Pulatifomu ya Ukadaulo wa Hybridoma
Alpha Lifetech imapatsa makasitomala zinthu ndi ntchito zabwino kwambiri zotsimikizira kuti ma antibodies ndi ma antibodies opangidwanso.
Pemphani Mtengo01
Pulatifomu ya Ukadaulo wa Hybridoma
Alpha Lifetech ili ndi chidziwitso chochuluka pakupanga ma antibodies. Kwa zaka zambiri, Alpha Lifetech yakhazikitsa njira yonse yopangira ma antibodies (kuphatikizapo kupanga ma antibodies a monoclonal, kupanga ma antibodies a polyclonal, kupanga ma antibodies a monoclonal recombinant, ndi ntchito zina zokonzekera ndi kupanga ma antibodies).
Alpha Lifetech imapatsa makasitomala zinthu zabwino kwambiri zoteteza ma antibodies ndi ma antibodies opangidwanso, ndipo ma antibodies okonzedwawo ali ndi mawonekedwe apadera komanso kukhazikika bwino. Zipangizo zambiri zoyeretsera ma antibodies ndi zida zingagwiritsidwe ntchito popereka ntchito zoyeretsa ma antibodies kuchokera kuzinthu zosiyanasiyana monga kalulu, nkhosa, nkhuku, ma antibodies a monoclonal a mbewa, komanso ntchito zoyeretsa ma protein A/G affinity ndi ntchito zolekanitsa ma antibodies.
Kutengera njira zosiyanasiyana zopangira ma antibodies monga ukadaulo wa maselo a hybridoma, Alpha Lifetech ikhoza kupereka ntchito zomwe zakonzedwa kuti zigwiritsidwe ntchito pa ma antibodies a monoclonal ndi polyclonal amitundu yosiyanasiyana. Kuphatikiza apo, ntchito zosiyanasiyana kuyambira pakutsata njira yopezera ma antibodies ambiri, kukonzekera ma antibodies, kuyeretsa ma antibodies mpaka kutsimikizira ma antibodies zitha kuperekedwanso. Tidzasankha njira yoyeretsera kutengera cholinga chenicheni cha kasitomala. Kutengera njira zonse za nsanja monga nsanja yopangira ma antibodies ndi nsanja ya mapuloteni, Alpha Lifetech imapereka ntchito zopanga ma antibodies kumtunda ndi kumunsi, kupereka kuyeretsa ma antibodies, kutsata njira yopezera ma antibodies ambiri, kutsimikizira ma antibodies ndi ntchito zina zaukadaulo.
Ukadaulo wa Hybridoma
Ukadaulo wa Hybridoma ungagwiritsidwe ntchito pokonzekera ma antibodies a monoclonal. Mwa kuphatikiza maselo a spleen kuchokera ku mbewa zomwe zapatsidwa katemera ndi maselo a myeloma ochokera kwa anthu kapena mbewa, maselo a hybridoma amapangidwa ndikupereka ma antibodies enieni. Ma antibodies a monoclonal opangidwa ndi mbewa angagwiritsidwe ntchito mu kafukufuku wasayansi monga enzyme-linked immunosorbent assay, radioimmunoassay, immunohistochemistry, flow cytometry, protein purification, ndi zina zotero. Zitsanzo za ma antibodies a monoclonal zimaphatikizapo ascites, hybridoma supernatant, ndi zina zotero. Chitsanzo chomwe chimapezeka poika ma cell a hybridoma m'mimba mwa nyama ndikugwiritsa ntchito malo ozungulira m'mimba mwa nyama kuti apange ma antibodies chimatchedwa ascites sample. Ma cell a Hybridoma omwe amalowetsedwa m'mimba mwa nyama amatha kufalikira ndikutulutsa ma antibodies a monoclonal, omwe amatha kudziunjikira mu ascites. Chifukwa cha kuthekera kwa zodetsa (mapuloteni a nyama, mahomoni, ndi zina zotero) m'ma antibodies awa, kuyeretsa kumafunika. Kutengera ntchito yotulutsa ma B lymphocytes, ma antibodies a monoclonal nthawi zambiri amatulutsidwa mu malo osungiramo zinthu akamakulitsa ma cell a hybridoma. Madzi okhala ndi ma antibodies a monoclonal mu culture medium amatchedwa hybridoma supernatant. Kuyera kwa ma antibodies a monoclonal mu supernatant ndi kwakukulu, koma kuyeretsa kwina kumafunika kuti pakhale kukonzekera kwakukulu chifukwa amapezeka kudzera mu in vitro culture.
