Ukulawula ukujiya kwezixazululo zamaprotheni kubalulekile ekwenzeni ngcono izinqubo zokujiya kwe-ultrafiltration ekukhiqizweni kwe-biopharmaceutical. Ukujiya okuphezulu kwezixazululo zamaprotheni—ikakhulukazi ekujiya okuphezulu kwamaprotheni—kuthinta ngqo ukusebenza kwe-membrane, ukusebenza kahle kwenqubo, kanye nomnotho ekusetshenzisweni kokujiya kwe-protein kwe-ultrafiltration. Ukujiya kwesixazululo kukhuphuka ngokuqukethwe kwamaprotheni ngenxa yokuhlanganiswa kwe-anti-body kanye nokusebenzisana kwe-electrostatic, okwandisa ukumelana nokugeleza kanye nokwehla kwengcindezi kulo lonke ulwelwesi lwe-ultrafiltration. Lokhu kuphumela ekujuleni okuphansi kwe-permeate kanye nezikhathi zokusebenza ezinde, ikakhulukazi ezinqubweni zokuhlunga ukugeleza okuguquguqukayo (TFF).
Ingcindezi ye-Transmembrane (TMP), amandla aqhubayo ngemuva kokuhluzwa kwe-ultramembrane, ixhumene kakhulu ne-viscosity. Ukusebenza ngaphandle kobubanzi obujwayelekile bengcindezi ye-transmembrane kusheshisa ukungcoliswa kwe-membrane futhi kukhulise ukuhluzwa kwe-concentration—ukuqongelela kwamaprotheni eduze kwe-membrane okwandisa njalo ukuhluzwa kwendawo. Kokubili ukuhluzwa kwe-concentration kanye nokuhluzwa kwe-membrane kuholela ekusebenzeni okunciphile kwe-ultrafiltration membrane futhi kunganciphisa isikhathi sokuphila kwe-membrane uma kungahlolwa. Umsebenzi wokuhlola ukhombisa ukuthi ukungcoliswa kwe-membrane kanye nokuhluzwa kwe-concentration ku-ultrafiltration kugqama kakhulu ngamanani aphezulu e-TMP kanye nokuphakelwa okunamandla kakhulu, okwenza ukulawulwa kwe-TMP kwesikhathi sangempela kubalulekile ukuze kukhuliswe ukugeleza futhi kuncishiswe imvamisa yokuhlanza.
Ukuthuthukisa ukuhlushwa kwe-ultrafiltration kudinga amasu ahlanganisiwe:
- Ukulinganiswa kwe-viscosity yesisombululo seprotheni: Ukuhlolwa kwe-viscosity okuvamile—kusetshenziswaama-viscometer asemgqeni—siza ukubikezela amazinga okuhlunga futhi silindele izithiyo zenqubo, sisekele ukuguqulwa kwenqubo okusheshayo.
- Ukulungisa ukudla: Ukulungisa i-pH, amandla e-ionic, kanye nokushisa kunganciphisa i-viscosity futhi kunciphise ukungcola. Isibonelo, ukwengeza ama-ion e-sodium kuthuthukisa ukuxoshwa kwamanzi phakathi kwamaprotheni, ukunciphisa ukuhlangana kanye nokungcola, kuyilapho ama-ion e-calcium evame ukukhuthaza ukuhlangana kanye nokungcola kwamaprotheni.
- Ukusetshenziswa kwezinto ezisizayo: Ukufaka izinto ezisiza ukwehlisa i-viscosity ezixazululweni zamaprotheni agxile kakhulu kuthuthukisa ukungena kwe-membrane futhi kunciphisa ingcindezi ye-transmembrane ekuhlungeni kwe-ultrafiltration, okwandisa ukusebenza kahle okuphelele.
- Izindlela zokugeleza ezithuthukisiwe: Ukwandisa ijubane lokuhamba kwe-cross-flow, ukusebenzisa ukugeleza kwe-cross-flow okushintshanayo, noma ukusebenzisa umjovo we-air jet kuphazamisa izendlalelo zokungcola. Lezi zindlela zisiza ukugcina ukugeleza kwe-permeating futhi zinciphise imvamisa yokufakwa kwe-membrane ngokunciphisa ukwakheka kwe-deposit.
- Ukukhetha nokuhlanza ulwelwesi: Ukukhetha ama-membrane aqinile ngamakhemikhali (isb., i-SiC noma ama-thermosalient hybrids) kanye nokwenza ngcono imvamisa yokuhlanza ama-membrane ngezinqubo ezifanele (isb., ukuhlanza i-sodium hypochlorite) kubalulekile ekwandiseni isikhathi sokuphila kwama-membrane kanye nokunciphisa izindleko zokusebenza.
Sekukonke, ukulawulwa kwe-viscosity okuphumelelayo kanye nokuphathwa kwe-TMP kuyitshe lesisekelo lokusebenza kwesigaba sokuhluzwa kwe-ultrafiltration okuphumelelayo, okuthonya ngqo ukukhiqizwa komkhiqizo, imvamisa yokuhlanza i-membrane, kanye nobude bempahla ye-membrane ebizayo.
Ukuqonda Ukuqina Kwesisombululo Samaprotheni ku-Ultrafiltration
1.1. Kuyini Ukuqina Kwezixazululo Zamaphrotheni?
I-Viscosity ichaza ukumelana koketshezi ekugelezeni; ezixazululweni zamaprotheni, iphawula ukuthi ukungqubuzana kwama-molecule kuphazamisa kangakanani ukunyakaza. Iyunithi ye-SI ye-viscosity yi-Pascal-second (Pa·s), kodwa i-centipoise (cP) ivame ukusetshenziswa kuma-biological fluids. I-Viscosity ithinta ngqo indlela izixazululo zamaprotheni ezingapontshwa kalula ngayo noma zihlungwe ngesikhathi sokukhiqiza futhi ithinta ukulethwa kwemithi, ikakhulukazi kuma-biotherapeutics agxile kakhulu.
Ukuhlushwa kwamaprotheni kuyisici esiyinhloko esithonya i-viscosity. Njengoba amazinga amaprotheni ekhuphuka, ukusebenzisana phakathi kwama-molecule kanye nokuminyana kuyanda, okubangela i-viscosity ikhuphuke, ngokuvamile ngokungaqondile. Ngaphezu komkhawulo othile, ukusebenzisana kwamaprotheni nama-protein kucindezela kakhulu ukusabalala ngaphakathi kwesisombululo. Isibonelo, izixazululo ze-antibody ezihlanganisiwe ze-monoclonal ezisetshenziswa emithini zivame ukufinyelela amazinga e-viscosity aphonsela inselelo umjovo ongaphansi kwesikhumba noma anciphise amazinga okucubungula.
Amamodeli abikezela i-viscosity ezixazululweni zamaprotheni agxilile manje afaka i-geometry yama-molecule kanye nokuthambekela kokuhlangana. Ukwakheka kwamaprotheni—kungakhathaliseki ukuthi kunde, kuyindilinga, noma kuthambekele ekuhlanganeni—kuthinta kakhulu i-viscosity ekugxilweni okuphezulu. Intuthuko yakamuva ekuhlolweni kwe-microfluidic ivumela ukulinganiswa okunembile kwe-viscosity kusuka kumanani amancane esampula, okwenza kube lula ukuhlolwa okusheshayo kwamafomula amasha amaprotheni.
1.2. Indlela Ukuqina Okushintsha Ngayo Ngesikhathi Sokuhlunga Okuphezulu
Ngesikhathi sokuhluzwa kwe-ultrafiltration, i-concentration polarization iqoqa ngokushesha amaprotheni esibonakalayo se-membrane-solution. Lokhu kudala ama-gradients amakhulu okuhluzwa kwendawo futhi kuphakamisa i-viscosity eduze kwe-membrane. I-viscosity ephakeme kulesi sifunda ivimbela ukudluliselwa kwesisindo futhi inciphise ukuhamba kwamanzi.
