Kugenzura Viscosity ya Poroteyine mu Gutunganya Utunyangingo (Ultrafiltration)

Gukoresha uburyo bwa Ultrafiltration kugira ngo habeho ibisubizo bya poroteyine nyinshi

6.1. Uburyo bwiza bwo gukoresha

Kugumana imikorere myiza yo kuyungurura ingano y’amaraso hakoreshejwe poroteyine ifite ubukana bwinshi bisaba uburinganire bwiza hagati y’umuvuduko wa transmembrane (TMP), ingano ya poroteyine, n’ubukana bw’umuti. TMP—itandukaniro ry’umuvuduko ku ruhu—bigira ingaruka ku muvuduko wa poroteyine ifite ubukana bwinshi n’urugero rw’ubukana bw’amaraso mu ruhu. Mu gutunganya imiterere y’amaraso nk’iy’umubiri urwanya uburwayi bwa monoclonal cyangwa poroteyine zifite ubukana bwinshi mu maraso, kwiyongera gukabije kwa TMP bishobora kongera ubwinshi bw’amaraso mu ntangiriro, ariko nanone bikongera vuba ubukana n’ubwiyongere bwa poroteyine ku buso bw’urukiramende. Ibi bituma inzira yo kuyungurura idakora neza kandi idahindagurika, byemejwe n’inyigo z’amashusho zigaragaza ko imiterere ya poroteyine nyinshi ikora ku rugero rwo hejuru rwa TMP na poroteyine ruri hejuru ya 200 mg/mL.

Uburyo bwiza burimo gukoresha sisitemu hafi ya TMP y'ingenzi, ariko ntirenze. Kuri iyi ngingo, umusaruro uba mwinshi ariko ibyago byo kwangirika bidasubirwaho biracyari bike. Ku bijyanye n'ubucucike buri hejuru cyane, ibyavumbuwe vuba aha bigaragaza ko kugabanya TMP no kongera urujya n'uruza rw'ibiryo icyarimwe (transverse flow filtration) kugira ngo bifashe kugabanya polarisation y'ibice no gushyira poroteyine mu buryo butunguranye. Urugero, ubushakashatsi ku bucucike bwa poroteyine ya Fc-fusion bwerekanye ko imiterere ya TMP iri hasi ifasha mu kugumana flux ihamye mu gihe bigabanya igihombo cy'umusaruro.

Kongera buhoro buhoro ingano ya poroteyine mu gihe cyo kuyungurura ni ingenzi cyane. Intambwe z’ingufu zishobora gutuma umuti ugera mu buryo bworoshye cyane, bikongera ibyago byo kwiteranya ndetse n’ubukana bw’ivumbikwa. Ahubwo, kongera ingano ya poroteyine buhoro buhoro bituma ibipimo by’imikorere nka TMP, umuvuduko w’amazi, na pH bihindurwa icyarimwe, bigafasha mu kubungabunga umutekano w’imikorere. Inyigo za enzymes ultrafiltration zemeza ko gukomeza kugabanya umuvuduko w’imikorere muri ibi byiciro bituma ubwiyongere bw’umuvuduko bugenzurwa, bigagabanya kugabanuka k’umuvuduko w’amazi mu gihe bikingira ubuziranenge bw’umusaruro.

6.2. Inshuro zo gusimbuza uturemangingo n'uburyo bwo kubungabunga

Inshuro zo gusimbuza uturemangingo mu kuyungurura imirambo bifitanye isano rikomeye n'ibimenyetso by'uko amazi yanduye n'uko amazi yagabanuka. Aho kwishingikiriza gusa ku kugabanuka k'amazi nk'ikimenyetso cy'iherezo ry'ubuzima, kugenzura uburyo amazi yanduye adashobora kugabanuka—igipimo cy'ingano kigaragaza ko amazi yanduye adashobora kugabanuka bitewe n'ibintu byakusanyijwe—byagaragaye ko byizewe cyane, cyane cyane mu biryo bya poroteyine cyangwa poroteyine-polysaccharide, aho amazi yanduye ashobora kubaho vuba kandi bikomeye.