Kuyeretsa ma antibodies kumatha kusiyanitsa ma antibodies a high-purity kuchokera ku ma complexes. Pakadali pano, njira zoyeretsera ma antibodies ndi kulekanitsa ndi kuyeretsa ma antibodies zimagwiritsidwa ntchito kwambiri, makamaka kuphatikiza kuyeretsa mapuloteni A/G, ion exchange chromatography, kusefa kwa gel, precipitation ndi hydrophobic interaction chromatography. Pakati pawo, ntchito yoyeretsa ma protein A/G affinity ndi yotchuka kwambiri m'magwiritsidwe osiyanasiyana oyeretsa ma antibodies. Njira ya Alpha Lifetech yokonzekera ma antibodies a monoclonal kutengera ukadaulo wa hybridoma ikuwonetsedwa mu Chithunzi 1.
Chithunzi 1 Chithunzi cha njira yokonzekera ma antibodies a monoclonal kutengera ukadaulo wa hybridoma.
Ukadaulo wa Mankhwala Oletsa Kuphatikizika kwa Antibody ndi Hybridoma
Mankhwala oletsa kutupa (ADC) ndi mankhwala atsopano achilengedwe omwe amaphatikiza ma antibodies ndi mankhwala oletsa kutupa. Mankhwala oletsa kutupa amaphatikiza mphamvu yeniyeni yozindikira ma antibodies ndi mphamvu yamphamvu yopha mankhwala oletsa kutupa, ndipo amapereka mankhwala oletsa kutupa m'maselo a chotupa kudzera mu ma antibodies, motero amapeza chithandizo choyenera. Gawo la ma antibodies mu mankhwala a ADC nthawi zambiri limakonzedwa kudzera muukadaulo wa hybridoma kapena njira zina zopangira ma antibodies, monga ukadaulo wa majini oletsa kutupa. Ukadaulo wa Hybridoma umapereka zinthu zopangira ma antibodies apamwamba kwambiri a mankhwala a ADC. Mankhwala a ADC ndi ma antibodies a monoclonal okonzedwa kudzera muukadaulo wa hybridoma amachita ntchito zofunika kwambiri m'magawo monga chithandizo cha khansa. Angagwiritsidwe ntchito okha kapena kuphatikiza kuti awonjezere mphamvu ya chithandizo.
Ma antibodies a monoclonal okonzedwa ndi ukadaulo wa hybridoma ali ndi mawonekedwe apadera, amatha kumangirira ndikuzindikira ma antigen enieni molondola, ndipo amatha kupangidwa pamlingo waukulu. Kudzera mu ukadaulo wa hybridoma sequencing, majini a antibody m'maselo a hybridoma amatha kuchotsedwa ndikusanjidwa kuti apeze ma sequence ofanana, zomwe zimathandiza pakutulutsa ma antibodies pambuyo pake. Ma antibodies a monoclonal okonzedwa kudzera mu ukadaulo wa hybridoma angagwiritsidwe ntchito kuzindikira zizindikiro za matenda enaake, monga zizindikiro za chotupa, ma antigen a mavairasi, ndi zina zotero, kupereka chithandizo champhamvu pakuzindikira matenda koyambirira. Kuphatikiza apo, ingagwiritsidwenso ntchito mu zoyeserera monga immunoblotting ndi immunohistochemistry kuti iphunzire mawonekedwe a mapuloteni, malo, kapangidwe, ndi ntchito.