Ukuhlukaniswa kokuhlushwa kuhlukile ekungcolisweni kwe-membrane. Ukuhlukaniswa kuyashintshashintsha futhi kuyashintshashintsha, kwenzeka ngemizuzu embalwa njengoba ukuhlunga kuqhubeka. Uma kuqhathaniswa, ukungcoliswa kuyaqhubeka ngokuhamba kwesikhathi futhi kuvame ukuhilela ukufakwa okungenakuguqulwa noma ukuguqulwa kwamakhemikhali ebusweni be-membrane. Ukuxilongwa okunembile kuvumela ukulandelwa kwesikhathi sangempela kwengqimba yokuhlushwa kokuhlushwa, kwembula ukuzwela kwayo ejubaneni lokuhamba kanye nokucindezela kwe-transmembrane. Isibonelo, ukwandisa ijubane noma ukunciphisa ingcindezi ye-transmembrane (TMP) kusiza ukuphazamisa ungqimba lomngcele oqinile, ukubuyisela ukugeleza.
Amapharamitha okusebenza athinta ngqo ukuziphatha kwe-viscosity:
- Ingcindezi ye-Transmembrane (TMP): I-TMP ephezulu ikhulisa ukuvuleka, ikhuphule ukujiya kwendawo futhi yehlise ukugeleza.
- Ijubane lokugeleza okuwelayo: Ijubane elithuthukisiwe linciphisa ukuqongelela, lilinganisela ukujiya eduze kwe-membrane.
- Imvamisa yokuhlanza ulwelwesi: Ukuhlanza njalo kunciphisa ukwakheka kwesikhathi eside futhi kunciphisa ukulahleka kokusebenza okubangelwa ukuchachamba.
Izigaba zokuhlushwa kwe-Ultrafiltration kumele zithuthukise lezi zinhlaka ukuze kuncishiswe imiphumela emibi ye-viscosity futhi kugcinwe ukuphuma kwayo.
1.3. Izakhiwo Zesixazululo Samaprotheni Ezithinta Ukujiya
Isisindo sama-moleculefuthiukwakhekaikakhulukazi nquma ukujiya. Amaprotheni amakhulu, ayinkimbinkimbi noma ama-aggregate akhiqiza ukujiya okuphezulu ngenxa yokunyakaza okuvimbekile kanye namandla amakhulu aphakathi kwama-molecule. Ukuma kwamaprotheni kushintsha kakhulu ukugeleza—amaketanga amade noma athambekele ekuhlanganisweni abangela ukumelana okukhulu kunamaprotheni ahlanganisiwe e-globular.
pHKuthinta kakhulu ukushaja kweprotheyini kanye nokuncibilika kwayo. Ukulungisa i-pH yesisombululo eduze kwephuzu le-isoelectric leprotheyini kunciphisa ukushaja okune-net, kunciphisa ukuxoshwa kweprotheyini neprotheyini, futhi kunciphisa i-viscosity okwesikhashana, okwenza kube lula ukuhlunga. Isibonelo, ukusebenzisa i-ultrafiltration eduze kwephuzu le-isoelectric le-BSA noma i-IgG kungathuthukisa kakhulu ukugeleza kwe-permeate kanye nokukhetha kokuhlukaniswa.
Amandla e-Ionickuthinta i-viscosity ngokushintsha ungqimba oluphindwe kabili lukagesi oluzungeze amaprotheni. Amandla e-ionic anda ahlola ukusebenzisana kwe-electrostatic, akhuthaze ukudluliselwa kwamaprotheni ngama-membrane kodwa futhi akhuphula ingozi yokuhlangana kanye nokuphakama kwe-viscosity okuhambisanayo. Ukuhwebelana phakathi kokusebenza kahle kokudlulisela kanye nokukhetha kuvame ukuncika ekulungiseni kahle amazinga kasawoti kanye nokwakheka kwe-buffer.
Izithasiselo ezincane zama-molecule—njenge-arginine hydrochloride noma i-guanidine—zingasetshenziswa ukunciphisa i-viscosity. Lezi zinto ziphazamisa ukukhanga kwe-hydrophobic noma i-electrostatic, zinciphisa ukuhlangana, futhi zithuthukise izakhiwo zokugeleza kwesisombululo. Izinga lokushisa lisebenza njengenguquko yokulawula eyengeziwe; amazinga okushisa aphansi andisa i-viscosity, kuyilapho ukushisa okwengeziwe kuvame ukuyinciphisa.
Ukulinganiswa kwe-viscosity yesisombululo seprotheni kufanele kucabangele:
- Ukusatshalaliswa kwesisindo sama-molecule
- Ukwakheka kwesisombululo (usawoti, izinto ezisizayo, izithasiselo)
- Ukukhetha i-pH kanye nesistimu ye-buffer
- Ukusetha amandla e-Ionic
Lezi zici zibalulekile ekwenzeni ngcono ukusebenza kwe-membrane ye-ultrafiltration kanye nokuqinisekisa ukuvumelana kuzo zonke izigaba zokuhlushwa kanye nezinqubo ze-TFF.
Izisekelo Zokuhlushwa Kwamaprotheni Okuhlushwa Kwe-Ultrafiltration
Izimiso Zesigaba Sokuhlushwa Kwe-Ultrafiltration
Ukuhlushwa kwamaprotheni e-Ultrafiltration kusebenza ngokusebenzisa ingcindezi ye-transmembrane (TMP) kulwelwesi olungena kancane, kuqhube i-solvent kanye ne-solutes encane ngenkathi kugcinwa amaprotheni nama-molecule amakhulu. Le nqubo isebenzisa ukuhlushwa okukhethiwe okusekelwe kusayizi wama-molecule, lapho i-membrane's molecular weight cut-off (MWCO) ichaza usayizi omkhulu wama-molecule adlulayo. Amaprotheni adlula i-MWCO aqoqana ohlangothini olungenawo umswakama, andise ukuhlushwa kwawo njengoba ukugcwala kuhoxiswa.
Isigaba sokuhlushwa kwe-ultrafiltration sihlose ukunciphisa ivolumu kanye nokucebisa isisombululo seprotheyini. Njengoba ukuhlunga kuqhubeka, i-viscosity yesisombululo seprotheyini ivame ukukhuphuka, kuthinte ukugeleza kanye nezidingo ze-TMP. Amaprotheni agciniwe angase asebenzisane futhi asebenzisane ne-membrane, okwenza inqubo yangempela ibe yinkimbinkimbi kakhulu kunokungafakwa kalula kosayizi. Ukusebenzisana kwe-electrostatic, ukuhlanganiswa kwamaprotheni, kanye nezici zesisombululo ezifana ne-pH kanye namandla e-ionic kuthinta imiphumela yokugcina kanye nokuhlukanisa. Kwezinye izimo, ukuthuthwa kwe-advective kubusa ngaphezu kokusabalalisa, ikakhulukazi kuma-membrane anezimbobo ezinkulu, okwenza kube nzima ukulindela ngokusekelwe kuphela ekukhethweni kwe-MWCO [bheka isifinyezo socwaningo].
Incazelo Yokuhlunga Ukugeleza Okuguquguqukayo (i-TFF)
Ukuhlunga ukugeleza okuguquguqukayo, okubizwa nangokuthi ukuhlunga ukugeleza okuguquguqukayo (i-TFF), kuhambisa isisombululo seprotheyini ngendlela ejikelezayo ngaphesheya kobuso be-membrane. Le ndlela ihluke ekuhlungeni okungapheli, lapho ukugeleza kuqondile ku-membrane, kusunduza izinhlayiya ngqo ziye esihlungini.
Umehluko obalulekile kanye nemiphumela:
- Ukulawula Ukungcola:I-TFF inciphisa ukunqwabelana kwamaprotheni kanye nezinhlayiya, okwaziwa ngokuthi ukwakheka kwekhekhe, ngokuqhubeka nokukhuculula izinto ezingaba khona eziphuma kulwelwesi. Lokhu kuphumela ekugelezeni kokungena kwamanzi okuzinzile kanye nokugcinwa okulula.
- Ukugcinwa Kwamaprotheni:I-TFF isekela ukuphathwa okungcono kwe-concentration polarization—ungqimba lwama-molecule agciniwe eduze kwe-membrane—okungathi uma ingalawulwa, inganciphisa ukukhetha kokuhlukaniswa futhi ithuthukise ukungcola. Ukugeleza okuguquguqukayo ku-TFF kunciphisa lo mphumela, kusiza ukugcina ukugcinwa kwamaprotheni amaningi kanye nokusebenza kahle kokuhlukaniswa.
