Iindaba - Ukubek'esweni amaNqanaba e-pH kwiNkqubo ye-Bio Pharmaceutical Fermentation

I-electrode ye-pH idlala indima ebalulekileyo kwinkqubo yokuvutshelwa, ngokuyinhloko ikhonza ukubeka iliso kunye nokulawula ubumuncu kunye ne-alkalinity yomhluzi wokubilisa. Ngokuqhubekayo ukulinganisa ixabiso le-pH, i-electrode yenza ulawulo oluchanekileyo kwindawo yokubilisa. I-electrode ye-pH eqhelekileyo iquka i-electrode yokuva kunye ne-electrode yereferensi, esebenza kumgaqo we-Nernst equation, elawula ukuguqulwa kwamandla ekhemikhali abe ziimpawu zombane. Ikhono le-electrode lihambelana ngokuthe ngqo nomsebenzi we-hydrogen ions kwisisombululo. Ixabiso le-pH limiselwa ngokuthelekisa umahluko olinganisiweyo wamandla ombane kunye neso sisombululo siqhelekileyo sesithinteli, sivumela ukulinganisa okuchanekileyo nokuthembekileyo. Le ndlela yokulinganisa iqinisekisa ukulawulwa kwe-pH ezinzileyo kuyo yonke inkqubo yokuvutshelwa, ngaloo ndlela ixhasa umsebenzi ophezulu we-microbial okanye weselula kunye nokuqinisekisa umgangatho wemveliso.

Ukusetyenziswa ngokufanelekileyo kwee-electrodes ze-pH kufuna amanyathelo amaninzi okulungiselela, kubandakanywa nokusebenza kwe-electrode-ngokuqhelekileyo kufezekiswa ngokuntywiliselwa kwe-electrode emanzini adibeneyo okanye isisombululo se-pH 4 se-buffer-ukuqinisekisa ukuphendula ngokufanelekileyo kunye nokuchaneka kokulinganisa. Ukuhlangabezana neemfuno ezingqongqo zeshishini le-biopharmaceutical fermentation, ii-electrode ze-pH kufuneka zibonise amaxesha okuphendula akhawulezayo, ukuchaneka okuphezulu, kunye nokomelela phantsi kweemeko ezingqongqo zokuvala inzala ezifana ne-high-temperature sterilization (SIP). Ezi mpawu zivumela ukusebenza okuthembekileyo kwiindawo ezingcolileyo. Umzekelo, kwimveliso ye-glutamic acid, ukujonga okuchanekileyo kwe-pH kubalulekile ekulawuleni iiparamitha eziphambili ezifana nobushushu, i-oksijini enyibilikileyo, isantya sokuphazamiseka, kunye ne-pH ngokwayo. Ukulawulwa ngokuchanekileyo kwezi zintlukwano kuchaphazela ngokuthe ngqo isivuno kunye nomgangatho wemveliso yokugqibela. Ezinye ii-electrode ze-pH eziphucukileyo, ezineembrane zeglasi ezikwaziyo ukumelana nobushushu obuphezulu kunye neenkqubo zesalathiso zejeli zepolymer ezicinezelekileyo, zibonisa uzinzo olukhethekileyo phantsi kobushushu obugqithisileyo kunye neemeko zoxinzelelo, nto leyo ezenza ukuba zilungele ukusetyenziswa kwe-SIP kwiinkqubo zebhayoloji kunye nokutya kokutya. Ngaphaya koko, izakhono zabo eziqinileyo zokuchasana nokungcolisa zivumela ukusebenza okungaguquguqukiyo kuzo zonke iibroths ezahlukeneyo zokubilisa. I-Shanghai Boqu Instrument Co., Ltd. inikezela ngeendlela ezahlukeneyo zokudityaniswa kwe-electrode, ukuphucula lula komsebenzisi kunye nokuguquguquka kokuhlanganiswa kwenkqubo.

Kutheni i-pH esweni iyimfuneko ngexesha lenkqubo yokuvundiswa kwe-biopharmaceuticals?

