Madal Horumarinta Antibody-ga Bispecific

Alpha Lifetech waxay macaamiisha siin kartaa helitaan tayo leh oo antibody ah iyo badeecooyin iyo adeegyo borotiin dib-u-habeyn ah. Waxaan diyaarin karnaa antibody-yo leh waxtar sare, gaar ahaansho xooggan, iyo xasillooni wanaagsan. Alpha Lifetech waxay leedahay qalab iyo qalab kala duwan oo nadiifinta antibody-yo ah, kuwaas oo bixin kara adeegyo nadiifinta antibody-yo ah oo laga helo ilo kala duwan sida antibody-yada bakaylaha, idaha, digaagga, iyo jiirka monoclonal-ka, iyo sidoo kale adeegyada nadiifinta Protein A/G affinity iyo adeegyada kala-soocidda antibody-yo iyo nadiifinta. Iyada oo ku saleysan dhismaha nidaamka dhamaystiran ee madal-ka ee madal-ka helitaanka antibody-yo, madal borotiin, iwm., waxaan daboolnaa adeegyada kor iyo hoos ee wax soo saarka antibody-yo, waxaanan bixin karnaa adeegyo farsamo oo ka imanaya diyaarinta antibody-yo, nadiifinta antibody-yo laba-geesoodka ah iyo kala-soocidda antibody-yo iyo nadiifinta antibody-yo, taxanaha antibody-yo, xaqiijinta antibody-yo, iwm., kuwaas oo loo isticmaali karo daaweynta antibody-yo laba-geesoodka ah.

Unugyada difaaca jirka ee monoclonal waxay tilmaamayaan immunoglobulins-ka ay soo saarto hal unug oo B ah oo leh gaar ahaansho sare oo loogu talagalay antigens-ka ama epitopes-ka. Soo saarista unugyada difaaca jirka ee monoclonal waxaa markii hore lagu diyaariyey iyadoo la adeegsanayo tiknoolajiyadda hybridoma si loo soo saaro unugyada difaaca jirka ee jiirka. Gaar ahaan, iyadoo la isku darayo unugyada beeryarada ee jiirka la tallaalay oo leh unugyada myeloma ee aadanaha ama jiirka, unugyada hybridoma ayaa la sameeyaa, kuwaas oo soo saara unugyada difaaca jirka ee gaarka ah. Unugyada difaaca jirka ee monoclonal-ka ee jiirka ee la soo saaray waxaa inta badan loo isticmaalaa cilmi-baarista xayawaanka iyo ogaanshaha cudurka ka dib adeegyada nadiifinta. Si kastaba ha ahaatee, jirka bini'aadamka wuxuu soo saari doonaa jawaab difaac oo ku saabsan borotiinada jiirka ee shisheeye, sidaa darteed waxaa jira xaddidaadyo muhiim ah oo ku saabsan isticmaalka kiliinikada ee wax soo saarka unugyada difaaca jirka ee monoclonal-ka jiirka. Dabadeed, dadku waxay isticmaaleen "humanization" si ay u diyaariyaan unugyada difaaca jirka ee monoclonal ee jiirka, waxayna isticmaaleen tignoolajiyada injineernimada hidde-sidaha si ay wax uga beddelaan unugyada difaaca jirka ee jiirka si ay u yeeshaan gobol joogto ah oo immunoglobulin bini'aadamka ah si loo yareeyo difaacooda. Noocan antibody-ga waxaa sidoo kale loo yaqaan antibody monoclonal ah oo bini'aadam ah. Intaa waxaa dheer, isticmaalka unugyada aadanaha si loo soo saaro unugyada difaaca jirka ee monoclonal waxaa loo yaqaan diyaarinta antibody-ga monoclonal ee aadanaha oo dhan. Unugyada difaaca jirka ee monoclonal waxaa si weyn loogu isticmaalaa codsiyada biomedical iyo kiliinikada. Cilmi-baarista caafimaadka, unugyada difaaca jirka ee monoclonal waxaa loo isticmaali karaa sidii immunomodulators, unugyada difaaca jirka ee monoclonal-ka ee daaweynta waxaa loo isticmaali karaa kiimoterabiga kansarka iyo daaweynta caabuqa fayraska, iyo horumarinta unugyada difaaca jirka ee monoclonal-ka ayaa inta badan loo isticmaalaa daaweynta kansarka.