Gukurikirana ibimenyetso by'inyongera by'isukura nabyo ni ingenzi cyane. Ibimenyetso bigaragara by'uko ubuso buhagaze, amazi atemba mu kirere adahuye, cyangwa kwiyongera guhoraho kwa TMP (nubwo hakorwa isuku) byose ni ibimenyetso by'uko isukura igeze kure mbere y'uko isukura. Ubuhanga nko gukurikirana igipimo cyahinduwe cy'isukura (MFI-UF) no kugihuza n'imikorere ya isukura bituma habaho gahunda yo gusimbuza aho guhindura ibintu, bityo bigabanye igihe cyo kudakora no kugenzura ikiguzi cyo kubungabunga.

Ubuziranenge bw'ibice by'umubiri bubangamirwa no kwiyongera kw'ibice by'umubiri, ndetse no kwangirika, cyane cyane mu mikorere ikora kuri pH ikabije cyangwa ifite umunyu mwinshi. Hagomba gushyirwaho igenzura rihoraho n'isukura rya shimi kugira ngo hagenzurwe ingese n'ibice by'ibice by'umubiri. Iyo hagaragaye ko ibice by'umubiri bivanze n'ingese, hagomba guhindurwa inshuro zo gusukura ibice by'umubiri n'igihe cyo kubisimbura kugira ngo habeho igihe kirekire cyo kumara ibice by'umubiri kandi imikorere yabyo irusheho kumera neza. Kubungabunga neza no kugena gahunda ni ingenzi kugira ngo bigabanye ingaruka z'ibi bibazo no kongera imikorere myiza.

6.3. Igenzura ry'imikorere n'ibipimo by'ubucucike bw'umurongo

Gupima neza kandi mu buryo nyabwo ubukana bwa poroteyine ni ingenzi mu kugenzura ibikorwa mu kuyungurura poroteyine, cyane cyane uko ubunini n'ubukana byiyongera. Sisitemu zo gupima ubukana zikora ku murongo zitanga igenzura rihoraho, zigatanga ibitekerezo byihuse kandi zigatuma habaho impinduka zihindagurika ku bipimo bya sisitemu.

Ikoranabuhanga rishya ryahinduye imiterere y'uburyo bwo gupima poroteyine mu buryo bwa viscosity:

Raman Spectroscopy hamwe na Kalman Filtering: Isesengura rya Raman ry’igihe nyacyo, rishyigikiwe n’ibikoresho bya Kalman byagutse, rituma habaho gukurikirana neza ingano ya poroteyine n’imiterere yayo. Ubu buryo bwongera ubushobozi bwo kumva no gukora neza, bugashyigikira imikorere y’imikorere kugira ngo ingano ya ultrafiltration irusheho kwiyongera no kwinjira mu mubiri.

Visometri ya Kinematic Capillary Viscometry yikora: Ikoresheje mudasobwa ireba, iyi koranabuhanga ipima ubukana bw'umuti mu buryo bwikora, igatsinda amakosa yo mu maboko kandi igatanga igenzura risubirwamo, rikorwa mu buryo bwinshi mu nzira zitandukanye. Yemejwe ku miterere ya poroteyine isanzwe n'iy'ubwoko butandukanye kandi igabanya ingaruka mu gihe cyo kuyungurura ingano.

Ibikoresho bya Microfluidic Rheology: Sisitemu za microfluidic zitanga imiterere irambuye kandi ihoraho ya rheological, ndetse no ku bisubizo bya poroteyine bitari Newtonian kandi bifite ubukana bwinshi. Ibi ni ingenzi cyane mu gukora imiti, gushyigikira ingamba za ikoranabuhanga ryo gusesengura imikorere (PAT) no guhuza n'ibisubizo.

