Iqonga loPhuhliso lwe-Antibody Display yePhage

I-Alpha Lifetech, enamava ayo amakhulu kwiprojekthi yokufumanisa i-antibody, inokubonelela ngeenkonzo ze-antibody ze-monoclonal kunye ne-polyclonal ezenzelwe wena kwiintlobo ngeentlobo, kunye neenkonzo zokwakha kunye nokuhlola ithala leencwadi le-antibody. I-Alpha Lifetech inokubonelela abathengi ngemveliso ye-antibody eqinisekisiweyo kunye neemveliso ze-antibody eziphinda zihlanganiswe, kunye neenkonzo ezifanayo, ukulungiselela ii-antibodies ezisebenzayo, ezichanekileyo kakhulu, nezizinzileyo. Ngokusebenzisa amaqonga e-antibody apheleleyo kunye nobunjineli be-antibody, sinikezela ngeenkonzo ezigubungela imveliso ye-antibody ephezulu nesezantsi, kubandakanya iinkonzo zobugcisa ezifana nokulungiselela i-antibody, ukucoca, ukulandelelana kwe-antibody, kunye nokuqinisekiswa.

I-Alpha Lifetech ineqonga lokufumana i-antibody evuthiweyo ebonelela abathengi ngeenkonzo ezahlukeneyo eziquka ukukhuselwa kwezilwanyana, ukwakhiwa kunye nokuhlolwa kwethala leencwadi le-antibody ye-phage display, ukulandelelana kwe-antibody, ukubonakaliswa kwe-antibody, ukucocwa kwe-antibody, ukuqinisekiswa kwe-antibody, kunye nokubhalwa kwe-antibody ngokusekelwe kwitekhnoloji ye-hybridoma, itekhnoloji ye-single B cell, itekhnoloji ye-phage display, nokunye. I-Alpha Lifetech inomgca wemveliso oqhelekileyo, kwaye ukongeza kwiinkonzo zokulungiselela i-antibody, iinkonzo ezixhasayo ezifana nokwenziwa kwe-antibody humanization, ukuvuthwa kwe-antibody affinity, uyilo kunye nophuhliso lwamayeza e-ADC, kunye noyilo lwe-CAR-T olulandelayo lunokubonelelwa. Kwangaxeshanye, i-Alpha Lifetech yakhe ithala leencwadi le-antibody ngokusekelwe kumaqonga etekhnoloji ye-M13, T4, kunye ne-T7 phage display, enomthamo wokugcina ukuya kuthi ga kwi-10^8-10^9. Isantya esihle, izinga lokufakwa, kunye nokwahluka kwethala leencwadi konke kunokufikelela ngaphezulu kwe-90%.

Itekhnoloji yokubonisa iiPhage

Ukuveliswa kwee-antibodies ze-monoclonal kwaqalwa ngokusetyenziswa iteknoloji ye-hybridoma monoclonal antibody ukuvelisa i-mouse hybridoma monoclonal antibody. Ngokukodwa, ngokudibanisa iiseli ze-spleen ezivela kwiimpuku ezigonyiweyo kunye neeseli ze-myeloma ezivela ebantwini okanye kwiimpuku, kwakheka iiseli ze-hybridoma, ezikhupha ii-antibodies ezithile ze-hybridoma monoclonal antibody. Ngokuguqulwa kwee-antibodies ze-monoclonal zempuku ezenziwe ngabantu kunye nokuguqulwa kobunjineli bemfuza bee-antibodies ukuze zinike indawo ehlala ihleli ye-immunoglobulin yomntu, ukuqina kwazo kwe-immunogenicity kunokuncipha. Ubunjineli be-monoclonal antibody busetyenziswa kakhulu kwizicelo ze-biomedical kunye neklinikhi.