Hordhac ku saabsan Antibody-ga Bispecific

Sannadkii 1960, fikradda unugyada difaaca jirka ee bispecific ayaa la soo jeediyay. Unugyada difaaca jirka ee bispecific, oo sidoo kale loo yaqaan unugyada difaaca jirka ee monoclonal bispecific, waa unugyada difaaca jirka ee si macmal ah loo sameeyay iyadoo la adeegsanayo tignoolajiyada injineernimada hidde-sidaha. Maadaama ay tahay unug difaaca jirka ee macmalka ah oo la farsameeyay, unugyada difaaca jirka ee bispecific badanaa waxay ka tirsan yihiin fasalka hoose ee IgG waxayna ka kooban yihiin jajab isku xidhka antigen oo bartilmaameedsanaya qaybta hoose ee CD3. Unugyada difaaca jirka ee bispecific waxay leeyihiin laba goobood oo gaar ah oo isku xidha antigen kuwaas oo isku mar xiri kara oo aqoonsan kara laba antigen oo kala duwan, ama laba epitopes oo kala duwan oo antigen ah. Marka la barbardhigo unugyada difaaca jirka ee monoclonal, unugyada difaaca jirka ee bispecific waxay leeyihiin goob isku xidhka antigen oo gaar ah, sidaas darteedna leh gaar ahaansho xooggan iyo awood bartilmaameed, taas oo si sax ah u beegsan karta unugyada kansarka oo yareyn karta sunta bartilmaameedka. Unugyada difaaca jirka ee bispecific waxay isku mar samayn karaan hawlo badan oo bayooloji ah, sida qorista unugyada difaaca jirka, xannibaadda waddooyinka calaamadaynta, iyo si toos ah u dila unugyada burooyinka. Unugyada difaaca jirka ee bispecific hore waxaa inta badan lagu diyaariyey isku xidhka kiimikada ama isku-darka unugyada, laakiin habkani wuxuu si tartiib tartiib ah u socon karaa sababtoo ah isku-darka aan kala sooca lahayn iyo dhibka lagu qabo kala-goynta isku-darka bartilmaameedka. Iyadoo la horumarinayo tignoolajiyada injineernimada hidde-sidaha, goobo badan oo tignoolajiyadeed oo cusub ayaa la sameeyay, sida guntimaha godadka (KIH), CrossMab, DVD Ig, iwm. Goobahani waxay si wax ku ool ah u xalliyaan dhibaatooyinka sida isku dheelitir la'aanta silsiladda culus iyo silsiladda fudud, waxayna hagaajiyaan isku-dhafka iyo wax-soo-saarka unugyada difaaca jirka ee laba-geesoodka ah.

Jaantuska 1 Dulmar guud oo fudud oo ku saabsan hababka ficilka ee la soo jeediyay ee unugyada difaaca jirka ee laba-geesoodka ah (bsAbs) ee tijaabooyinka kiliinikada ee kansarka.(Isha sawirka: Dib u eegis ku saabsan unugyada difaaca jirka ee laba-geesoodka ah iyo qaab-dhismeedka unugyada difaaca jirka ee kansarka iyo caqabadaha caafimaad - ScienceDirect)

Tiknoolajiyada Soo-saarka Antibodies-ka Bispecific

Hababka ugu muhiimsan ee wax soo saarka antibody-ga laba-geesoodka ah waxaa ka mid ah isku-xidhka kiimikada, afar-ilmood oo hybridoma ah, iyo diyaarinta antibody-ga injineernimada hidde-sideyaasha. Kuwaas waxaa ka mid ah, habka isku-xidhka kiimikada wuxuu isku xiraa laba qaybood oo IgG ah ama laba F (ab ') ah oo aan waxba tarayn oo ah antibody-yo laba-geesood ah oo isku-dhafan ... Marka la samaynayo naqshadaynta antibody-ga laba-geesoodka ah, mabda'a falcelinta iskutallaabta antibody-ga ayaa la isticmaali karaa, laakiin falcelinta iskutallaabta antibody-ga waxay sababi kartaa falcelin aan gaar ahayn, sidaa darteed si taxaddar leh ayaa looga fiirsan doonaa isticmaalka wax ku oolka ah ee naqshadeynta antibody-ga laba-geesoodka ah, sida daaweynta antibody-ga laba-geesoodka ah.