Kugenzura ibikorwa hakoreshejwe ibi bikoresho bituma habaho ishyirwa mu bikorwa rya “feedback loops” kugira ngo hakosorwe TMP mu buryo bwihuse, igipimo cy’ibiryo, cyangwa umuvuduko w’amazi mu gihe cy’impinduka mu buryo bw’ubushyuhe. Urugero, niba uburyo bwo kumenya ibintu buri mu murongo bubonye izamuka ritunguranye rya “viscosity” (bitewe no kwiyongera kw’ibipimo cyangwa kwishyira hamwe), TMP ishobora kugabanuka mu buryo bwikora cyangwa umuvuduko w’amazi mu buryo bw’ubushyuhe kugira ngo igabanye gutangira kwa “concentration polarization” mu gihe cyo kuyungurura cyane. Ubu buryo ntibwongera igihe cyo kubaho gusa ahubwo bunashyigikira ubwiza bw’ibicuruzwa mu buryo buhoraho binyuze mu gucunga ibintu bigira ingaruka ku buryo bufatika bwa “viscosity” ya poroteyine.

Guhitamo ikoranabuhanga rikwiye ryo kugenzura ubucucike biterwa n'ibikenewe byihariye mu ikoreshwa rya ultrafiltration, harimo ingano y'ubucucike bwitezwe, uburyo poroteyine zitunganywamo, ibikenewe mu guhuza, n'igiciro. Iri terambere mu igenzura ry'igihe nyacyo no kugenzura inzira zihinduka ryazamuye cyane ubushobozi bwo kunoza uburyo poroteyine zitunganywamo cyane, bishimangira ko imikorere yazo ihamye kandi igatanga umusaruro mwinshi.

Gukemura ibibazo n'ibibazo bisanzwe mu bijyanye no kuyungurura poroteyine

7.1. Ibimenyetso, Impamvu, n'Imiti

Ubwiyongere bw'umuvuduko wa Transmembrane

Ukwiyongera k'umuvuduko wa transmembrane (TMP) mu gihe cyo kuyungurura imirambo bigaragaza ko imbaraga zikomeza kwiyongera mu ruhu. Ingaruka z'umuvuduko wa transmembrane ku kuyungurura imirambo ni izitaziguye: urwego rusanzwe rw'umuvuduko wa transmembrane akenshi rushingira ku mikorere, ariko kwiyongera guhoraho ni ngombwa gusuzuma. Impamvu ebyiri zikunze kugaragara:

• Ubushyuhe bwinshi bw'umuti wa poroteyine:Uko ubushyuhe bw'ibisubizo bya poroteyine bwiyongera—akenshi iyo poroteyine nyinshi ziyungururwa cyane—umuvuduko ukenewe kugira ngo amazi atemba wiyongere. Ibi bigaragara mu ntambwe za nyuma zo gucukura no gucukura aho ibisubizo biba bikomeye cyane.
• Guhumanya uturemangingo:Udukoko nk'udukoko twa poroteyine cyangwa uruvange rwa polysaccharide-proteine ​​dushobora gufata cyangwa kuziba imyenge y'ururabo, bigatuma TMP yiyongera cyane.

Imiti:

• Kugabanya TMP no kongera ubwinshi bw'ibiryo: Kugabanya TMP mu gihe wongera umuvuduko w'ibiryo bigabanya polarisation y'ibipimo n'imiterere y'urutonde rwa gel, bigatera flux idahindagurika.
• Gusukura uturemangingo buri gihe: Shyiraho uburyo bwiza bwo gusukura uturemangingo kugira ngo ukureho ibintu bibi byakusanyije. Genzura uburyo bikoreshwa ukoresheje uburyo bwo gupima ubukana bwa poroteyine nyuma yo gusukura.
• Simbuza uturemangingo tw'umubiri dusaza: Inshuro nyinshi zo gusimbuza uturemangingo zishobora kuba ngombwa niba isuku idahagije cyangwa igihe cyo kumara uturemangingo kigeze.