I-Alpha Lifetech inokubonelela ngeenkonzo zobunjineli be-antibody enye ukusuka kuyilo lweprotocol yophando lwangaphambi kweklinikhi ukuya ekufumaneni i-antibody drug conjugates (ADCs) kunye nokuqinisekiswa kwezilwanyana. I-antibody drug conjugates (ADCs) inokuhambisa amayeza e-chemotherapy kwiiseli zomhlaza. Emva kokubophelela kwiithagethi ezithile ezichazwe kwiiseli zomhlaza, i-ADC ikhupha amayeza e-cytotoxic kwiiseli zomhlaza. I-Alpha Lifetech inokubonelela abathengi ngeenkonzo ezipheleleyo zokuvuthwa kwe-antibody affinity. Ngokulungiswa kwe-mutation okuphambili kunye netekhnoloji yokujonga i-phage ephezulu, ii-antibodies ezine-affinity ethile ziyahlolwa kuqala, kwaye emva koko kuqaliswa ii-amino acid mutations ukuvelisa iintlobo ezahlukeneyo. Emva koko, ii-antibodies ezine-affinity ephezulu zahlolwa kwaye zahlolwa kusetyenziswa iindlela zokuhlola. Emva kwemijikelo emininzi yokwenza ngcono kunye nohlalutyo lwesakhiwo, ii-antibodies ezine-affinity ephezulu kunye neenkcukacha eziqinileyo ekugqibeleni zafunyanwa.

I-Alpha Lifetech inokwakha iintlobo ezahlukeneyo zeelayibrari zokubonisa i-antibody phage, kubandakanya iilayibrari ze-immune, iilayibrari zemveli, iilayibrari ze-semi synthetic, kunye neelayibrari ze-synthetic. Ngokusekelwe kumthamo ophezulu weelayibrari ze-antibody, ii-antibodies ezithile kakhulu zinokufunyanwa. Iivektha ze-phagemid ezininzi ezifana ne-pMECS, i-pComb3X, kunye ne-pCANTAB 5E zinokubonelelwa, kunye neentlobo ezifana ne-TG1 E. coli, i-XL1 Blue, kunye ne-ER2738. Ukongeza kumthamo welayibrari ukuya kuthi ga kwi-10^9, izinga lokufakwa kweqhekeza elijoliswe kulo lelayibrari nalo liphezulu, okudala iimeko ezaneleyo zokuhlola ii-antibodies ezanelisa abathengi. Itshathi yokuhamba kwenkonzo yokulungiselela i-antibody ye-Alpha Lifetech esekelwe kubuchwepheshe bokubonisa i-phage iboniswe kuMfanekiso 1.

Umzobo 1 Umzobo wenkqubo yokulungiselela ii-antibodies ze-monoclonal ezisekelwe kwitekhnoloji yokubonisa i-phage.

Ukuhamba komsebenzi wokuvelisa i-Antibody Display yePhage Display

Amanyathelo	Umxholo weNkonzo	Umgca wexesha
Inyathelo 1: Ukugonywa kwezilwanyana	(1) Ukugonywa kwezilwanyana izihlandlo ezi-4, ukugonywa kwe-booster idosi e-1, iyonke iidosi ezi-5 zigonyiwe. (2) I-serum engalunganga ngaphambi kokuba kugonywe, kwaye i-ELISA yenziwe kwidosi yesine ukuze kufunyanwe i-serum titer. (3) Ukuba i-serum antibody titer yedosi yesine iyahlangabezana neemfuno, idosi enye eyongezelelweyo yokugonywa iya kunikwa kwiintsuku ezi-7 ngaphambi kokuqokelelwa kwegazi. Ukuba ayihlangabezani neemfuno, ukugonywa rhoqo kuya kuqhubeka. (4) Amandla afanelekileyo, ukuqokelelwa kwegazi kunye nokwahlukana kwee-monocytes	Iiveki ezili-10
Inyathelo lesi-2: Ukulungiswa kwe-cDNA	(1) I-PBMC Total RNA Extraction (Ikhithi Yokukhupha i-RNA) (2) Ukulungiswa kwe-RT-PCR ethembekileyo kakhulu ye-cDNA (ikhithi yokubhala umva)	Usuku olu-1
Inyathelo lesi-3: Ukwakhiwa kweThala leeNcwadi le-Antibody	(1) Ukusebenzisa i-cDNA njengetemplate, ii-genes zandiswa ngemijikelezo emibini ye-PCR. (2) Ukwakhiwa kwePhage kunye nokuguqulwa: i-gene splicing phagemid vector, ukuguqulwa kwe-electroporation yebhaktiriya ye-TG1 host, ukwakhiwa kwethala leencwadi le-antibody. (3) Ukuchonga: Khetha ngokungacwangciswanga ii-clones ezingama-24, izinga lokuchongwa kwe-PCR kunye nezinga lokufakwa. (4) Ukulungiswa kwe-phage encediswayo: Ukwandiswa kwe-phage ye-M13 + ukucoca. (5) Uhlangulo lwethala leencwadi lokubonisa iiPhage	Iiveki ezi-3-4
Inyathelo lesi-4: Uvavanyo lweThala leeNcwadi lwe-Antibody (imijikelo emi-3)	(1) Uvavanyo oluqhelekileyo lwemijikelo emi-3 (uvavanyo lwesigaba esiqinileyo): uvavanyo loxinzelelo lokususa ii-antibodies ezingezizo ezithile kangangoko kunokwenzeka. (2) I-bacteriophage ekhethiweyo yokukhulisa i-clone enye+ukubonakaliswa okubangelwe yi-IPTG+ukufunyanwa kwe-ELISA yee-clone ezilungileyo. (3) Zonke ii-clones ezilungileyo zikhethwe ukuze kulandelelwe iijini.	Iiveki ezi-4-5

