Sabis na Haɓaka Magungunan Hana Cututtukan Monoclonal

Kamfanin Alpha Lifetech Inc.zai iya samar da hasashen epitope, hada peptide, bayyanar furotin, samar da kwayoyin halittar polyclonal, ko monoclonal da kuma tsarkake kwayoyin halittar daga al'adar tantanin halitta, ruwan ascites ko kuma cikakken sinadarin serum.

Kamfanin Alpha Lifetech Inc.yana da ƙwarewa wajen samar wa abokan ciniki ayyuka daban-daban na layin salula masu karko, ciki har da ƙwayoyin hybridoma, layukan ƙwayoyin halitta masu gyara kwayoyin halitta, layukan ƙwayoyin cuta masu ƙwanƙwasawa, da layukan ƙwayoyin halitta masu yawan bayyanawa. Muna ƙoƙarin bayar da ayyukan layin salula na musamman na musamman ga kowane abokin ciniki kuma ayyukanmu an tabbatar da su kuma an amince da su.

Tsarin Ci Gaban Maganin Cututtukan Jiki

1. Tsarin Antigen da shirye-shirye
Waɗannan antigens na iya zama sunadarai, peptides, ko wasu ƙwayoyin halitta. Tsarin immunogens yana buƙatar cikakken tsari wanda ya haɗa da ilimin tsarin antigen, ilimin rigakafi, da tsarin bayarwa. Alpha Lifetech yana da fasahar ƙirar antigen wanda zai iya tsara immunogens ga abokan ciniki da ke buƙata.
Alpha Lifetech tana samar da samfuran furotin masu sake haɗawa ga abokan ciniki don zaɓar daga ciki. Muna da nau'ikan tsarin bayyanar furotin iri-iri, kamar tsarin bayyanar E. coli, tsarin bayyanar yeast, tsarin bayyanar kwari na baculovirus, tsarin bayyanar dabbobi masu shayarwa, da sauransu, don samar da furotin masu sake haɗawa da kuke buƙata don bincike.

2. Rigakafin dabbobi
Kamfanin Alpha Lifetech zai iya samar da garkuwar jiki ga abokan ciniki, kuma ana samun dabbobin garkuwar jiki kamar beraye, zomaye, tumaki, alpaca, aladu na Guinea, hamsters, da sauransu don ku zaɓa daga ciki.

3. Hanyoyin samar da rigakafi na monoclonal

●Samar da Mabs ta hanyar fasahar Hybridoma
Hybridoma wata ƙwayar halitta ce ta haɗin gwiwa da aka samar ta hanyar haɗa nau'ikan ƙwayoyin halitta guda biyu, wato layin ƙwayoyin halitta na linzamin kwamfuta da ƙwayoyin jini (lymphocyte B), inda ƙwayar halitta mai heterozygous za ta iya samar da ƙwayoyin rigakafi masu ci gaba da yaƙi da antigen of interest in vitro. Samar da layin ƙwayoyin halitta na hybridoma ya ƙunshi sassa uku: ƙirar antigen (haptens, ƙananan ƙwayoyin halitta, da peptides), rigakafin dabbobi, haɗa ƙwayoyin halitta, da kuma tantancewar clone mai kyau. Ta amfani da hanyoyin rigakafi na musamman, muna samun ingantaccen haɗin ƙwayoyin halitta (hybridomas 1-10 a kowace ƙwayoyin B dubu) da kuma yawan ƙwayoyin rigakafi, wanda ke ƙara yawan nasararmu a gwaje-gwajen nazari na gaba waɗanda suka haɗa da tantancewar sinadarai, gwaje-gwajen da aka yi a cikin ƙwayoyin halitta, da nazarin dabbobi.

●Samar da Mabs ta hanyar fasahar nuni ta phage
Nunin Phage yana ba da wata hanya don samar da ƙwayoyin rigakafi na monoclonal. Ana ware ƙwayoyin B-lymphocytes daga jinin dabbobi kuma ana cire mRNA ɗinsu. Ta hanyar PCR, ana canza wannan mRNA zuwa cDNA, wanda ke haɓaka dukkan sassan VH da VL. Daga nan ana haɗa waɗannan sassan zuwa vector, yawanci azaman gutsuttsuran masu canzawa guda ɗaya (scFv), tare da furotin na PIII na bacteriophage. Daga nan, ana amfani da wannan ginin don kamuwa da E. coli, wanda ke haifar da ƙirƙirar ɗakin karatu wanda ke ɗauke da kusan ƙwayoyin 10^10 ta hanyar allurar rigakafi tare da taimakon phage. Daga nan E. coli zai iya fitar da ƙwayoyin cuta da ke ɗauke da sassan VH da VL a matsayin wani ɓangare na gashinsa. Bayan haka, ana iya zaɓar takamaiman sassan VH da VL waɗanda ke niyya ga antigen ɗin da ake sha'awa kuma ana amfani da su don sake haɗa E. coli tare da bacteriophage. Sannan ana ware ƙwayoyin da ke ɗauke da plasmid kuma a jera su.

4. Fusion screening

●Haɗawar Kwayar Halitta da Zaɓi
Sannan a haɗa ƙwayoyin B da aka girbe tare da ƙwayoyin myeloma, waɗanda ƙwayoyin cutar kansa ne waɗanda za su iya yaɗuwa har abada a cikin al'ada. Yawanci ana samun haɗakar ta amfani da dabara kamar haɗakar polyethylene glycol (PEG).
Bayan haɗuwa, ana shuka ƙwayoyin hybridoma a cikin wani zaɓi wanda zai ba hybridomas kawai damar rayuwa. Maganin yawanci yana ɗauke da aminopterin, wanda ke hana haɓakar ƙwayoyin myeloma marasa haɗuwa.