Kugabanuka k'igipimo cy'ihindagurika ry'ihindagurika: Igipimo cyo gusuzuma

Kugabanuka guhoraho k'umuvuduko mu gihe cy'icyiciro cya ultrafiltration bigaragaza impungenge z'umusaruro. Kurikiza ubu buryo bwo gusuzuma:

1. Kugenzura TMP n'ubukana:Niba byombi byiyongereye, reba niba hari umwanda cyangwa urwego rwa gel rurimo.
2. Suzuma imiterere y'ibiryo n'ubuziranenge bwa pH:Guhinduranya ibintu hano bishobora guhindura ubukana bw'ibisubizo bya poroteyine no guteza umwanda.
3. Suzuma imikorere y'ururenda:Kugabanuka k'urujya n'uruza rw'amazi nubwo hashobora gukurwaho ibimenyetso by'uko uturemangingo dushobora kwangirika cyangwa kwangirika bidasubirwaho.

Ibisubizo:

• Ongera ubushyuhe, pH, n'imbaraga za iyoni mu biryo kugira ngo ugabanye umwanda no guhindagurika kw'ibipimo mu gihe cyo kuyungurura cyane.
• Koresha modules zahinduwe ku buso cyangwa zizunguruka kugira ngo uhagarike imiterere ya gel kandi ugarure flux.
• Kora buri gihe gupima ubukana bwa poroteyine kugira ngo umenye impinduka zigira ingaruka ku rujya n'uruza rw'amazi.

Guhindura ibara ry'uruhu vuba cyangwa imiterere ya Gel Layer

Gushingwa vuba k'urwego rwa gel bituruka ku guhindagurika cyane kw'ibipimo ku buso bwa membrane. Igitutu cya transmembrane cya transverse flow filtration (TFF) gikunze kwibasirwa cyane cyane iyo hari imiterere myinshi cyangwa poroteyine nyinshi.

Ingamba zo kugabanya ingaruka:

• Shyira ku buso bw'urukiramende rudakoresha amazi kandi rufite umuriro ukabije (urugero, uturemangingo twa Polyvinylidene fluoride [PVDF]) kugira ngo ugabanye gufatana no gufatana kwa poroteyine.
• Kugaburira mbere ukoresheje uburyo bwo gufungana cyangwa gukoresha amashanyarazi kugira ngo ukureho ibintu byinshi byangiza mbere yo kuyungurura cyane.
• Shyiramo ibikoresho bya mekanike nka modules zizunguruka mu buryo bwo kuyungurura amazi mu buryo bwo kugabanya ubugari bw'urutonde rw'ibice bya keke no gutinza ikorwa ry'urutonde rwa gel.

7.2. Guhindura uburyo bwo gutanga ibiribwa bihinduka

Uburyo bwo kuyungurura poroteyine bugomba guhinduka bitewe n’imiterere cyangwa imiterere ya poroteyine y’ibiryo. Ibintu bigira ingaruka ku buryo poroteyine ikora—nk’imiterere y’ibinyabutabire, ubwinshi bwa poroteyine, n’uburyo bwo guteranya—bishobora guhindura imikorere y’urusobe rw’ibintu.

Ingamba zo Gusubiza

• Gukurikirana ubukana n'imiterere y'ikintu mu gihe nyacyo:Shyiraho ibikoresho byo gusesengura biri kuri interineti (Raman spectroscopy + Kalman filtering) kugira ngo bigaragare vuba impinduka mu miyoboro y'amazi, birushe uburyo bwa kera bwa UV cyangwa IR.
• Igenzura ry’imikorere ijyanye n’igihe:Hindura igenamiterere rya parametero (igipimo cy'amazi anyuramo, TMP, guhitamo membrane) mu rwego rwo gusubiza impinduka zabonetse. Urugero, kwiyongera kwa poroteyine mu buryo bw'umusemburo bishobora gusaba TMP nkeya no gukata cyane.
• Guhitamo uturemangingo:Koresha uturemangingo dufite ingano y'imyenge n'ubuso bwa shimi byakozwe neza kugira ngo bifashe mu miterere y'ibiryo, kuringaniza uburyo poroteyine zibikwa n'uburyo zigenda.
• Uburyo bwo kugaburira mbere yo kugaburira:Niba impinduka zitunguranye mu mirire y'ibiryo zituma habaho umwanda, shyiramo intambwe zo gufungana cyangwa kuyungurura zizamuka mu gihe cyo kuyungurura cyane.