- Ukuzinza kwe-Flux:I-TFF ivumela isikhathi eside sokusebenza ngokuguquguquka okuqhubekayo, okwandisa ukusebenza kahle ezinqubweni ezinamaprotheni amaningi noma ukudla okucebile ngezinhlayiya. Ngokuphambene, ukuhlunga okungapheli kuphazanyiswa ngokushesha ukungcola, ukwehlisa ukuphuma kwegazi futhi kudinga ukungenelela kokuhlanza njalo.
Izinhlobo ze-TFF ezithuthukisiwe, njengokugeleza kwe-tangential okushintshanayo (i-ATF), ziphazamisa kakhulu ukungcolisa kanye nokwakheka kwekhekhe ngokuguqula noma ukushintsha ijubane le-tangential ngezikhathi ezithile, kwandise isikhathi sokuphila kwesihlungi kanye nokuthuthukisa ukugeleza kwamaprotheni [bheka isifinyezo socwaningo]. Kokubili ukusethwa kwe-TFF yakudala kanye nethuthukisiwe, izilungiselelo zokusebenza—njenge-TMP, ijubane lokuhamba kwe-crossflow, kanye nemvamisa yokuhlanza—kumele kulungiselelwe uhlelo oluthile lwamaprotheni, uhlobo lwe-membrane, kanye nokuhlushwa okuqondiwe ukuze kuthuthukiswe ukusebenza futhi kuncishiswe ukungcolisa.
Ukucindezela kwe-Transmembrane (TMP) ku-Ultrafiltration
3.1. Kuyini Ukucindezeleka Kwe-Transmembrane?
Ingcindezi ye-Transmembrane (TMP) umehluko wengcindezi odlula i-membrane yokuhlunga, oqhuba i-solvent kusukela ohlangothini lokuphakelayo iye ohlangothini olungenamanzi. I-TMP ingamandla amakhulu ngemuva kwenqubo yokuhlukanisa ekuhlungeni kwe-ultrafiltration, okuvumela i-solvent ukuthi idlule ku-membrane ngenkathi igcina amaprotheni namanye ama-macromolecule.
Ifomula ye-TMP:
- Umehluko olula: TMP = P_feed − P_permeate
- Indlela yobunjiniyela: TMP = [(P_feed + P_retentate)/2] − P_permeate
Lapha, i-P_feed iyingcindezi yokungena, i-P_retentate iyingcindezi yokukhipha ohlangothini lwe-retentate, kanti i-P_permeate iyingcindezi yohlangothi lwe-permeate. Ukufaka ingcindezi ye-retentate (noma i-concentrate) kunikeza inani elinembile kakhulu ebusweni be-membrane, okubalwa ukuthambekela kwengcindezi okubangelwa ukumelana nokugeleza kanye nokungcola. - Ingcindezi yokudla kanye nesilinganiso sokugeleza
- Ingcindezi yokuqina (uma kusebenza)
- Ingcindezi egcwala (ngokuvamile enomoya)
- Ukumelana nolwelwesi
I-TMP iyahlukahluka ngohlobo lwe-membrane, ukwakheka kwesistimu, kanye nezimo zenqubo.
Ukulawula Iziguquguquko:
3.2. I-TMP kanye neNqubo ye-Ultrafiltration
I-TMP idlala indima ebalulekile ekuhlungeni kwamaprotheni e-ultrafiltration, iqhuba izixazululo zamaprotheni nge-membrane. Ingcindezi kumele ibe phezulu ngokwanele ukuze inqobe ukumelana okuvela ku-membrane nanoma yiziphi izinto eziqongelelwe kodwa ingabi phezulu kakhulu kangangokuthi isheshise ukungcola.
Ithonya Lokuqina Kwesisombululo Nokuhlushwa Kwamaprotheni
- Ukuqina kwezixazululo zamaprotheni:Ukuchama okuphezulu kwandisa ukumelana nokugeleza, okudinga i-TMP ephezulu ukuze kugcinwe ukuchama okufanayo. Isibonelo, ukwengeza i-glycerol ekudleni noma ukusebenza ngamaprotheni agxilile kukhulisa ukuchama futhi ngaleyo ndlela i-TMP esebenzayo iyadingeka.
- Ukuhlushwa kwamaprotheni:Njengoba ukuhlushwa kukhuphuka ngesikhathi sesigaba sokuhlushwa kwe-ultrafiltration, i-viscosity yesisombululo iyanda, i-TMP iyanda, futhi ingozi yokungcoliswa kwe-membrane noma i-concentration polarization iyanda.
- Umthetho kaDarcy:I-TMP, i-permeate flux (J), kanye ne-viscosity (μ) kuhlobene nge-TMP = J × μ × R_m (ukumelana nolwelwesi). Ukuze uthole izixazululo zamaprotheni anokugcwala okuphezulu, ukulungiswa kwe-TMP okucophelelayo kubalulekile ukuze kuhlungwe kahle.
Izibonelo:
- Ukuhlunga nge-ultrafiltration kwezixazululo ze-antibody eziqinile kudinga ukuphathwa kwe-TMP ngokucophelela ukuze kulwiswane nokwanda kwe-viscosity.
- I-PEGylation noma ezinye izinguquko zephrotheni zishintsha ukusebenzisana ne-membrane, okuthinta i-TMP edingekayo ukuze kuqhume okufunwayo.
3.3. Ukuqapha Nokuthuthukisa i-TMP
Ukugcina i-TMP ngaphakathiububanzi obujwayelekile bokucindezela kwe-transmembranekubalulekile ekusebenzeni kahle kwe-membrane ye-ultrafiltration eqinile kanye nekhwalithi yomkhiqizo. Ngokuhamba kwesikhathi, njengoba i-ultrafiltration iqhubeka, ukuhlukaniswa kwe-concentration kanye ne-fouling kungabangela i-TMP ukuthi ikhuphuke, ngezinye izikhathi ngokushesha.
Imikhuba Yokuqapha:
- Ukuqapha kwesikhathi sangempela:I-TMP ilandelwa ngokungena, ukubuyisa, kanye nokungena ngaphakathiama-pressure transmitter.
- I-Raman Spectroscopy:Isetshenziselwa ukuqapha okungangenisi ukungcola kwamaprotheni kanye nokugcwala kwe-excipient, okusiza ukulawula i-TMP eguquguqukayo ngesikhathi sokuhluzwa kwe-ultrafiltration kanye nokuhluzwa kwe-diafiltration.
- Ukulawula okuthuthukisiwe:Izihlungi ze-Kalman Extended (EKF) zingacubungula idatha yesensa, zilungise ngokuzenzakalelayo i-TMP ukuze kugwenywe ukungcoliswa ngokweqile.
- Setha i-TMP yokuqala ngaphakathi kobubanzi obujwayelekile:Akuphansi kakhulu ukunciphisa ukugeleza kwamanzi, akuphezulu kakhulu ukugwema ukungcola okusheshayo.
- Lungisa i-TMP njengoba i-viscosity ikhuphuka:Ngesikhathi sesigaba sokuhlushwa kwe-ultrafiltration, phakamisa i-TMP kancane kancane kuphela uma kudingeka.
- Lawula ukugeleza kokudla kanye ne-pH:Ukwandisa ukugeleza kokudla noma ukwehlisa i-TMP kunciphisa ukuhlukaniswa kokugxila kanye nokungcola.
- Ukuhlanzwa nokufakwa kwe-membrane:Ama-TMP aphezulu ahlotshaniswa nokuhlanzwa okuvamile kanye nokuncipha kwesikhathi sokuphila kwe-membrane.
Ukuthuthukisa Amasu:
Izibonelo:
- Ukungcola kokugqwala emigqeni yokucubungula amaprotheni kuholela ekwandeni kwe-TMP kanye nokuncipha kokuguquguquka, okudinga ukuhlanzwa noma ukushintshwa kwe-membrane ukuze kubuyiselwe ukusebenza okuvamile.
- Ukwelashwa kwangaphambili kwe-enzyme (isb., ukwengeza i-pectinase) kunganciphisa i-TMP futhi kwandise isikhathi sokuphila kwe-membrane ngesikhathi sokuhluzwa kwe-protein ye-rapeseed ene-viscosity ephezulu.
3.4. I-TMP ezinhlelweni ze-TFF
Ukuhlunga ukugeleza kwe-Tangential (transverse) (TFF) kusebenza ngokuhambisa isixazululo sokuphakelayo ngaphesheya kwe-membrane kunokuba sidlule kuyo ngqo, okuthonya kakhulu amandla e-TMP.