Nadiifinta Unugyada Bispecific

Nadiifinta unugyada difaaca jirka ee bispecific waa habka lagu kala sooco laguna nadiifiyo unugyada difaaca jirka ee nadiifka sare leh. Laba hab, centrifugation iyo shaandhaynta qoto dheer, ayaa loo isticmaalaa in lagu saaro qaar ka mid ah wasakhda milmi karta. Unugga difaaca jirka ee bispecific-ga ah ee bartilmaameedka ah waxaa marka hore lagu qabtaa koromagrafiga isku-dhafka ah. Unugga difaaca jirka ee bispecific-ga ah ee IgG, borootiin A ayaa la isticmaalaa, halka unugyada difaaca jirka ee aan IgG ahayn sida bispecific-ga, koromagrafiga isku-dhafka ah ee ku salaysan silsiladda iftiinka ayaa la isticmaali karaa. Ka dib, antibody-ga waxaa lagu dhex geliyaa xaalado pH hooseeya muddo cayiman, taasoo keenta in qaab-dhismeedka borotiinka ee dusha sare ee baqshadda fayraska uu khalkhal galo, taasoo luma awoodda uu ku qaadi karo unugyada. Shaandhaynta qoto dheer ee dhexdhexaadka ah ayaa la sameeyaa si looga saaro wasakhda sida borotiinada unugyada martida loo yahay (HCP). Nadiifnimada unugyada difaaca jirka waxaa lagu hagaajiyaa hababka sida koromagrafiga is-weydaarsiga ion, fayras kasta oo harayna waxaa laga saaraa farsamooyinka nanofiltration ama ultrafiltration. Ugu dambeyntii, muunadda waa la ururiyaa waxaana lagu beddelaa kayd ku habboon.

Hab-socodka Shaqada ee Adeegga Wax-soo-saarka Antibodies-ka Bispecific

Tallaabooyinka	Macluumaadka Adeegga	Jadwalka
Isku-darka Hidda-wadaha	Soo saarista ilaa 3 qaab-dhismeed oo kala duwan oo taxane ah, oo ah soo saarista cusub ee plasmids-ka DNA-da ee unugyada difaaca jirka ee dib-u-habaynta ah	2-3 toddobaad
Tijaabada Miisaanka Yar	Soo saarista borotiinada unugyada difaaca jirka ee laba-jibbaaran → Muujin yar oo ku jirta khadadka unugyada naasleyda → Xaqiijinta SDS-PAGE → Falanqaynta isku xidhka antigens-ka borotiinka dib-u-habaynta ee ELISA → Isbarbardhigga unugyada iyo antibody-yada waalidka	5-6 toddobaad
Aqoonso	Muujinta labada unug ee ugu horreeya ee unugyada difaaca jirka ee dhererka buuxa leh → Falanqaynta isku xidhka ee ELISA → Qiimaynta ELISA ee unugyada difaaca jirka ee ay aqoonsadeen unugyada difaaca jirka ee ka hortagga jiirka	1 toddobaad
Soo saarista Unugyada Ka-hortagga	Wax soo saarka antibody-ga oo baaxad weyn leh	3-4 toddobaad

Faa'iidooyinkayaga

Waxaan daboolnaa adeegyada kor iyo hoos ee wax soo saarka antibody-ga, waxaana bixin karnaa adeegyo farsamo oo ka socda diyaarinta antibody-ga, nadiifinta antibody-ga laba-geesoodka ah iyo kala-soocidda iyo nadiifinta antibody-ga, taxanaha antibody-ga, xaqiijinta antibody-ga, iwm., kuwaas oo loo isticmaali karo daaweynta antibody-ga laba-geesoodka ah.

Muujinta Sare

Muujinta borotiinka sare ee unugyada CHO.
Kaydinta Xasilloon

> 2 toddobaad oo seeram ah heerkul ah 37 °C.
Ma jiro arrin Xal ah

>30 mg/ml
Lne-yada Unugyada Xasilloon

Khadad badan oo unugyo deggan oo leh antigens buro ah ayaa la sameeyay si loo xaqiijiyo antibodies badan oo laba-geesood ah