Ingero:

• Mu gutunganya ibintu bishya, guhinduranya ibintu mu buryo bwa buffer cyangwa impinduka mu mikorere y’uturemangingo tw’umubiri bigomba gutuma habaho impinduka za TMP n’imigendekere y’amaraso binyuze muri sisitemu yo kugenzura.
• Ku bijyanye na chromatography-linked ultrafiltration, adaptive mixing-integer optimization algorithms ishobora kugabanya ihindagurika no kugabanya ikiguzi cy'imikorere mu gihe ikomeza imikorere ya ultrafiltration membrane.

Gukurikirana buri gihe uburyo bwo gupima ubukana bwa poroteyine no guhindura imiterere y’imikorere y’ibice bifasha kunoza uburyo bwo kuyungurura ingano y’ibice binini, gukomeza kohereza umusaruro, no kugabanya kwangirika kw’ibice binini n’ubwiyongere bw’ibice binini.

Ibibazo Bikunze Kubazwa

8.1. Ni uruhe rugero rusanzwe rw'umuvuduko wa transmembrane mu gusuzuma poroteyine mu buryo bwa ultrafiltration?

Igipimo gisanzwe cy’umuvuduko wa transmembrane (TMP) muri sisitemu yo kuyungurura poroteyine giterwa n’ubwoko bw’urukiramende, imiterere ya module, n’imiterere y’ibiryo. Ku buryo bwinshi bwo kuyungurura poroteyine, TMP ikunze kuba hagati ya bar 1 na 3 (15–45 psi). Agaciro ka TMP kari hejuru ya 0.2 MPa (hafi 29 psi) gashobora kwangiza urukiramende, kwangiza vuba, no kugabanya igihe cy’urukiramende. Mu bikorwa byo mu rwego rw’ubuvuzi no gutunganya ibinyabuzima, TMP isabwa muri rusange ntigomba kurenza bar 0.8 (~ 12 psi) kugira ngo hirindwe ko urukiramende rwacika. Ku bikorwa nko kuyungurura amashanyarazi, kuguma muri uru rwego rwa TMP birinda umusaruro no kuba poroteyine ikora neza.

8.2. Ni gute ubukana bw'ibisubizo bya poroteyine bigira ingaruka ku mikorere ya ultrafiltration?

Ubucucike bwa poroteyine bugira ingaruka zitaziguye ku mikorere y’ubwinshi bw’ibice bya ultrafiltration. Ubucucike bwinshi byongera ubudahangarwa bw’amazi kandi bukazamura TMP, bigatuma flux igabanuka kandi ikava mu ruhu rw’umuyoboro w’amaraso ikava mu ruhu ikagira ingaruka mbi ku ruhu. Iyi ngaruka igaragara iyo utwo dukoko tw’umubiri twitwa monoclonal antibody cyangwa poroteyine za Fc-fusion ziri mu bwinshi, aho ubucucike bwiyongera bitewe n’imikoranire ya poroteyine na poroteyine n’ingaruka za charge. Gucunga no kunoza ubucucike hakoreshejwe ibikoresho bifasha mu gukurura cyangwa enzymes binoza flux, bigabanya ubucucike, kandi bigatuma flux iboneka cyane mu gihe cy’ubwinshi bw’ibice bya ultrafiltration. Gukurikirana gupima ubucucike bwa poroteyine ni ingenzi kugira ngo hakomeze gutunganywa neza.