Ukulawulwa kanye nebhalansi ye-TMP
- Ingcindezi ye-transmembrane ye-TFF (i-TFF TMP):Ilawulwa ngokulawula kokubili izinga lokugeleza kokudla kanye nokucindezela kwephampu ukuze kugwenywe i-TMP eyeqile ngenkathi kwandiswa ukugeleza kwamanzi.
- Ukuthuthukisa amapharamitha:Ukwanda kokugeleza kokudla kunciphisa ukufakwa kwamaprotheni endaweni, kuzinzise i-TMP, futhi kunciphisa ukungcola kwe-membrane.
- Ukumodela kwekhompyutha:Amamodeli e-CFD abikezela futhi athuthukise i-TFF TMP ukuze kutholakale umkhiqizo, ubumsulwa, kanye nokukhiqiza okuphezulu kakhulu—okubaluleke kakhulu ezinqubweni ezifana ne-mRNA noma i-extracellular vesicle isolation.
Izibonelo:
- Ekucubungulweni kwezinto eziphilayo, i-TFF TMP engcono kakhulu iveza ukubuyiselwa kwe-mRNA okungaphezulu kuka-70% ngaphandle kokuwohloka, okudlula izindlela ze-ultracentrifugation.
- Ukulawulwa kwe-TMP okuguquguqukayo, okusekelwe kumamodeli ezibalo kanye nempendulo yezinzwa, kunciphisa imvamisa yokufakwa kwe-membrane futhi kuthuthukisa isikhathi sokuphila kwe-membrane ngokunciphisa ukungcoliswa.
Izinto ezibalulekile okufanele uzicabangele:
- Ingcindezi ye-TMP transmembrane kumele ilawulwe ngenkuthalo ku-TFF ukuze kulondolozwe ukusebenza kahle kwenqubo, ukuguquguquka, kanye nempilo ye-membrane.
- Ukulungiswa kwe-TMP okuhlelekile kunciphisa izindleko zokusebenza, kusekela ukubuyiswa komkhiqizo ohlanzekile kakhulu, futhi kwandise isikhathi sokuphila kwe-membrane ekuhlungeni amaprotheni kanye nezinqubo ezihlobene.
Izindlela Zokungcolisa Nobuhlobo Bazo Nokuqina
Izindlela Eziyinhloko Zokungcola Ekuhluzeni Amaphrotheni Nge-Ultrafiltration
Ukuhlunga amaprotheni ngokweqile kuthinteka yizindlela eziningana ezihlukene zokungcola:
Ukungcola Kokugqwala:Kwenzeka lapho imikhiqizo yokugqwala—ngokuvamile ama-iron oxides—iqoqana ezindaweni ze-membrane. Lokhu kunciphisa ukugeleza futhi kunzima ukususa ngezinto zokuhlanza zamakhemikhali ezijwayelekile. Ukungcola kokugqwala kuholela ekulahlekelweni okuqhubekayo kokusebenza kwe-membrane futhi kwandisa imvamisa yokufakwa kwe-membrane ngokuhamba kwesikhathi. Umthelela wayo unzima kakhulu nge-PVDF kanye ne-PES membranes ezisetshenziswa ekuphatheni amanzi kanye nokusetshenziswa kwamaprotheni.
Ukungcola Kwemvelo:Ibangelwa kakhulu amaprotheni afana ne-bovine serum albumin (BSA), futhi ingaqiniswa lapho kukhona ezinye izinto eziphilayo ezifana nama-polysaccharides (isb., i-sodium alginate). Izindlela zifaka phakathi ukumuncwa ema-pores e-membrane, ukuvalwa kwama-pore, kanye nokwakheka kwesendlalelo sekhekhe. Imiphumela ehambisanayo yenzeka lapho kukhona izingxenye eziningi eziphilayo, lapho izinhlelo ezixubile-foulant zibhekana nokungcola okukhulu kunokudla kweprotheni eyodwa.
Ukwehlukaniswa kokugxila:Njengoba ukuhlunga okungaphezulu kuqhubeka, amaprotheni agciniwe aqoqana eduze kobuso be-membrane, okwandisa ukuhlushwa kwendawo kanye ne-viscosity. Lokhu kudala ungqimba lwe-polarisation oluthuthukisa ukuthambekela kokungcola futhi lunciphise ukugeleza. Inqubo iyashesha njengoba isigaba sokuhlunga okungaphezulu sithuthuka, sithonywa ngqo yingcindezi ye-transmembrane kanye ne-flow dynamics.
Ukungcola kwe-Colloidal kanye ne-Mixed-Foulant:Izinto ezihlanganisiwe (isb., i-silica, amaminerali angaphili) zingase zihlangane namaprotheni, zidale izendlalelo ezihlanganisiwe eziyinkimbinkimbi ezenza kube nzima ukungcola kwe-membrane. Ukuba khona kwe-silica ye-colloidal, isibonelo, kunciphisa kakhulu amazinga okushintshashintsha, ikakhulukazi uma kuhlanganiswa nezinto eziphilayo noma ngaphansi kwezimo ze-pH ezingaphelele.
Ithonya Lokuqina Kwesixazululo Ekuthuthukisweni Kokungcola
Ukuqina kwezixazululo zamaprotheni kuthinta kakhulu i-kinetics ye-fouling kanye nokuhlanganiswa kwe-membrane:
Ukungcola Okusheshisiwe:Ukuqina okuphezulu kwesisombululo samaprotheni kwandisa ukumelana nokuthuthwa kwe-solutes ezigciniwe emuva, okwenza kube lula ukwakheka kwengqimba yekhekhe ngokushesha. Lokhu kukhulisa ingcindezi ye-transmembrane (TMP), kusheshise ukuqina kwe-membrane kanye nokungcola.
Imiphumela Yokwakheka Kwesixazululo:Uhlobo lwephrotheni lushintsha i-viscosity; amaprotheni e-globular (isb., i-BSA) kanye namaprotheni anwetshiwe aziphatha ngendlela ehlukile ngokuphathelene nokugeleza kanye nokwahlukana. Ukwengeza ama-compounds afana nama-polysaccharides noma i-glycerol kukhulisa kakhulu i-viscosity, okukhuthaza ukungcola. Izithasiselo kanye nokuhlanganiswa kwamaprotheni ekugxilweni okuphezulu kuqinisa kakhulu izinga lapho ama-membrane evaleka khona, okunciphisa ngqo kokubili ukugeleza kanye nokuphila kwe-membrane.
Imiphumela Yokusebenza:Ukuchayeka okuphezulu kudinga i-TMP eyengeziwe ukuze kugcinwe amazinga okuhlunga ezinqubweni zokuhlunga ezihambayo. Ukuchayeka isikhathi eside ku-TMP ephezulu kukhulisa ukungcola okungenakuguqulwa, okuvame ukudinga ukuhlanzwa kwe-membrane njalo noma ukushintshwa kwe-membrane kusenesikhathi.
Indima Yezimpawu Zokuphakelayo
Izici zokudla—okungukuthi izakhiwo zamaprotheni kanye nekhemikhali yamanzi—zinquma ubukhali bokungcola:
Usayizi Wamaprotheni Nokusatshalaliswa Kwawo:Amaprotheni amakhulu noma ahlanganisiwe anomkhuba omkhulu wokubangela ukuvimba kwe-pore kanye nokwakheka kwamakhekhe, okuphakamisa ukuthambekela kokunamathelana kanye nokuminyana ngesikhathi sokuhlushwa kwamaprotheni okuhlushwa kakhulu.
i-pH:I-pH ephakeme ikhulisa ukuxoshwa kwe-electrostatic, ivimbela amaprotheni ukuthi ahlangane eduze kwe-membrane, ngaleyo ndlela inciphise ukungcoliswa. Ngokuphambene nalokho, izimo ze-acidic zinciphisa ukuxoshwa, ikakhulukazi kwi-colloidal silica, okwenza ukungcoliswa kwe-membrane kube kubi kakhulu futhi kwehlise amazinga okugeleza.
