara dịkwuo elu. Nke a nwere mmetụta abụọ dị mkpa:

Mmụba nke Viscosity nke Mpaghara:Ka njupụta protein na-arị elu nso akpụkpọ ahụ, njupụta nke mmiri protein dị na mpaghara a na-abawanyekwa. Njupụta dị elu na-egbochi mbufe azụ nke solute pụọ na akpụkpọ ahụ, na-eme ka njupụta dịkwuo elu ma na-emepụta usoro nzaghachi nke iguzogide mmụba na mmiri. Nke a na-ebute mbelata mmiri intravelation na mkpa ike dị elu maka nzacha na-aga n'ihu.

Nkwadebe nke Mkpuchi Akpụkpọ Ahụ:Nnukwu protein dị nso na akpụkpọ ahụ na-eme ka ohere nke nchịkọta protein dịkwuo elu, na, n'ụfọdụ sistemụ, ịmepụta oyi akwa gel. Akwa a na-egbochi oghere akpụkpọ ahụ ma na-eme ka iguzogide mmiri dịkwuo elu. Ọnọdụ ndị dị otú ahụ chara acha maka mmalite nke mmebi a na-apụghị ịgbanwe agbanwe, ebe mkpokọta protein na ihe ndị na-adịghị ọcha na-ejikọ na matrix membrane ahụ n'ụzọ anụ ahụ ma ọ bụ kemịkalụ.

Foto nnwale (dịka ọmụmaatụ, electron microscopy) na-egosi nchịkọta ngwa ngwa nke mkpokọta protein nanos dị na membrane ahụ, nke nwere ike itolite ruo nnukwu nkwụnye ma ọ bụrụ na ejighị ntọala ọrụ nke ọma.

Atụmatụ iji belata oke ọdịiche dị n'etiti ihe ndị a na-etinye n'ime ihe.

Ijikwa polarization nke mkpokọta na ultrafiltration protein concentration ma ọ bụ transverse flow filtration chọrọ ụzọ abụọ: ịgbanwe hydrodynamics na ịgbanwe paramita ọrụ.

Nhazi ọsọ nke obe:
Ịbawanye ọsọ nke mmiri na-agafe agafe na-eme ka mmiri na-asọfe n'ofe akpụkpọ ahụ dịkwuo elu, na-akwalite shear ma na-eme ka oyi akwa ókè nke ihe dị n'ime ya dị nro. Ịcha ihe siri ike karị na-ekpochapụ protein ndị a chịkọtara n'elu akpụkpọ ahụ, na-ebelata polarization na ihe ize ndụ nke mmebi. Dịka ọmụmaatụ, iji ihe ngwakọta static ma ọ bụ itinye gas na-emebi oyi akwa solute, nke kachasị mma ime ka mmiri na-asọfe na arụmọrụ dị mma na usoro nzacha transverse flow.

Ịgbanwe Usoro Ọrụ:

Nrụgide Transmembrane (TMP):TMP bụ ọdịiche nrụgide dị n'ofe akpụkpọ ahụ na ike mkpali maka nzacha ultrafiltration. Agbanyeghị, ịkwanye TMP elu iji mee ka nzacha dị ngwa ngwa nwere ike imebi site na ime ka polarization nke njupụta dịkwuo elu. Ịgbaso oke nrụgide transmembrane nkịtị - anaghị agafe oke edobere maka nzacha protein - na-enyere aka igbochi oke nchịkọta solute na mmụba metụtara ya na viscosity mpaghara.

Ọnụego Ịkpụcha:Ọnụego shear, nke bụ ọrụ nke ọsọ na-agafe agafe na nhazi ọwa, na-arụ ọrụ dị mkpa na mgbanwe njem solute. Nnukwu shear na-eme ka oyi akwa polarization dị gịrịgịrị ma na-agagharị, na-enye ohere imeghari mpaghara solute-depleted dị nso na membrane ahụ ugboro ugboro. Ịba ụba ọnụego shear na-ebelata oge protein ga-agbakọta ma belata mmụba viscosity na interface ahụ.