Kwi-biopharmaceutical fermentation, ukujonga ixesha lokwenyani kunye nokulawulwa kwe-pH kubalulekile kwimveliso eyimpumelelo kunye nokwandisa isivuno kunye nomgangatho weemveliso ezijoliswe kuyo ezifana ne-antibiotics, izitofu zokugonya, i-monoclonal antibodies, kunye ne-enzymes. Ngokwenyani, ulawulo lwe-pH ludala imeko-bume efanelekileyo yendalo ye-microbial okanye iiseli ezanyisayo-ezisebenza "njengemizi-mveliso ephilayo"-ukukhula kunye nokudibanisa iikhompawundi zonyango, ngendlela efanayo nendlela amafama ahlengahlengisa ngayo i-pH yomhlaba ngokweemfuno zezityalo.

1. Gcina umsebenzi weselula uphezulu
Ukuvubela kuxhomekeke kwiiseli eziphilayo (umzekelo, iiseli ze-CHO) ukuvelisa i-biomolecules enzima. Imetabolism yeselula inovelwano kakhulu kwi-pH yokusingqongileyo. I-Enzymes, ezenza zonke ii-intracellular biochemical reactions, zine-pH optima emxinwa; ukuphambuka kolu luhlu kunokunciphisa kakhulu umsebenzi we-enzymatic okanye kubangele i-denaturation, ukuphazamisa umsebenzi we-metabolic. Ukongeza, ukunyuswa kwezondlo nge-membrane yeseli-efana ne-glucose, i-amino acids, kunye neetyuwa ze-inorganic-ixhomekeke kwi-pH. Amanqanaba e-pH asezantsi anokuthintela ukufunxwa kwezondlo, okukhokelela ekukhuleni okungafanelekanga okanye ukungalingani kwe-metabolic. Ngapha koko, amaxabiso agqithisileyo e-pH anokuthi abeke esichengeni ingqibelelo ye-membrane, okukhokelela ekuvuzeni kwe-cytoplasmic okanye i-cell lysis.

2. Ukunciphisa ukubunjwa kwemveliso kunye nenkunkuma ye-substrate
Ngexesha lokuvutshelwa, i-metabolism yeselula ivelisa i-acidic okanye i-metabolites esisiseko. Ngokomzekelo, ii-microorganisms ezininzi zivelisa i-organic acids (umzekelo, i-lactic acid, i-acetic acid) ngexesha le-glucose catabolism, ebangela ukuhla kwe-pH. Ukuba ayilungiswanga, i-pH ephantsi inqanda ukukhula kweeseli kwaye inokutshintsha i-metabolic flux iye kwiindlela ezingavelisi mveliso, inyusa ukuqokelelana kwemveliso. Ezi mveliso zisebenzisa ikhabhoni kunye nezibonelelo zamandla ezinokuthi zixhase ukuhlanganiswa kwemveliso ekujoliswe kuyo, ngaloo ndlela kuncitshiswe isivuno sonke. Ulawulo lwe-pH olusebenzayo lunceda ukugcina iindlela ezinqwenelekayo zemetabolism kunye nokuphucula ukusebenza kakuhle kwenkqubo.

3. Qinisekisa ukuzinza kwemveliso kwaye uthintele ukuthotywa
Iimveliso ezininzi ze-biopharmaceutical, ngakumbi iiprotheyini ezinjenge-monoclonal antibodies kunye ne-peptide hormones, zichaphazeleka kwiinguqu ze-pH-induced structure. Ngaphandle koluhlu lwe-pH oluzinzileyo, ezi molekyuli zinokungena kwi-denaturation, ukuhlanganiswa, okanye ukungasebenzi, okunokuthi zenze imvula eyingozi. Ukongeza, iimveliso ezithile ziqhelene nekhemikhali ye-hydrolysis okanye ukuthotywa kwe-enzymatic phantsi kweemeko ze-acidic okanye zealkaline. Ukugcina i-pH efanelekileyo kunciphisa ukuthotywa kwemveliso ngexesha lokuvelisa, ukugcina amandla kunye nokhuseleko.