Izinga lokushisa:Amazinga okushisa aphansi enqubo ngokuvamile anciphisa amandla e-kinetic, okunganciphisa amazinga okungcolisa kodwa futhi kwandise ukuchachamba kwesisombululo. Amazinga okushisa aphezulu asheshisa ukungcolisa kodwa futhi angathuthukisa ukusebenza kahle kokuhlanza.
Izinto Ezingaphilile/Ezingavundile:Ukuba khona kwe-colloidal silica noma izinsimbi kwandisa ukungcola, ikakhulukazi ngaphansi kwezimo ze-asidi. Izinhlayiya ze-silica zandisa ukujiya kwesisombululo sonke futhi zivimbele izikhala ngokomzimba, okwenza ukuhlushwa kwe-ultrafiltration kungasebenzi kahle futhi kunciphisa isikhathi sokuphila kwe-membrane kanye nokusebenza kwayo.
Ukwakheka kwe-Ionic:Ukwengeza izinhlobo ezithile ze-ionic (Na⁺, Zn²⁺, K⁺) kunganciphisa ukungcola ngokushintsha amandla kagesi kanye nokumanzisa phakathi kwamaprotheni nama-membrane. Kodwa-ke, ama-ion afana ne-Ca²⁺ avame ukukhuthaza ukuhlangana futhi andise amandla okungcola.
Izibonelo:
- Ngesikhathi sokuhlunga ukugeleza okuphambene, ukudla okucebile ngamaprotheni anesisindo sama-molecule aphezulu kanye nokuthambekela okuphezulu kuzobhekana nokwehla okusheshayo kokuguquguquka, okwandisa ukuhlanza kanye nemikhuba yokushintsha.
- Uma amanzi okuphakelayo equkethe i-colloidal silica futhi eba ne-asidi, ukuhlanganiswa kwe-silica kanye nokufakwa kwayo kuyanda, okwandisa kakhulu amazinga okungcolisa kanye nokusebenza kwe-membrane kwehla.
Ngamafuphi, ukuqonda ukusebenzisana phakathi kwe-viscosity yesisombululo, izinhlobo zokungcolisa, kanye nezici zokudla kubalulekile ekwenzeni ngcono ukuhlushwa kwe-ultrafiltration, ukunciphisa ukungcolisa kwe-membrane, kanye nokwandisa isikhathi sokuphila kwe-membrane.
Ukuhlukaniswa Kokuhlushwa Nokuphathwa Kwakho
Kuyini ukuhlukaniswa kokugxila?
I-Concentration polarization ukuqongelela kwendawo kwe-solute egciniwe—njengamaprotheni—esixhumi se-membrane/solution ngesikhathi sokuhluzwa kwe-ultrafiltration. Uma kukhulunywa ngezixazululo zamaprotheni, njengoba uketshezi lugeleza ngokumelene ne-membrane engena kancane, amaprotheni anqatshwe yi-membrane avame ukunqwabelana ngengqimba encane yomngcele eduze kobuso. Lokhu kwakheka kuphumela ekujuleni okukhulu kokuhlushwa: ukuhlushwa okuphezulu kwamaprotheni khona kanye ku-membrane, okuphansi kakhulu kusisombululo esikhulu. Lesi simo siyaguqulwa futhi silawulwa amandla e-hydrodynamic. Sihlukile ekungcoleni kwe-membrane, okubandakanya ukufakwa okungapheli noma ukumuncwa ngaphakathi noma ku-membrane.
Indlela Ukuhlukaniswa Kokuhlushwa Okukhuphula Ngayo Ukuqina Nokungcola
Ebusweni be-membrane, ukuqongelela okuqhubekayo kwamaprotheni kwakha ungqimba lomngcele okwandisa ukuhlushwa kwe-solute yendawo. Lokhu kunemiphumela emibili ebalulekile:
Ukwanda Kokugoba Kwendawo:Njengoba ukugcwala kwamaprotheni kukhuphuka eduze kwe-membrane, ukugcwala kwesisombululo samaprotheni kule ndawo encane nakho kuyanda. Ukugcwala okuphezulu kuvimbela ukuthuthwa kwe-solute emuva kwe-membrane, kuqinise kakhulu i-gradient yokugcwala futhi kudale i-feedback loop yokumelana nokwanda kokugeleza. Lokhu kuphumela ekunciphiseni ukugeleza kwe-permeate kanye nesidingo samandla aphezulu sokuqhubeka nokuhlunga.
Ukusiza Ukungcola Kwe-Membrane:Ukuhlushwa okuphezulu kwamaprotheni eduze kwe-membrane kwandisa amathuba okuhlangana kwamaprotheni, futhi kwezinye izinhlelo, ukwakheka kwesendlalelo sejeli. Lolu ngqimba luvimbela izikhala ze-membrane futhi lukhulisa ukumelana nokugeleza. Izimo ezinjalo zilungele ukuqala kokungcola okungenakuguqulwa, lapho ukuhlanganiswa kwamaprotheni kanye nokungcola kubopha ngokomzimba noma ngamakhemikhali ku-membrane matrix.
Ukuthwebula izithombe kokuhlola (isb., i-electron microscopy) kuqinisekisa ukuhlangana okusheshayo kwamaqoqo amaprotheni amancane kulwelwesi, okungakhula kube amadiphozithi abalulekile uma izilungiselelo zokusebenza zingaphathwa ngendlela efanele.
Amasu Okunciphisa Ukwehlukaniswa Kokugxila
Ukuphatha ukuhlukaniswa kwe-concentration ekuhlungeni kwamaprotheni okuhlungwa kwe-ultrafiltration noma ekuhlungeni ukugeleza okuguquguqukayo kudinga indlela ekabili: ukulungisa i-hydrodynamics kanye nokulungisa amapharamitha okusebenza.
Ukuthuthukiswa Kwejubane Elihambayo:
Ukuphakamisa ijubane lokugeleza okuwelayo kwandisa ukugeleza kwe-tangential kulo lonke ulwelwesi, kukhuthaza ukucheba nokunciphisa ungqimba lomngcele wokuhlushwa. Ukucheba okunamandla kakhulu kususa amaprotheni aqoqiwe ebusweni belwelwesi, kunciphisa kokubili ukuhlukaniswa kanye nengozi yokungcola. Isibonelo, ukusebenzisa ama-mixer aqinile noma ukufaka i-gas sparging kuphazamisa ungqimba oluncibilikayo, ikakhulukazi ukuthuthukisa ukugeleza kokungena kanye nokusebenza kahle enqubweni yokuhlunga ukugeleza okuwelayo.
Ukushintsha Amapharamitha Okusebenza:
Ukucindezela kwe-Transmembrane (TMP):I-TMP umehluko wokucindezela kulo lonke ulwelwesi kanye namandla aqhubayo okuhluza nge-ultrafiltration. Kodwa-ke, ukusunduza i-TMP phezulu ukuze kusheshiswe ukuhlunga kungabangela umonakalo ngokuqinisa ukuhlukaniswa kokugxila. Ukunamathela ebangeni elijwayelekile lokucindezela kwe-transmembrane—okungadluli imikhawulo ebekiwe yokuhluza nge-protein—kusiza ukuvimbela ukwakheka ngokweqile kwe-solute kanye nokwanda okuhlobene kwe-viscosity yendawo.
Izinga Lokugunda:Izinga lokucheba, umsebenzi wejubane lokuhamba kwe-cross-flow kanye nomklamo wesiteshi, lidlala indima ebalulekile ekushintsheni kokuthutha okune-solute. Ukucheba okuphezulu kugcina ungqimba lwe-polarization luncane futhi luhamba, okuvumela ukuvuselelwa njalo kwesifunda esinciphile esiseduze ne-membrane. Ukwandisa izinga lokucheba kunciphisa isikhathi amaprotheni anaso sokuqongelela futhi kunciphisa ukukhuphuka kwe-viscosity endaweni exhumanayo.
Izakhiwo Zokuphakelayo:Ukulungisa izakhiwo zesisombululo seprotheyini engenayo—njengokwehlisa i-viscosity yesisombululo seprotheyini, ukunciphisa okuqukethwe okuhlanganisiwe, noma ukulawula i-pH kanye namandla e-ionic—kungasiza ekunciphiseni ubukhulu kanye nomthelela wokuhlukaniswa kwe-concentration. Ukwelashwa kwangaphambili kokudla kanye nezinguquko zokwakheka kungathuthukisa ukusebenza kwe-membrane ye-ultrafiltration futhi kwandise isikhathi sokuphila kwe-membrane ngokunciphisa imvamisa yokuhlanza i-membrane.