Njirimara nri:Ịgbanwe àgwà nke ihe ngwọta protein na-abata—dịka ibelata viscosity nke ihe ngwọta protein, ibelata ọdịnaya mkpokọta, ma ọ bụ ijikwa pH na ike ionic—nwere ike inye aka belata oke na mmetụta nke polarization nke njupụta. Mgbanwe tupu ọgwụgwọ na nhazi nri nwere ike ime ka arụmọrụ membrane ultrafiltration dịkwuo mma ma gbasaa ndụ akpụkpọ ahụ site na ibelata ugboro ole a na-ehicha akpụkpọ ahụ.

Ihe Nlereanya Ngwa:
Ụlọ ọrụ na-eji nzacha mmiri tangential (TFF) iji mee ka ọgwụ mgbochi monoclonal dịgide na-eji ọsọ mmiri agafe nke ọma ma na-edobe TMP n'ime windo siri ike. Site n'ime nke a, ndị na-arụ ọrụ na-ebelata polarization na ntụpọ akpụkpọ ahụ, na-ebelata ma ugboro ole a ga-eji dochie akpụkpọ ahụ na usoro nhicha - na-ebelata ọnụ ahịa ọrụ ozugbo ma na-eme ka mmepụta ngwaahịa ka mma.

Mgbanwe na nlekota kwesịrị ekwesị nke mgbanwe ndị a—gụnyere nha viscosity nke ngwọta protein n'oge a—dị mkpa iji melite arụmọrụ ntinye ultrafiltration na ibelata mmetụta ọjọọ metụtara polarization nke ntinye na nhazi protein.

Imeziwanye Ultrafiltration maka Ngwọta Protein Dị Elu

6.1. Usoro Kachasị Mma n'Ọrụ

Ijigide arụmọrụ ultrafiltration kacha mma site na iji ngwọta protein viscosity dị elu chọrọ nguzozi dị nro n'etiti nrụgide transmembrane (TMP), ntinye protein, na viscosity ngwọta. TMP—ọdịiche dị na nrụgide n'ofe akpụkpọ ahụ—na-emetụta ọsọ ntinye protein ultrafiltration ozugbo na ogo ntụpọ akpụkpọ ahụ. Mgbe ị na-edozi ngwọta viscous dị ka ọgwụ mgbochi monoclonal ma ọ bụ protein serum centric concentration dị elu, mmụba dị ukwuu ọ bụla na TMP nwere ike ibuli flux na mbụ, mana ọ na-emekwa ngwa ngwa ka ntụpọ na mkpokọta protein dị n'elu akpụkpọ ahụ. Nke a na-eduga na usoro nzacha na-adịghị mma na nke na-adịghị akwụsi ike, nke emere site na nyocha onyonyo na-egosi akwa protein dị oke njọ na-etolite na TMP dị elu na mkpokọta protein karịrị 200 mg/mL.

Ụzọ kachasị mma isi mee ya bụ ịgba sistemụ ahụ nso, mana ọ bụghị ịgafe, TMP dị oke mkpa. N'oge a, a na-eme ka mmepụta ya dịkwuo elu mana ihe egwu nke mmebi a na-apụghị ịgbanwe agbanwe ka dị obere. Maka oke viscosities, nchọpụta ndị na-adịbeghị anya na-egosi mbelata TMP na ịbawanye mmiri nri (nzacha transverse flow) n'otu oge iji nyere aka belata polarization na ntinye protein. Dịka ọmụmaatụ, ọmụmụ ihe na Fc-fusion protein concentration na-egosi obere ntọala TMP na-enyere aka ịnọgide na-enwe mmiri kwụsiri ike ma na-ebelata mfu ngwaahịa.