Ukadaulo wa hybridoma Kupanga Antibody Service Workflow
| Masitepe | Zamkati mwa Utumiki | Mndandanda wa Nthawi |
|---|---|---|
| Gawo 1: Katemera wa ziweto | (1) Katemera wa ziweto nthawi 4, katemera wowonjezera mlingo umodzi, mlingo wonse wa 5 wa katemera. (2) Seramu yoipa isanatengedwe katemera, ndipo ELISA idachitidwa pa mlingo wachinayi kuti ipeze titer ya seramu. (3) Ngati mlingo wa antibody wa seramu wa mlingo wachinayi ukukwaniritsa zofunikira, mlingo umodzi wowonjezera wa katemera udzaperekedwa masiku 7 magazi asanatengedwe. Ngati sukwaniritsa zofunikira, katemera wanthawi zonse udzapitirira. (4) Mphamvu yoyenerera, maselo a spleen amalekanitsidwa kuti maselo agwirizane. | Masabata 10-12 |
| Gawo 2: Kusakaniza ndi Kuwunika | (1) Kusakaniza: Pambuyo pa katemera, maselo a mbewa a ndulu anasonkhanitsidwa ndikuphatikizidwa ndi maselo a myeloma. (2) Kuchulukana ndi kuyezetsa: Maselo a hybridoma omwe adapezeka adachulukitsidwa ndi kuyezetsa ELISA kuti apeze ma clones ogwira ntchito bwino kwambiri. | Masabata 6 |
| Gawo 3: Subclone | Kuchotsa maselo a mayi abwino 15-25 pogwiritsa ntchito njira yochepa yochepetsera, kenako n’kupeza maselo abwino 10-15 | Masabata 3-4 |
| Gawo 4: Kupanga Ascites ndi Kuyeretsa Ma Antibody | (1) Kupanga ma ascites: Mitundu 10-15 ya hybridoma imagwiritsidwa ntchito popanga ma ascites m'makoswe (ngati mitundu yambiri ya hybridoma ipezeka, idzatumizidwa mu mawonekedwe a selo) (2) Kuzindikira ma Ascites: Kuzindikira ma Ascites ELISA, mphamvu (puloteni/kachirombo>10 ^ 5, molekyulu yaying'ono/peptide>10 ^ 4) yoyeretsera pansi pa madzi (3) Kuyeretsa Mapuloteni A/G: Kuyeretsa Ma Ascites | Masabata 3-4 |
| Gawo 5: Kutumiza | (1) Perekani mitundu 3-5 ya ma antibodies a monoclonal hybridoma. (2) Selo iliyonse ya hybridoma imapereka 1mg ya antibody yoyeretsedwa (kuyeretsa ascites). (3) Lipoti la Njira Yoyesera | Sabata imodzi |
Mitundu Yambiri
Chitetezo chamthupi chingaperekedwe kwa nyama zosiyanasiyana, kuphatikizapo mbewa, akalulu, nkhosa, ndi zina zotero koma osati zokhazo.
Zolinga Zambiri
Kutengera ukadaulo wa hybridoma, ntchito zambiri zopezera ma antibodies zomwe zilipo: mapuloteni, ma peptide, mamolekyu ang'onoang'ono, ma virus, mapuloteni a membrane, mRNA, ndi zina zotero.
Zolemba Zolondola
Miyezo yowongolera khalidwe la QC (mphamvu ya chitetezo chamthupi, kuwongolera khalidwe la PBMC, kuwongolera khalidwe la laibulale, ndi kuwongolera khalidwe lotsimikizira), malipoti oyesera a Chingerezi, zolemba zoyambirira zoyesera
Pulogalamu Yopangidwira Munthu Aliyense
Kusintha kwa pulogalamu ya munthu mmodzi payekha kuti ikwaniritse zosowa za kafukufuku wa makasitomala osiyanasiyana
Utumiki Wopanga Ma Antibody a Monoclonal
Tikhoza kupereka ntchito zapamwamba kwambiri, zoyera kwambiri, komanso zapadera kwambiri zopanga ma antibodies a monoclonal, kuphatikizapo kupanga ma antibodies a monoclonal a mbewa ndi ma antibodies a kalulu a monoclonal.
Pulatifomu ya Ukadaulo wa Hybridoma
Kuphatikizapo pulogalamu ya katemera, ntchito zokonzekera ma antibodies, kuyeretsa ma antibodies, kutsata ma antibodies ambiri, kutsimikizira ma antibodies, ndi zina zotero.
Pulatifomu Yosankhira Maselo Awiri Awiri
Alpha Lifetech ili ndi ubwino pa nthawi yoyezetsa magazi ndi kupeza ma antibodies apamwamba kwambiri. Ikhoza kupereka kapangidwe ka ma antigen, kupanga, ndi kusintha, chitetezo cha nyama, kuyesa kuchuluka kwa maselo a B amodzi, ndi kutsata maselo amodzi.
Pulogalamu Yopangira Ma Antibody Yowonetsera Phage Display
Alpha Lifetech ikhoza kupereka ntchito zaukadaulo zopangira ma antibodies a phage display kuyambira kukonzekera ma antibodies, kuyeretsa ma antibodies, kutsata ma antibodies, ndi zina zotero.
Zida Zaukadaulo Zogwirizana
Chiyambi cha Ukadaulo wa HybridomaKANEMA
Ngati muli ndi mafunso aliwonse, chonde musazengereze kulankhulana nafe nthawi iliyonse.
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2018-07-16 
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