Isibonelo Sokusetshenziswa:
Isitshalo esisebenzisa i-tangential flow filtration (TFF) ukugxila ama-antibodies e-monoclonal sisebenzisa ijubane elithuthukisiwe ngokucophelela lokugeleza futhi sigcina i-TMP ngaphakathi kwefasitela eliqinile. Ngokwenza kanjalo, abaqhubi banciphisa ukuhlukaniswa kwe-concentration kanye nokungcola kwe-membrane, banciphisa kokubili imvamisa yokufakwa kwe-membrane kanye nemijikelezo yokuhlanza—ukunciphisa ngqo izindleko zokusebenza nokuthuthukisa isivuno somkhiqizo.
Ukulungiswa okufanele kanye nokuqapha lezi ziguquguquko—kufaka phakathi ukulinganiswa kwe-viscosity yesisombululo seprotheyini ngesikhathi sangempela—kuyisisekelo sokwenza ngcono ukusebenza kokuhlushwa kwe-ultrafiltration kanye nokunciphisa imiphumela emibi ehlobene nokuhlukaniswa kokuhlushwa ekucutshungulweni kwamaprotheni.
Ukuthuthukisa i-Ultrafiltration yezixazululo ze-High-Vicosity Protein
6.1. Imikhuba Emihle Yokusebenza
Ukugcina ukusebenza kahle kokuhlunga kwe-ultrafiltration ngezixazululo zamaprotheni ane-viscosity ephezulu kudinga ibhalansi ebucayi phakathi kwengcindezi ye-transmembrane (TMP), ukuhlushwa kwamaprotheni, kanye ne-viscosity yesisombululo. I-TMP—umehluko ekucindezelweni kulo lonke ulwelwesi—ithonya ngqo izinga lokuhlushwa kwamaprotheni ane-ultrafiltration kanye nezinga lokungcoliswa kwe-membrane. Lapho kucutshungulwa izixazululo ezine-viscous ezifana nama-antibodies e-monoclonal noma amaprotheni e-serum ane-concentration ephezulu, noma yikuphi ukwanda okweqile kwe-TMP kungakhuphula ukugeleza ekuqaleni, kodwa futhi kusheshisa ngokushesha ukungcoliswa kanye nokuqongelela kwamaprotheni ebusweni be-membrane. Lokhu kuholela enkambisweni yokuhlunga ebucayi nengazinzile, eqinisekiswe yizifundo zezithombe ezibonisa izendlalelo zamaprotheni aminyene ezakha amazinga aphezulu e-TMP kanye namaprotheni angaphezu kuka-200 mg/mL.
Indlela engcono kakhulu ihilela ukusebenzisa uhlelo eduze, kodwa lungadluli, i-TMP ebalulekile. Kuleli qophelo, umkhiqizo ukhuliswa kakhulu kodwa ingozi yokungcola okungenakuguqulwa ihlala incane kakhulu. Ngokubukeka okuphezulu kakhulu, okutholakele kwakamuva kusikisela ukunciphisa i-TMP kanye nokwandisa ukugeleza kokudla ngasikhathi sinye (ukuhlunga ukugeleza okuguquguqukayo) ukusiza ekunciphiseni ukuhlukaniswa kokuhlushwa kanye nokufakwa kwamaprotheni. Isibonelo, izifundo zokuhlushwa kwamaprotheni e-Fc-fusion zibonisa ukuthi izilungiselelo ze-TMP eziphansi zisiza ukugcina ukugeleza okuzinzile ngenkathi kunciphisa ukulahleka komkhiqizo.
Ukwanda kancane kancane nangokuhlelekile kokuhlushwa kwamaprotheni ngesikhathi sokuhluzwa kwe-ultra-filtration kubalulekile. Izinyathelo zokuhlushwa ngokuzumayo zingaphoqa isisombululo ukuba singene ohlelweni olunamandla kakhulu ngokushesha, okwandisa izingozi zokuhlanganiswa kanye nobunzima bokungcola. Esikhundleni salokho, ukuphakamisa amazinga amaprotheni kancane kancane kuvumela ukuthi imingcele yenqubo efana ne-TMP, ijubane lokugeleza kwe-cross-flow, kanye ne-pH ilungiswe ngesikhathi esifanayo, okusiza ukugcina ukuzinza kwesistimu. Izifundo zamacala okuhlushwa kwe-enzyme ziqinisekisa ukuthi ukugcina izingcindezi zokusebenza eziphansi phakathi nalezi zigaba kuqinisekisa ukwanda okulawulwayo kokuhlushwa, kunciphisa ukwehla kokuguquguquka ngenkathi kuvikela ubuqotho bomkhiqizo.
6.2. Ukuvama Kokushintsha Ulwelwesi Nokunakekelwa
Ukuvama kokufakwa kwe-membrane ekuhlungeni kwe-ultrafiltration kuhlobene kakhulu nezinkomba zokungcola kanye nokwehla kokuguquguquka. Esikhundleni sokuthembela kuphela ekunciphiseni kokuguquguquka okuhlobene njengesibonakaliso sokuphela kokuphila, ukuqapha ukumelana kokungcola okuthile—isilinganiso sokulinganisa esimele ukumelana okubangelwa izinto eziqongelelwe—kuye kwabonakala kuthembekile kakhulu, ikakhulukazi ekudleni okuxubile kwamaprotheni noma kwamaprotheni-polysaccharide, lapho ukungcola kungenzeka khona ngokushesha nangokubi kakhulu.
Ukuqapha izinkomba ezengeziwe zokungcola nakho kubalulekile. Izimpawu ezibonakalayo zokubekwa kobuso, ukugeleza okungalingani kokungena, noma ukwanda okuqhubekayo kwe-TMP (naphezu kokuhlanza) konke kuyizibonakaliso zesixwayiso zokungcola okuthuthukile okungaphambi kokwehluleka kwe-membrane. Amasu anjengokulandelela i-modified fouling index (MFI-UF) nokuyihlobanisa nokusebenza kwe-membrane avumela ukuhlela okubikezelayo kokufakwa esikhundleni kunezinguquko ezisabelayo, ngaleyo ndlela kuncishiswe isikhathi sokungasebenzi kanye nokulawula izindleko zokulungisa.
Ubuqotho be-membrane abugcini nje ngokwakheka kwe-organic foulant kodwa futhi nokugqwala, ikakhulukazi ezinqubweni ezisebenza ku-pH eyeqile noma ezinosawoti omningi. Ukuhlolwa okuvamile kanye nemikhuba yokuhlanza amakhemikhali kufanele kuqalwe ukuze kulawulwe kokubili ukugqwala kanye nokufakwa kwe-foulant. Uma kuqashelwa ukungcoliswa okuhlobene nokugqwala, imvamisa yokuhlanza i-membrane kanye nezikhawu zokufaka esikhundleni kumele kulungiswe ukuqinisekisa ukuthi i-membrane ihlala isikhathi eside kanye nokusebenza kwe-membrane okuqhubekayo. Ukugcinwa okuphelele nokuhleliwe kubalulekile ekunciphiseni umthelela walezi zinkinga kanye nokwandisa ukusebenza okuphumelelayo.
6.3. Ukulawulwa Kwenqubo kanye Nokulinganiswa Kokugoba Okungaphakathi Komugqa
Ukulinganisa okunembile, kwesikhathi sangempela kwe-viscosity yesisombululo seprotheyini kubalulekile ekulawuleni inqubo ekuhlungeni kwe-ultrafiltration, ikakhulukazi njengoba ukugxila kanye ne-viscosities kukhuphuka. Izinhlelo zokulinganisa i-viscosity eziqondile zinikeza ukuqapha okuqhubekayo, okuvumela impendulo esheshayo futhi kuvumela ukulungiswa okuguquguqukayo kumapharamitha wesistimu.
Ubuchwepheshe obusha bushintshe isimo sokulinganisa i-viscosity yesisombululo samaprotheni:
I-Raman Spectroscopy ene-Kalman Filtering: Ukuhlaziywa kweRaman kwesikhathi sangempela, okusekelwa yizihlungi zeKalman ezinwetshiwe, kwenza kube lula ukulandelela okuqinile kokuhlushwa kwamaprotheni kanye nokwakheka kwe-buffer. Le ndlela yandisa ukuzwela nokunemba, isekela ukuzenzekela kwenqubo yokugxila kokuhlushwa kwe-ultrafiltration kanye nokuhlushwa kwe-diafiltration.