Mmụba nke nta nke nta na usoro nke protein n'oge ultrafiltration dị oke mkpa. Usoro ntinye ngwa ngwa nwere ike ịmanye ngwọta ahụ ka ọ bụrụ usoro viscosity dị elu ngwa ngwa, na-eme ka ihe egwu nchịkọta na oke nke mmebi ahụ dịkwuo elu. Kama nke ahụ, ịbawanye ọkwa protein na-enye ohere ka a gbanwee paramita usoro dịka TMP, ọsọ cross-flow, na pH n'otu oge, na-enyere aka ịnọgide na-enwe nkwụsi ike sistemụ. Ọmụmụ ihe gbasara enzyme ultrafiltration na-akwado na ịnọgide na-enwe obere nrụgide ọrụ n'oge usoro ndị a na-eme ka mmụba a na-achịkwa na ntinye, na-ebelata mbelata flux ebe a na-echebe iguzosi ike n'ezi ihe nke ngwaahịa.

6.2. Ugboro na Mmezi Mgbanwe Akpụkpọ ahụ

Ugboro ole a na-eji dochie akpụkpọ ahụ na ultrafiltration jikọtara ya na ihe ngosi nke mmebi na mbelata flux. Kama ịdabere naanị na mbelata flux dị ka ihe ngosi njedebe nke ndụ, ileba anya na iguzogide ntụpọ kpọmkwem - ihe nha nha nke na-anọchite anya iguzogide nke ihe ekpokọtara na-eweta - egosila na a pụrụ ịtụkwasị ya obi karịa, ọkachasị na nri protein ma ọ bụ protein-polysaccharide agwakọtara, ebe mmebi nwere ike ime ngwa ngwa na nke siri ike.

Nleba anya maka ihe ngosi mmebi ndị ọzọ dịkwa oke mkpa. Ihe ịrịba ama a na-ahụ anya nke itinye n'elu, mmiri na-adịghị agbanwe agbanwe, ma ọ bụ mmụba na-aga n'ihu na TMP (n'agbanyeghị nhicha) bụ ihe mgbaàmà ịdọ aka ná ntị nke mmebi dị elu nke na-ebute ọdịda akpụkpọ ahụ. Usoro dịka ịchụso ihe ndekọ mmebi agbanwere (MFI-UF) na ijikọ ya na arụmọrụ akpụkpọ ahụ na-eme ka usoro nhazi amụma nke nnọchi anya kama mgbanwe mmeghachi omume, si otú a na-ebelata oge nkwụsị na ijikwa ụgwọ mmezi.

Ọ bụghị naanị site na mkpokọta ihe ndị na-emebi ihe na-emebi ihe na akpụkpọ ahụ kamakwa site na nchara, ọkachasị n'usoro na-agba ọsọ na oke pH ma ọ bụ nwere nnukwu nnu. Ekwesịrị ịme nyocha mgbe niile na usoro nhicha kemịkalụ iji jikwaa ma ihe ndị na-emebi ihe na ihe ndị na-emebi ihe. Mgbe a hụrụ ihe ndị metụtara nchara, a ga-agbanwerịrị ugboro ole a na-ehicha akpụkpọ ahụ na oge nnọchi iji hụ na akpụkpọ ahụ na-adịgide adịgide na arụmọrụ ultrafiltration membrane na-aga n'ihu. Nlekọta zuru oke na nke a haziri ahazi dị mkpa iji belata mmetụta nke nsogbu ndị a na ịgbatị ọrụ dị irè.

6.3. Njikwa Usoro na Nha Viscosity Inline

Nnwale ziri ezi, nke oge a na-eme n'ezie nke viscosity nke ngwọta protein dị mkpa maka njikwa usoro na ultrafiltration, karịsịa ka njupụta na viscosities na-abawanye. Sistemụ nha viscosity n'ime ahịrị na-enye nlekota na-aga n'ihu, na-enye ohere nzaghachi ozugbo ma na-eme ka mgbanwe dị iche iche na paramita sistemụ dị ike.

Teknụzụ ndị na-apụta ọhụrụ agbanweela ọdịdị ala nke nha viscosity nke ngwọta protein:

Raman Spectroscopy na Kalman Filter: Nyocha Raman n'oge a, nke ihe nzacha Kalman gbasaara na-akwado, na-enye ohere ịchọpụta mkpokọta protein na nhazi buffer siri ike. Ụzọ a na-eme ka mmetụta na izi ezi dịkwuo elu, na-akwado akpaaka usoro maka ntinye ultrafiltration na diafiltration.