●Nunawa da Rufewa
Ana tantance ƙwayoyin hybridoma da suka samo asali don samar da ƙwayoyin rigakafi waɗanda suka dace da antigen ɗin da aka yi niyya. Yawanci ana yin wannan ta amfani da dabaru kamar enzyme-linked immunosorbent assay (ELISA), Western blot, immunoprecipitation, da flow cytometry.

Da zarar an gano ƙwayoyin hybridoma da ke samar da antibody, ana yin kwafi don samar da yawan ƙwayoyin halitta iri ɗaya. Wannan yana tabbatar da cewa antibody da kowace ƙwayar hybridoma ke samarwa iri ɗaya ne.

5. Tabbatar da Aikin Antibody
Alpha Lifetech tana ba da tabbacin aikin rigakafi, kamar Western blotting, immunohistochemistry, ko gwaje-gwajen aiki, don tantance ƙwayoyin rigakafi na monoclonal da kuma tabbatar da takamaiman ƙwayoyin rigakafi, kusanci, da aiki.

6. Inganta garkuwar jiki
Tsarkakewar ƙwayoyin rigakafi daga sinadarin halitta ta amfani da dabaru kamar furotin A ko furotin G affinity chromatography, da kuma Alpha Lifetech suma na iya samar da hanyoyin gyara ƙwayoyin rigakafi don inganta haɗin gwiwar ƙwayoyin rigakafi.

7. Samar da maganin hana garkuwar jiki
Alpha Lifetech na iya tantance ƙwayoyin rigakafi don aikace-aikacen tantancewa mai yawa da kuma samar da babban adadi.

tambayoyin da ake yawan yi akai-akaitambayoyin da ake yawan yi akai-akai

Tambayoyi da Amsoshi game da ci gaban ƙwayoyin cuta na monoclonal

1

Yadda ake zaɓar hybridoma mai dacewa?

A lokacin haɗakarwa, ana shuka faranti masu rijiyoyi 10-15 masu girman 96 tare da cakuda haɗakar haɗakar. Ana ciyar da kowace faranti ta hanyar zaɓin HAT-IMDM kuma a adana ta a cikin injin haɗakar carbon dioxide a digiri 37 na Celsius. Kwanaki 9-14 bayan haɗuwa, ana gwada supernatants masu launin shuɗi don ganin kasancewar takamaiman antibody mai ban sha'awa.
2

Yadda za a inganta ingancin zaɓi?

Haɗa ƙwayoyin plasma da takwarorinsu na myeloma ba shi da tasiri 100%. Ko da a cikin yanayi mafi kyau da kuma mafi kyawun motsa jiki, haɗa ƙwayoyin har yanzu yana samar da cakuda ƙwayoyin da ba su da haɗuwa da haɗuwa waɗanda ke buƙatar a raba su. Don inganta ingancin tsarin zaɓi, ƙwayoyin myeloma da ake amfani da su don haɗuwa ba su da HGPRT, wani babban enzyme a cikin hanyar ceto ta nucleotide. Daga nan aka haɓaka haɗin a cikin wani matsakaici na HAT, inda ƙwayoyin da ke da enzyme HGPRT kawai, hybridomas waɗanda suka gaji HGPRT daga ƙwayoyin plasma, suke rayuwa.
3

Yadda ake tantance layukan ƙwayoyin hybridoma don aikin antibody?

Kimanta nau'ikan hybrid wani muhimmin mataki ne. Yawanci, ana samar da gwajin hybridoma, clone, da subclone supernatant ta hanyar enzyme-linked immunosorbent assay (ELISA), Western blot assay, da fluorescence activated cell sorting (FACS). Za mu zaɓi mu gudanar da duk binciken supernatant a dakin gwaje-gwajenmu.
4

Ba da cikakken bayani kan yadda ake gudanar da gwajin ayyukan rigakafi?

Hybridoma supernates da za a gwada na iya kasancewa daga samfura 500 zuwa 1,440 kuma za su kasance a shirye don gwaji kafin kwanaki 9-14. Za mu iya tasowa cikin kwanaki uku zuwa biyar ko kuma mu kasance a shirye a rana ɗaya, don haka yana da mahimmanci ga dakin gwaje-gwajen abokin ciniki ya shirya makonni kafin lokaci tare da takamaiman gwaje-gwajen tantancewa na biyu da aka zaɓa.
5

Me yasa ake buƙatar a yi wa hybridomas masu kyau tiyata?

Bayan an gano Wells masu kyau a lokacin aikin tantancewa na farko na ELISA, an canza hybridomas zuwa babban girma, watau, faranti 24-rijiya, don shirya aikin subcloning. Yawanci ana yin subcloning na hybridomas ta hanyar iyakanceccen hanyar dilution don tabbatar da ware monoclones masu karko. Dabarar ta ƙunshi dilution na al'adun hybridoma da kuma watsa su cikin faranti 96-rijiya don cimma monoclonality (ƙwayar halitta ɗaya a kowace rijiya). Ya kamata a yi aƙalla dilution guda biyu masu iyaka don rage haɗarin kamuwa da gaurayen hybridoma.
6

Menene fa'idodin amfani da fasahar hybridoma don gano ƙwayoyin cuta?

An yaba wa layukan ƙwayoyin halitta masu haɗaka saboda iyawarsu ta samar da ƙwayoyin rigakafi masu ƙarfi, kwanciyar hankali, da takamaiman abubuwa ta hanyar da ta fi araha.