I-Kinematic Capillary Viscometry Ezenzakalelayo: Isebenzisa umbono wekhompyutha, lobu buchwepheshe bulinganisa ngokuzenzakalelayo i-viscosity yesisombululo, bunqoba amaphutha enziwe ngesandla futhi bunikeza ukuqapha okuphindaphindwayo, okuphindaphindwayo emifudlaneni eminingi yenqubo. Buqinisekiswa kokubili ukwakheka kwamaprotheni okujwayelekile nokuyinkimbinkimbi futhi kunciphisa ukungenelela ngesikhathi sesigaba sokuhlushwa kwe-ultrafiltration.
Amadivayisi e-Microfluidic Rheology: Izinhlelo ze-Microfluidic zinikeza amaphrofayili e-rheological anemininingwane, aqhubekayo, ngisho nasezixazululweni zamaprotheni ezingezona eze-Newtonian, ezinamaprotheni amaningi. Lezi ziwusizo kakhulu ekukhiqizweni kwemithi, ukusekela amasu obuchwepheshe bokuhlaziya inqubo (i-PAT) kanye nokuhlanganiswa nezihibe zempendulo.
Ukulawulwa kwenqubo kusetshenziswa la mathuluzi kwenza kube lula ukusebenzisa ama-feedback loops okulungisa i-TMP ngesikhathi sangempela, izinga lokuphakelayo, noma ijubane lokuhamba ngokuphambuka ekuphenduleni izinguquko ze-viscosity. Isibonelo, uma ukuzwa okuqondile kuthola ukwanda okungazelelwe kwe-viscosity (ngenxa yokwanda kokuhlushwa noma ukuhlanganiswa), i-TMP ingancishiswa ngokuzenzakalelayo noma ijubane lokuhamba ngokuphambuka liphakanyiswe ukuze kukhawulelwe ukuqala kwe-concentration polarization ekuhlungeni okungaphezulu. Le ndlela ayigcini nje ngokwandisa isikhathi sokuphila kwe-membrane kodwa futhi isekela ikhwalithi yomkhiqizo ehambisanayo ngokuphatha izici ezithinta i-viscosity yezixazululo zamaprotheni ngokuguquguqukayo.
Ukukhethwa kobuchwepheshe bokuqapha i-viscosity obufanele kakhulu kuncike ezidingweni ezithile zohlelo lokusebenza lwe-ultrafiltration, kufaka phakathi ububanzi be-viscosity obulindelekile, ubunzima bokwakheka kwamaprotheni, izidingo zokuhlanganisa, kanye nezindleko. Lokhu kuthuthuka ekuqapheni ngesikhathi sangempela kanye nokulawula inqubo enamandla kuthuthukise kakhulu ikhono lokwenza ngcono i-ultrafiltration yezixazululo zamaprotheni ane-viscosity ephezulu, okuqinisekisa kokubili ukuzinza kokusebenza kanye nokukhiqizwa okuphezulu komkhiqizo.
Ukuxazulula Izinkinga Nezinkinga Ezivamile Ekuhluzeni Amaphrotheni Nge-Ultrafiltration
7.1. Izimpawu, Izimbangela, kanye Nezindlela Zokwelapha
Ukucindezela Okukhuphukile Kwe-Transmembrane
Ukwenyuka kwengcindezi ye-transmembrane (TMP) ngesikhathi sokuhluzwa kwe-ultra-filtration kubonisa ukumelana okukhula kulo lonke ulwelwesi. Imiphumela yengcindezi ye-transmembrane ekuhluzweni kwe-ultra-filtration iqondile: ububanzi obujwayelekile bengcindezi ye-transmembrane buvame ukuxhomeka enqubweni, kodwa ukwanda okuqhubekayo kufanele kuhlolwe. Izimbangela ezimbili ezivamile ziyavela:
- I-viscosity ephezulu yesisombululo seprotheni:Njengoba i-viscosity yezixazululo zamaprotheni ikhula—ngokuvamile lapho kuhlungwa kakhulu amaprotheni e-ultrafiltration—ingcindezi edingekayo ukuze kugeleze iyanda. Lokhu kuzwakala ezinyathelweni zokugcina zokuhlungwa kanye ne-diafiltration lapho izixazululo zihlungwa kakhulu.
- Ukungcola kwe-membrane:Izinto ezibangela ukubola ezifana nama-protein aggregates noma izingxube ze-polysaccharide-protein zinganamathela noma zivale ama-membrane pores, okuholela ekukhuphukeni okusheshayo kwe-TMP.
Amakhambi:
- Yehlisa i-TMP futhi wandise ukugeleza kokudla: Ukunciphisa i-TMP ngenkathi kukhulisa ijubane lokuphakelwa kunciphisa ukuhlukaniswa kokugxila kanye nokwakheka kwezingqimba zejeli, okukhuthaza ukugeleza okuzinzile.
- Ukuhlanza njalo ulwelwesi: Misa imvamisa yokuhlanza i-membrane efanele ukuze ususe ama-foulants aqongelelekile. Qapha ukusebenza kahle ngokulinganisa i-viscosity yesisombululo seprotheni ngemva kokuhlanza.
- Buyisela ulwelwesi olugugayo: Ukwanda kwemvamisa yokufakwa kwe-membrane kungadingeka uma ukuhlanzwa kunganele noma isikhathi sokuphila kwe-membrane sifinyelelwe.
Izinga Lokwehla Kokuguquguquka: Isihlahla Sokuxilonga
Ukwehla okuqhubekayo kokuguquguquka ngesikhathi sesigaba sokuhlushwa kwe-ultrafiltration kubonisa ukukhathazeka ngokukhiqiza. Landela le ndlela yokuxilonga:
- Gada i-TMP kanye ne-viscosity:Uma kokubili kukhuphukile, hlola ukuthi kukhona yini ukungcola noma ungqimba lwejeli.
- Hlola ukwakheka kokudla kanye ne-pH:Ukushintsha lapha kungashintsha ukujiya kwezixazululo zamaprotheni futhi kukhuthaze ukungcoliswa.
- Hlola ukusebenza kwe-membrane:Ukuncipha kokuphuma kwamanzi naphezu kokuhlanza izimpawu zokulimala kwe-membrane noma ukungcola okungenakuguqulwa.
Izixazululo:
- Lungiselela izinga lokushisa, i-pH, kanye namandla e-ionic ekudleni ukuze unciphise ukungcoliswa kanye nokuhlukaniswa kokugxila ekuhluzweni kwe-ultrafiltration.
- Sebenzisa amamojula e-membrane aguquliwe noma ajikelezayo ukuze uphazamise izendlalelo zejeli futhi ubuyisele ukugeleza.
- Yenza njalo ukulinganisa i-viscosity yesisombululo seprotheni ukuze ubikezele izinguquko ezithinta ukugeleza.
Ukwakheka Kwezingqimba Ezisheshayo Zokungcola Noma Zejeli
Ukwakheka kwengqimba yejeli okusheshayo kubangelwa ukuhlukaniswa okukhulu kokugxila ebusweni be-membrane. Ingcindezi ye-transverse flow filtration (TFF) transmembrane isengozini ikakhulukazi ngaphansi kwezimo zokudla ezine-viscosity ephezulu noma ezinamaphrotheni amaningi.
Amasu Okunciphisa Ubunzima:
- Faka izindawo ze-membrane ezihlala emanzini, ezishajeke kabi (isb., i-Polyvinylidene fluoride [PVDF] membranes) ukuze unciphise ukubopha nokunamathela kweprotheni.
- Phakamisa ukudla kusengaphambili usebenzisa i-coagulation noma i-electrocoagulation ukuze ususe izinto ezingcolisa kakhulu ngaphambi kokuhluzwa kakhulu.
- Hlanganisa amadivayisi emishini afana namamojula ajikelezayo enqubweni yokuhlunga ukugeleza okuphambene ukuze unciphise ukujiya kwesendlalelo sekhekhe futhi ulibazise ukwakheka kwesendlalelo sejeli.