Viscometry Kinematic Capillary Akpaaka: Site na iji ọhụụ kọmputa, teknụzụ a na-atụle viscosity ngwọta na akpaghị aka, na-emeri njehie aka ma na-enye nlekota ugboro ugboro, nke a na-emegharị ugboro ugboro n'ọtụtụ usoro. A kwadoro ya maka ma nhazi protein ọkọlọtọ na nke dị mgbagwoju anya ma na-ebelata ntinye aka n'oge usoro ntinye ultrafiltration.

Ngwaọrụ Ọmụmụ Ihe Ndị Dị na Microfluidic: Sistemụ microfluidic na-enye nkọwa zuru ezu, na-aga n'ihu, ọbụlagodi maka ngwọta protein ndị na-abụghị nke Newtonian, nke nwere oke viscosity. Ndị a bara uru karịsịa na mmepụta ọgwụ, na-akwado atụmatụ teknụzụ nyocha usoro (PAT) na ijikọta ya na loops nzaghachi.

Njikwa usoro site na iji ngwaọrụ ndị a na-eme ka mmejuputa loops nzaghachi maka mgbanwe oge TMP, ọnụego nri, ma ọ bụ ọsọ ngafe na nzaghachi maka mgbanwe viscosity. Dịka ọmụmaatụ, ọ bụrụ na mmetụta inline achọpụta mmụba mberede na viscosity (n'ihi mmụba ma ọ bụ nchịkọta), enwere ike ibelata TMP na akpaghị aka ma ọ bụ bulie ọsọ ngafe iji belata mmalite nke polarization nke njupụta na ultrafiltration. Ụzọ a abụghị naanị ịgbatị ndụ membrane kamakwa ọ na-akwado ịdị mma ngwaahịa na-aga n'ihu site na ijikwa ihe ndị na-emetụta viscosity nke ngwọta protein n'ụzọ dị iche iche.

Nhọrọ nke teknụzụ nlekota viscosity kacha mma dabere na ihe achọrọ kpọmkwem nke ngwa ultrafiltration, gụnyere oke viscosity a tụrụ anya ya, mgbagwoju anya nke nhazi protein, mkpa njikọta, na ọnụ ahịa. Mmepe ndị a na nlekota oge na njikwa usoro ike emeela ka ikike imeziwanye ultrafiltration maka ngwọta protein viscosity dị elu, na-ahụ na ma arụmọrụ kwụsiri ike ma mmepụta ngwaahịa dị elu.

Nchọpụta nsogbu na nsogbu ndị a na-ahụkarị na nzacha protein Ultrafiltration

7.1. Mgbaàmà, Ihe Ndị Na-akpata Ya, na Ọgwụgwọ Ya

Mmụba Nrụgide Transmembrane

Mmụba nke nrụgide transmembrane (TMP) n'oge ultrafiltration na-egosi mmụba nke iguzogide n'ofe akpụkpọ ahụ. Mmetụta nke nrụgide transmembrane na ultrafiltration bụ kpọmkwem: oke nrụgide transmembrane nkịtị na-adaberekarị na usoro, mana mmụba na-adịgide adịgide kwesịrị ka a nyochaa ya. Ihe abụọ a na-ahụkarị pụtara ìhè:

Ọbara dị elu nke mmiri protein:Ka mmiri protein na-agbakọta nke ukwuu—nke a na-ahụkarị n'ọkwa protein ultrafiltration dị elu—nrụgide dị mkpa maka mmiri na-arị elu. Nke a na-apụta ìhè na usoro ntinye na diafiltration ikpeazụ ebe mmiri protein na-agbapụta nke ọma.
Mmebi akpụkpọ ahụ:Ihe ndị na-eme ka ihe dị ka protein aggregates ma ọ bụ ngwakọta protein polysaccharide nwere ike ịrapara ma ọ bụ gbochie oghere akpụkpọ ahụ, na-eme ka TMP gbapụta ngwa ngwa.