4. Ukwandisa ukusebenza kakuhle kwenkqubo kunye nokuqinisekisa ukuhambelana kwebhetshi ukuya kwibhetshi
Ukusuka kwimboniselo, ulawulo lwe-pH luchaphazela ngokuthe ngqo imveliso kunye nokuhlala kakuhle kwezoqoqosho. Uphando olubanzi luqhutyelwa ukuchonga iindawo ezifanelekileyo ze-pH zezigaba ezahlukeneyo zokuvutshelwa-ezifana nokukhula kweeseli ngokubhekiselele kwimveliso yemveliso-enokuthi ihluke kakhulu. Ulawulo lwe-pH eDynamic luvumela ukulungelelaniswa kwenqanaba elithile, ukwandisa ukuqokelelwa kwe-biomass kunye namanqaku emveliso. Ngapha koko, ii-arhente ezilawulayo ezifana ne-FDA kunye ne-EMA zifuna ukuthotyelwa ngokungqongqo kwiZiqhelo zokuVelisa eziLungileyo (GMP), apho iiparamitha zenkqubo ehambelanayo zinyanzelekile. I-pH ibonwa njengeParameter yeNkqubo eBalulekileyo (CPP), kwaye ukubeka iliso ngokuqhubekayo kuqinisekisa ukuveliswa kwakhona kwiibhetshi, ukuqinisekisa ukhuseleko, ukusebenza kakuhle, kunye nomgangatho weemveliso zamachiza.

5. Khonza njengesalathisi sempilo yokuvutshelwa
Umkhwa wokutshintsha kwe-pH ubonelela ngeembono ezixabisekileyo kwimeko ye-physiological yenkcubeko. Ukutshintsha ngokukhawuleza okanye okungalindelekanga kwi-pH kunokubonisa ungcoliseko, ukungasebenzi kakuhle kwenzwa, ukuncipha kwezondlo, okanye ukungahambi kakuhle kwe-metabolic. Ukufunyaniswa kwangoko okusekwe kwiindlela ze-pH kuvumela ukungenelela kwabasebenzi ngexesha, ukuququzelela ukulungisa iingxaki kunye nokuthintela ukusilela kwebhetshi eneendleko.

Ingaba izinzwa ze-pH kufuneka zikhethwe njani kwinkqubo yokuvubela kwi-biopharmaceuticals?

Ukukhetha i-sensor ye-pH efanelekileyo ye-biopharmaceutical fermentation sisigqibo esibalulekileyo sobunjineli esichaphazela ukuthembeka kwenkqubo, ukuthembeka kwedatha, umgangatho wemveliso, kunye nokuthotyelwa kokulawula. Ukhetho kufuneka lusondele ngokucwangcisiweyo, lungaqwalaselwanga kuphela ukusebenza kwenzwa kodwa kunye nokuhambelana nokuhamba komsebenzi wonke we-bioprocessing.

1. Ubushushu obuphezulu kunye nokumelana noxinzelelo
Iinkqubo ze-Biopharmaceutical zivame ukusebenzisa i-in-situ sterilization (SIP), ngokuqhelekileyo kwi-121 ° C kunye ne-1-2 bar yoxinzelelo lwe-20-60 imizuzu. Ke ngoko, nayiphi na inzwa ye-pH kufuneka imelane nokuvezwa ngokuphindaphindiweyo kwiimeko ezinjalo ngaphandle kokusilela. Ngokufanelekileyo, inzwa kufuneka ilinganiswe ubuncinane kwi-130 ° C kunye ne-3-4 bar ukubonelela ngomda wokhuseleko. Ukutywinwa okuqinileyo kubalulekile ukukhusela ukungena komswakama, ukuvuza kwe-electrolyte, okanye ukonakala koomatshini ngexesha lokuhamba ngebhayisikile eshushu.