7.2. Ukulungisa Ukuguquguquka Kokuphakelayo
Izinhlelo zokuhlunga amaprotheni amaningi kumele zivumelane nokuguquguquka kwezakhiwo noma ukwakheka kwamaprotheni okuphakelayo. Izici ezithinta ukujiya kwezixazululo zamaprotheni—njengokwakheka kwe-buffer, ukuhlushwa kwamaprotheni, kanye nokuthambekela kokuhlanganisa—zingashintsha ukuziphatha kwesistimu.
Amasu Okuphendula
- Ukuqapha ukuchayeka kanye nokwakheka kwesikhathi sangempela:Sebenzisa izinzwa zokuhlaziya eziku-inthanethi (i-Raman spectroscopy + i-Kalman filtering) ukuze kutholakale ngokushesha izinguquko zokuphakelayo, izindlela ze-UV noma ze-IR ezisebenza kahle kakhulu.
- Ukulawulwa kwenqubo yokuzivumelanisa nezimo:Lungisa izilungiselelo zepharamitha (izinga lokugeleza, i-TMP, ukukhethwa kwe-membrane) ekuphenduleni izinguquko ezitholakele. Isibonelo, ukwanda kwe-viscosity yesisombululo seprotheni kungadinga i-TMP ephansi kanye namazinga aphezulu okucheba.
- Ukukhetha ulwelwesi:Sebenzisa ama-membrane anosayizi we-pore kanye ne-chemistry engaphezulu elungiselelwe izakhiwo zokudla zamanje, ukulinganisela ukugcinwa kwamaprotheni kanye nokuguquguquka kwawo.
- Ukwelashwa kwangaphambi kokudla:Uma ukushintsha okungazelelwe kwemvelo yokudla kukhuthaza ukungcola, faka izinyathelo zokugaya noma zokuhlunga phezulu kwe-ultrafiltration.
Izibonelo:
- Ekucutshungulweni kwezinto eziphilayo, ukushintsha kwe-buffer noma izinguquko kuma-antibody aggregates kufanele kubangele ukulungiswa kwe-TMP kanye nokugeleza ngokusebenzisa uhlelo lokulawula.
- Ngokuhlungwa kwe-ultra-linked chromatography, ama-algorithms optimization adaptive mixing-integer optimization anganciphisa ukuguquguquka futhi anciphise izindleko zokusebenza ngenkathi egcina ukusebenza kwe-ultrafiltration membrane.
Ukulandelela njalo ukulinganiswa kwe-viscosity yesisombululo seprotheni kanye nokulungiswa okusheshayo kwezimo zenqubo kusiza ukuthuthukisa ukuhlushwa kwe-ultrafiltration, ukugcina ukuphuma, kanye nokunciphisa ukungcoliswa kwe-membrane kanye ne-concentration polarization.
imibuzo ejwayelekile ukubuzwa
8.1. Liyini izinga elijwayelekile lokucindezela kwe-transmembrane ekuhluzweni kwezixazululo zamaprotheni nge-ultrafiltration?
Ububanzi obujwayelekile bokucindezela kwe-transmembrane (TMP) ezinhlelweni zokuhlushwa kwamaprotheni e-ultrafiltration buxhomeke ohlotsheni lwe-membrane, ukwakheka kwemojuli, kanye nezici zokudla. Ezinqubweni eziningi ze-protein ultrafiltration, i-TMP ivame ukugcinwa phakathi kwebha eli-1 kuya kwele-3 (15–45 psi). Amanani e-TMP angaphezu kwe-0.2 MPa (cishe ama-29 psi) angabeka engcupheni umonakalo we-membrane, ukungcola okusheshayo, kanye nokuphila kwe-membrane okufushane. Ezisetshenzisweni ze-biomedical kanye ne-bioprocessing, i-TMP enconyiwe ngokuvamile akufanele idlule ibha eli-0.8 (~12 psi) ukuze kugwenywe ukuqhekeka kwe-membrane. Ngezinqubo ezifana nokuhlunga okuhambayo, ukuhlala ngaphakathi kwalolu hlu lwe-TMP kuvikela kokubili ukukhiqizwa kanye nobuqotho be-protein.
8.2. Ukujiya kwezixazululo zamaprotheni kuthinta kanjani ukusebenza kwe-ultrafiltration?
Ukuqina kwesisombululo samaprotheni kuthinta ngqo ukusebenza kokuhlushwa kwe-ultrafiltration. Ukuqina okuphezulu kwandisa ukumelana kokugeleza futhi kuphakamisa i-TMP, okuholela ekunciphiseni ukugeleza kwe-permeating kanye nokungcoliswa kwe-membrane okusheshayo. Lo mphumela ubonakala ngama-antibodies e-monoclonal noma amaprotheni e-Fc-fusion ekuhlushweni okuphezulu, lapho i-viscosity ikhula ngenxa yokusebenzisana kwamaprotheni namaphrotheni kanye nemiphumela yokushaja. Ukuphatha nokwenza ngcono i-viscosity ngama-excipients noma ukwelashwa kwe-enzyme kuthuthukisa ukugeleza, kunciphisa ukungcoliswa, futhi kuvumela ukuhlushwa okuphezulu okufinyelelekayo ngesikhathi sesigaba sokuhlushwa kwe-ultrafiltration. Ukuqapha ukulinganiswa kwe-viscosity yesisombululo samaprotheni kubalulekile ekugcineni ukucubungula okusebenzayo.
8.3. Kuyini ukuhlukaniswa kwe-concentration futhi kungani kubalulekile ku-TFF?
Ukuhlukaniswa kokuhlushwa ...
8.4. Nginquma kanjani ukuthi ngizoyishintsha nini i-membrane yami ye-ultrafiltration?
Shintsha i-membrane ye-ultrafiltration uma ubona ukwehla okuphawulekayo kokuphuma (ukuguquguquka), ukwanda okuqhubekayo kwe-TMP okungenakuxazululwa ukuhlanza okujwayelekile, noma ukungcola okubonakalayo okusalayo ngemva kokuhlanza. Izinkomba ezengeziwe zifaka phakathi ukulahlekelwa ukukhetha (ukwehluleka ukwenqaba amaprotheni aqondiwe njengoba kulindelekile) kanye nokungakwazi ukufinyelela ezicacisweni zokusebenza. Ukuqapha imvamisa yokufakwa kwe-membrane ngokuhlolwa okuvamile kokushintsha kanye nokukhetha kuyisisekelo sokukhulisa isikhathi sokuphila kwe-membrane ezinqubweni zokuhlushwa kwe-protein solution ultrafiltration.
8.5. Yimaphi amapharamitha okusebenza engingawalungisa ukuze nginciphise ukungcoliswa kwamaprotheni ku-TFF?
Amapharamitha abalulekile okusebenza ukunciphisa ukungcola kwamaprotheni ekuhlungeni ukugeleza okuphambeneyo afaka:
- Gcina ijubane elanele lokuhamba kwe-cross ukuze unciphise ukwakheka kwamaprotheni endawo futhi ulawule ukuhlukaniswa kokugxila.
- Sebenzisa ngaphakathi kobubanzi be-TMP obunconywayo, ngokuvamile obuyi-3–5 psi (0.2–0.35 bar), ukuvimbela ukuvuza okweqile komkhiqizo kanye nomonakalo we-membrane.
- Sebenzisa izindlela zokuhlanza ulwelwesi njalo ukuze unciphise ukungcola okungenakuguqulwa.
- Gada futhi, uma kudingeka, lungisa isisombululo sokudla kusengaphambili ukuze ulawule i-viscosity (isibonelo, usebenzisa ukwelashwa nge-enzyme njenge-pectinase).
- Khetha izinto ze-membrane kanye nosayizi we-pore (MWCO) ezifanele usayizi weprotheyini eqondiwe kanye nemigomo yenqubo.
Ukuhlanganisa i-hydrocyclone prefiltration noma i-enzymatic pretreatment kungathuthukisa ukusebenza kwesistimu, ikakhulukazi kuma-feed ane-viscosity ephezulu. Landela eduze ukwakheka kokudla bese ulungisa izilungiselelo ngendlela eguquguqukayo ukuze unciphise ukungcola kwe-membrane futhi uthuthukise isigaba sokuhlushwa kwe-ultrafiltration.
Isikhathi sokuthunyelwe: Novemba-03-2025