Ọgwụgwọ:

Belata TMP ma mụbaa ọsọ nri: Ibelata TMP ebe ọ na-eme ka ọsọ nri dịkwuo elu na-ebelata polarization nke njupụta na nhazi oyi akwa gel, na-akwalite mmụba kwụsiri ike.
Ịsacha akpụkpọ ahụ mgbe niile: Mepụta ugboro kachasị mma iji hichaa akpụkpọ ahụ iji wepụ ihe ndị na-emebi emebi. Lelee irè ya site na nha nha protein solution mgbe e sachara ya.
Dochie akpụkpọ ahụ ndị merela agadi: Ọ ga-adị mkpa ka e mee ka ugboro ugboro nke mkpuchi akpụkpọ ahụ dịkwuo mma ma ọ bụrụ na nhicha ahụ ezughi oke ma ọ bụ na ogologo oge nke akpụkpọ ahụ erutela.

Ọnụego Mgbanwe Na-ebelata: Osisi Nchọpụta

Mbelata mgbe niile na mmiri na-agbagharị n'oge usoro ntinye ultrafiltration na-egosi nsogbu mmepụta ihe. Soro usoro nchọpụta a:

Nyochaa TMP na viscosity:Ọ bụrụ na ha abụọ amụbaala, lelee ma ọ bụrụ na ha nwere ntụpọ ma ọ bụ gel oyi akwa.
Lelee ihe mejupụtara nri na pH:Mgbanwe ebe a nwere ike ịgbanwe viscosity nke ngwọta protein ma kwalite mmebi.
Nyochaa arụmọrụ nke membrane:Mbelata na mmiri na-asọpụta n'agbanyeghị nhicha na-egosi na mmebi akpụkpọ ahụ nwere ike ime ma ọ bụ mmebi a na-apụghị ịgbanwe agbanwe.

Ngwọta:

Mee ka okpomọkụ, pH, na ike ionic dịkwuo mma na nri iji belata mmebi na polarization nke ihe ndị dị n'ime ya na ultrafiltration.
Jiri modulu akpụkpọ ahụ e megharịrị n'elu ma ọ bụ nke na-agbagharị agbagharị iji mebie akwa gel ma weghachi mmiri na-agbagharị.
Mee nha nha protein solution iji tụọ mgbanwe ndị na-emetụta mmiri.

Mkpụcha akpụkpọ ahụ ngwa ngwa ma ọ bụ gel

Mmalite nke oyi akwa jel ngwa ngwa na-esi na oke polarization nke ukwuu na elu akpụkpọ ahụ pụta. Nrụgide transverse flow filtration (TFF) transmembrane na-adịkarị mfe karịsịa n'okpuru ọnọdụ nri dị elu ma ọ bụ nke nwere nnukwu protein.

Atụmatụ Mbelata:

Tinye elu akpụkpọ ahụ nwere ike ịdị ọcha ma na-acha ọkụ nke ọma (dịka ọmụmaatụ, akpụkpọ ahụ Polyvinylidene fluoride [PVDF]) iji belata njikọ protein na njikọ ya.
Jiri coagulation ma ọ bụ electrocoagulation gwọọ nri tupu oge eruo iji wepụ ihe ndị na-emerụ ahụ nke ukwuu tupu a na-eme ya nke ọma.
Tinye ngwaọrụ igwe dịka modulu na-agbagharị agbagharị n'ime usoro nzacha transverse flow iji belata ọkpụrụkpụ oyi akwa achịcha ma belata nhazi oyi akwa gel.

7.2. Ịgbanwe ka mgbanwe nri si dị

Sistemụ nzacha protein ultrafiltration ga-agbanwerịrị ka ọ dị iche na njirimara ma ọ bụ nhazi protein nri. Ihe ndị na-emetụta viscosity nke ngwọta protein - dịka nhazi buffer, ntinye protein, na oke mkpokọta - nwere ike ịgbanwe omume sistemụ.