2. Uhlobo lwenzwa kunye nenkqubo yereferensi
Le yingqwalasela engundoqo yobugcisa echaphazela uzinzo lwexesha elide, iimfuno zokugcinwa, kunye nokuchasana nokungalunganga.
Ukucwangciswa kwe-Electrode: I-electrodes edibeneyo, ukudibanisa zombini izinto zokulinganisa kunye nereferensi kumzimba omnye, zamkelwa ngokubanzi ngenxa yokulula ukufakwa kunye nokuphatha.
Inkqubo yoReferensi:
• Ireferensi ezaliswe lulwelo (umzekelo, isisombululo se-KCl): Inikezela ngempendulo ekhawulezayo kunye nokuchaneka okuphezulu kodwa ifuna ukuzaliswa kwakhona ngamaxesha athile. Ngexesha le-SIP, ukulahleka kwe-electrolyte kunokwenzeka, kwaye i-porous junctions (umz., i-ceramic frits) ijwayele ukuvaleka ngeeprotheni okanye i-particulates, ekhokelela ekufundweni kunye nokungathembeki.
• Ijeli yepolymer okanye ireferensi yesimo esiqinileyo: Ithandwa kakhulu kwii-bioreactors zanamhlanje. Ezi nkqubo ziphelise imfuno yokuzaliswa kwakhona kwe-electrolyte, ukunciphisa ukugcinwa, kunye nokudibanisa ulwelo olubanzi (umzekelo, amakhonkco e-PTFE) axhathisa ukungcola. Banikezela ngozinzo oluphezulu kunye nobomi benkonzo obude kwimidiya entsonkothileyo, i-viscous fermentation media.

3. Uluhlu lomlinganiselo kunye nokuchaneka
Inzwa kufuneka igubungele uluhlu olubanzi lokusebenza, ngokuqhelekileyo pH 2-12, ukulungiselela izigaba zenkqubo ezahlukeneyo. Ukubona ubuntununtunu beenkqubo zebhayoloji, ukuchaneka komlinganiselo kufuneka kube ngaphakathi kwe-±0.01 ukuya kwi-±0.02 yeeyunithi ze-pH, ezixhaswa yimveliso yesignali yesisombululo esiphezulu.

4. Ixesha lokuphendula
Ixesha lokuphendula lichazwa ngokuqhelekileyo njenge-t90-ixesha elifunekayo ukufikelela kwi-90% yokufundwa kokugqibela emva kokutshintshwa kwesinyathelo kwi-pH. Ngelixa ii-electrode zodidi lwejeli zinokubonisa impendulo ecothayo kunezo zizaliswe lulwelo, ngokubanzi ziyahlangabezana neemfuno eziguquguqukayo zolawulo lwelophu yokulawula ukubila, esebenza ngokwemilinganiselo yeyure kunesesekondi.

5. Biocompatibility
Zonke izinto ezinxibelelana nenkcubeko yenkcubeko kufuneka zingabi yityhefu, zingabi yityhefu, kwaye zingasebenzi ukuthintela iziphumo ezibi ekusebenzeni kweeseli okanye umgangatho wemveliso. Iintlobo zeglasi ezikhethekileyo ezenzelwe izicelo ze-bioprocessing ziyacetyiswa ukuqinisekisa ukuxhathisa imichiza kunye ne-biocompatibility.

6. Imveliso yesignali kunye nojongano
• Imveliso ye-analog (mV/pH): Indlela yemveli isebenzisa ukuhanjiswa kwe-analog kwinkqubo yokulawula. Ixabiso elisebenzayo kodwa lisengozini yokuphazamiseka kwe-electromagnetic kunye nokuthomalalisa umqondiso kwimigama emide.
• Imveliso yedijithali (umzekelo, i-MEMS-based okanye i-smart sensors): Idibanisa i-microelectronics ekwibhodi yokuhambisa imiqondiso yedijithali (umz., nge-RS485). Ibonelela ngokhuseleko lwengxolo olugqwesileyo, ixhasa unxibelelwano lomgama omde, kwaye yenza ukuba ukugcinwa kwembali yolungelelwaniso, iinombolo zeserial, kunye neelog zokusebenzisa. Ithobelana nemigangatho yolawulo efana ne-FDA 21 CFR iSigaba se-11 malunga neerekhodi ze-elektroniki kunye nemisayino, iyenza ithandeke ngakumbi kwiindawo ze-GMP.

7. I-interface yokufakela kunye nezindlu ezikhuselayo
Isivamvo kufuneka sihambelane nezibuko elichongiweyo kwi-bioreactor (umzekelo, i-tri-clamp, i-sanitary fitting). Imikhono ekhuselayo okanye oonogada bayacetyiswa ukuba bathintele umonakalo owenziwe ngoomatshini ngexesha lokuphatha okanye ukusebenza kunye nokwenza lula ukutshintshwa ngaphandle kokubeka esichengeni ubunyumba.