8.3. Guhuza imbaraga mu buryo bw'ingufu (concentration polarization) ni iki kandi kuki ari ingenzi muri TFF?

Guhuza imiterere y’ibice mu gucukura ikirere ni ukwirundanya kwa poroteyine ku buso bw’urukiramende, bigatera intera iri hagati y’umusaruro mwinshi n’aho urukiramende ruhurira. Mu gucukura imiterere y’amazi mu buryo bunyuranyije n’umurongo, ibi bitera kwiyongera kwa viscosity yo mu gace ndetse no kugabanuka kw’amazi mu buryo bushobora gusubira inyuma. Iyo bitagenzuwe, bishobora guteza ihindagurika ry’imiterere y’urukiramende no kugabanya imikorere myiza y’urukiramende. Gukemura ikibazo cyo guhuza imiterere y’ibice mu gucukura ikirere bikubiyemo kunoza umuvuduko w’amazi mu gucukura ikirere, TMP, no guhitamo imiterere y’urukiramende kugira ngo hakomeze kuba urwego rworoshye rwo guhuza imiterere y’ibice. Kugenzura neza bikomeza ko umusaruro unyura hejuru kandi ibyago byo kuzana ihindagurika ry’ikirere bikaba bike.

8.4. Ni gute nafata icyemezo cyo gusimbuza membrane yanjye ya ultrafiltration?

Simbuza membrane ya ultrafiltration iyo ubonye igabanuka rigaragara ry'umuvuduko w'amazi (flux), ukwiyongera guhoraho kwa TMP kudashobora gukemura isuku isanzwe, cyangwa umwanda ugaragara usigara nyuma yo gusukurwa. Ibindi bimenyetso birimo gutakaza ubushobozi bwo guhitamo (kunanirwa kwanga poroteyine zigenewe nk'uko byari byitezwe) no kudashobora kugera ku bipimo by'imikorere. Gukurikirana inshuro zo gusimbuza membrane hakoreshejwe flux isanzwe no gupima ubushobozi bwo guhitamo ni ishingiro ryo kongera igihe cyo kubaho kwa membrane mu mikorere ya protein ultrafiltration.

8.5. Ni ibihe bipimo by'imikorere nshobora guhindura kugira ngo gabanyirize poroteyine muri TFF?

Ibipimo by'ingenzi by'imikorere kugira ngo bigabanye kwanduza poroteyine mu kuyungurura amazi hakoreshejwe ikoranabuhanga birimo:

• Gukomeza umuvuduko uhagije wo kunyura mu biryo kugira ngo ugabanye ukwiyongera kwa poroteyine mu gace utuyemo no gucunga ubwiyongere bw'ibinure.
• Koresha mu rugero rwa TMP rwasabwe, ubusanzwe hagati ya psi 3–5 (0.2–0.35 bar), kugira ngo wirinde ko ibicuruzwa bisohoka cyane kandi bikangiza uturemangingo.
• Koresha amabwiriza yo gusukura membrane buri gihe kugira ngo ugabanye umwanda udakira.
• Genzura kandi, nibiba ngombwa, banza ushyire umuti w’ibiryo mu rwego rwo kugenzura ubushyuhe bw’ibiryo (urugero, ukoresheje imiti igabanya ubushyuhe nka pectinase).
• Hitamo ibikoresho bya membrane n'ingano y'imyenge (MWCO) bikwiranye n'ingano ya poroteyine n'intego z'imikorere.

Gushyira hamwe uburyo bwo kuyungurura hydrocyclone cyangwa uburyo bwo kuyungurura enzyme bishobora kunoza imikorere ya sisitemu, cyane cyane ku biryo bifite ubukana bwinshi. Kugenzura neza imiterere y'ibiryo no guhindura imiterere yabyo mu buryo buhamye kugira ngo ugabanye kwangirika kw'ibiryo no kunoza icyiciro cyo kuyungurura ultrafiltration.