Atụmatụ Nzaghachi

Nlekota na nhazi nke oge a:Tinye ihe mmetụta nyocha n'ịntanetị (Raman spectroscopy + Kalman filtering) maka nchọpụta ngwa ngwa nke mgbanwe nri, na-arụ ọrụ karịa usoro UV ma ọ bụ IR ochie.
Njikwa usoro mgbanwe:Gbanwee ntọala paramita (ọnụego eruba, TMP, nhọrọ akpụkpọ ahụ) dịka nzaghachi nye mgbanwe achọpụtara. Dịka ọmụmaatụ, mmụba nke viscosity protein solution nwere ike ịchọ obere TMP na oke nha nha.
Nhọrọ akpụkpọ ahụ:Jiri akpụkpọ ahụ nwere nha oghere na kemịkalụ elu nke e mere ka ọ dị mma maka ihe oriri dị ugbu a, na-edozi njigide protein na mmụba ya.
Nri tupu ọgwụgwọ:Ọ bụrụ na mgbanwe mberede na nri na-eme ka ọ ghara ịdị mma, tinye usoro ịcha ọbara ma ọ bụ nzacha tupu ultrafiltration amalite.

Ihe atụ:

Na bioprocessing, mgbanwe nchekwa ma ọ bụ mgbanwe na mkpokọta antibody kwesịrị ịkpalite mgbanwe TMP na usoro mmiri site na sistemụ njikwa.
Maka ultrafiltration jikọtara na chromatography, algọridim nhazi ngwakọta-integer nwere ike ibelata mgbanwe ma belata ọnụ ahịa ọrụ ebe ọ na-ejigide arụmọrụ membrane ultrafiltration.

Nlekota nha viscosity nke protein solution na nhazi ozugbo na ọnọdụ usoro na-enyere aka ime ka ntinye ultrafiltration dịkwuo mma, na-eme ka mmepụta ya dị mfe, ma belata mmebi akpụkpọ ahụ na polarization nke njupụta.

Ajụjụ Ndị A Na-ajụkarị

8.1. Kedu oke nkịtị maka nrụgide transmembrane na ultrafiltration nke ngwọta protein?

Oke nrụgide transmembrane nkịtị (TMP) na sistemụ ntinye protein ultrafiltration na-adabere n'ụdị akpụkpọ ahụ, nhazi modulu, na njirimara nri. Maka ọtụtụ usoro ultrafiltration protein, a na-edobe TMP n'etiti 1 ruo 3 bar (15–45 psi). Ụkpụrụ TMP karịrị 0.2 MPa (ihe dị ka 29 psi) nwere ike ibute mmebi akpụkpọ ahụ, mmebi ngwa ngwa, na ndụ akpụkpọ ahụ dị mkpụmkpụ. Na ngwa biomedical na bioprocessing, TMP akwadoro ekwesịghị ịgafe 0.8 bar (~12 psi) iji zere mgbawa akpụkpọ ahụ. Maka usoro dị ka nzacha mmiri transverse, ịnọ n'ime oke TMP a na-echebe ma mmepụta na iguzosi ike n'ezi ihe protein.

8.2. Kedu ka viscosity nke ngwọta protein si emetụta arụmọrụ ultrafiltration?

Ọbara ọbara nke ihe ngwọta protein na-emetụta arụmọrụ nke oke mmiri ultrafiltration. Ọbara ọbara nke na-eme ka iguzogide mmiri dịkwuo elu ma na-eme ka TMP dịkwuo elu, na-ebute mbelata mmiri ọbara nke na-abanye n'ime ahụ na mmebi akpụkpọ ahụ ngwa ngwa. A na-egosipụta mmetụta a site na ọgwụ mgbochi monoclonal ma ọ bụ protein Fc-fusion na oke mmiri, ebe viscosity na-abawanye n'ihi mmekọrịta protein-protein na mmetụta chajị. Ijikwa na imeziwanye viscosity na ihe ndị na-enyere aka ma ọ bụ ọgwụgwọ enzymatic na-eme ka flux dịkwuo mma, na-ebelata ntụpọ, ma na-enye ohere ka oke mmiri ahụ nwee ike iru n'oge usoro ntinye ultrafiltration. Nlekota nha viscosity nke ihe ngwọta protein dị oke mkpa maka ịnọgide na-enwe nhazi dị irè.