Inzuzo yeTekhnoloji yoBoniso lwePhage

I-Alpha Lifetech inamava amaninzi kwicandelo lophuhliso lwee-antibody. Kwiminyaka edlulileyo, i-Alpha Lifetech iseke iqonga elibanzi lophuhliso lwee-antibody.

Iinkonzo zeNkxaso

Singabonelela ngeenkonzo ezahlukeneyo zokwakha ithala leencwadi lokuzikhusela komzimba ezisekelwe kwizilwanyana kunye neenkonzo zokuhlola ii-antibody zendalo ngokweemfuno zabathengi.

Iithagethi ezininzi

Iinkonzo ezininzi zokufumanisa ii-antibody ezijoliswe kuzo ziyafumaneka: iiproteni, iipeptide, iimolekyuli ezincinci, iintsholongwane, iiproteni ze-membrane, i-mRNA, njl.njl.

IiVektha ezininzi

Inkonzo yokwakha ilayibrari eyenzelwe wena, singabonelela ngeevektha zebhakteriophage ezahlukeneyo kuquka i-PMECS, i-pComb3X, kunye ne-pCANTAB 5E, kwaye siziguqule ngokweemfuno zabathengi.

Iqonga labantu abadala

Umthamo wokugcina unokufikelela kwi-10 ^ 8-10 ^ 9, amazinga okufakwa angaphezulu kwe-90%, kwaye ukuqina kwee-antibodies ezifunyenwe ngokuhlolwa ngokubanzi kukwinqanaba le-nM pM.

INKONZO EFANELEKILEYO

Amaqhinga amaninzi okuphuhlisa i-Antibody

Inkonzo yoPhuhliso lwee-Antibody zeMonoclonal

Iqonga leTekhnoloji yeHybridoma

Iqonga lokuHlela iiSeli ze-B enye

Iqonga loPhuhliso lwe-Antibody Display yePhage

uphuhliso lwe-antibody ye-monoclonal-Alpha Lifetech

Inkonzo yoPhuhliso lwee-Antibody zeMonoclonal

Singabonelela ngeenkonzo zophuhliso lwee-antibody ze-monoclonal ezikumgangatho ophezulu, ezicocekileyo kakhulu, nezicacileyo kakhulu, kubandakanya ukuveliswa kwee-antibodies ze-monoclonal zegundane kunye nee-antibodies ze-rabbit monoclonal.

Iqonga leTekhnoloji yeHybridoma

Kubandakanya inkqubo yokugonya, iinkonzo zokulungiselela i-antibody, ukucocwa kwe-antibody, ukulandelelana kwe-antibody ephezulu, ukuqinisekiswa kwe-antibody, njl.

Iqonga lokuHlela iiSeli ze-B enye

I-Alpha Lifetech ineengenelo ngexesha lokuhlola kunye nokufumana ii-antibodies ezisemgangathweni ophezulu. Ingabonelela ngoyilo lwe-antigen, ukwenziwa, kunye nokuguqulwa, ukukhuselwa kwezilwanyana, ukuhlolwa kokutyetyiswa kweseli enye ye-B, kunye nokulandelelana kweseli enye.