8.3. Gịnị bụ polarization nke njupụta na ihe kpatara o ji dị mkpa na TFF?

Mkpokọta mkpụrụ ndụ ihe nketa na ultrafiltration bụ mkpokọta protein na elu akpụkpọ ahụ, na-eme ka ọ dị larịị n'etiti ihe ngwọta buru ibu na njikọ akpụkpọ ahụ. Na nzacha mmiri transverse, nke a na-eduga na mmụba nke viscosity mpaghara na mbelata nke flux nwere ike ịgbanwe agbanwe. Ọ bụrụ na ejighị ya, ọ nwere ike ịkwalite ntụpọ akpụkpọ ahụ ma belata arụmọrụ sistemụ. Ijikwa polarization n'ime ultrafiltration gụnyere imeziwanye ọnụego mmiri cross-flow, TMP, na nhọrọ akpụkpọ ahụ iji nọgide na-enwe obere polarization. Njikwa ziri ezi na-eme ka mmepụta dị elu ma nwee obere ihe egwu nke foulating.

8.4. Kedu ka m ga-esi kpebie mgbe m ga-edochi akpụkpọ ahụ m nke na-anaghị amịpụta ihe?

Dochie akpụkpọ ahụ ultrafiltration mgbe ị hụrụ mbelata dị ukwuu na mmepụta (flux), mmụba na-aga n'ihu na TMP nke nhicha ọkọlọtọ enweghị ike idozi, ma ọ bụ mmetọ a na-ahụ anya nke na-anọgide mgbe e hichara ya. Ihe ngosi ndị ọzọ gụnyere mfu nke nhọrọ (enweghị ịjụ protein ndị a na-elekwasị anya dịka a tụrụ anya ya) na enweghị ike iru nkọwapụta arụmọrụ. Nlekota ugboro mgbanwe akpụkpọ ahụ site na iji nnwale mgbanwe na nhọrọ mgbe niile bụ ntọala maka ime ka ogologo ndụ akpụkpọ ahụ dịkwuo elu na usoro ntinye protein ultrafiltration.

8.5. Kedu paramita ọrụ m nwere ike ịgbanwe iji belata mmebi protein na TFF?

Isi ihe dị mkpa iji belata mmebi protein na nzacha transverse flow gụnyere:

Nọgide na-enwe ọsọ zuru oke iji belata mkpokọta protein mpaghara ma jikwaa polarization nke mkpokọta.
Rụọ ọrụ n'ime oke TMP akwadoro, nke na-abụkarị 3-5 psi (0.2–0.35 bar), iji gbochie oke mmiri mmiri na mmebi akpụkpọ ahụ.
Tinye usoro nhicha akpụkpọ ahụ mgbe niile iji belata mmebi a na-apụghị ịgbagha agbagha.
Lelee ma, ọ bụrụ na ọ dị mkpa, mee ka mmiri nri ahụ dịgide tupu oge eruo iji chịkwaa viscosity (dịka ọmụmaatụ, site na iji ọgwụgwọ enzymatic dị ka pectinase).
Họrọ ihe mkpuchi akpụkpọ ahụ na nha oghere (MWCO) dabara adaba maka nha protein na ebumnuche nhazi.

Ijikọta hydrocyclone prefiltration ma ọ bụ ọgwụgwọ enzymatic pre-treatment nwere ike ime ka arụmọrụ sistemụ ka mma, ọkachasị maka nri nwere oke viscosity. Soro usoro nhazi nri nke ọma ma gbanwee ntọala ya nke ọma iji belata mmebi akpụkpọ ahụ ma melite usoro ntinye ultrafiltration.