Serivisi yo Gushakisha Antibody ya VHH mu Gice Gitandukanye Nk'umutanga serivisi w'umwuga mu bijyanye no kwerekana no gukora antibody, Alpha Lifetech ishobora gutanga umusaruro wihuse w'amasomero ya antibody imwe.
Saba Igiciro
Serivisi yo kuvumbura ubudahangarwa bw'umubiri bwa VHH Alpha Lifetech Inc. ishobora gutanga umusaruro wihuse wa antibody imwe-domain (antibody ya VHH). Ikoranabuhanga ryacu ryo kwerekana phage ryemerera abahanga mu bya siyansi kugaragaza ibisabwa kugira ngo bagaragaze kandi bagaragaze imiterere y’uturemangingo.
Alpha Lifetech Ishobora gutanga Kubaka no gusuzuma isomero rya VHH Ubudahangarwa bw'umubiri Abahanga mu bya siyansi bo muri Alpha Lifetech Inc. bafite uburambe bwinshi mu kubaka no kugaragaza antibonda zishingiye ku rubuga rwacu rwa phage display. Isomero rya antibondari imwe rikingira rikwiriye gukora antibondari nyinshi za VHH cyangwa NAR V zihariye, bityo hirindwa imbaraga zo gukura kwa antibondari imwe mu buryo bwa vitro. Aha, dukora isomero rya antibondari imwe ridasanzwe rya antibondari ku bakiriya bacu riturutse ku binyabuzima bya alpaca, ingamiya, llamas cyangwa sharks, ndetse n'abantu (uturemangingo twa PBL dukoreshwa na antibondari muri vitro). Binyuze mu guhindura transcription na polymerase chain reaction, isomero rya antibondari imwe rigizwe na clones miliyoni 10-100 rikorwa buri gihe.
Kubaka no gusuzuma isomero rya VHH syntheic Amasomero y’uturemangingo tw’ubwoko bumwe akunze gutegurwa uhereye kuri CDR1 na CDR3 ya VHH cyangwa VNAR idafite ubunararibonye. Iyi somero y’uturemangingo tw’ubwoko bumwe mu buryo busanzwe ni isomero ridafite ubunararibonye kandi rifite imiterere myinshi ya VHH 3x10^10 ituma habaho ubwoko butandukanye bushimishije cyane. Ni ubunararibonye bityo ntisaba gukingirwa llama cyangwa shark. Ibi bidufasha kuzigama igihe kinini cyo kubona antibody runaka kandi bigatuma hashobora gutoranywa antibody ku poroteyine zitari iz’ubudahangarwa cyangwa izibitswe. Ayo masomero y’uturemangingo ni isoko nziza y’uturemangingo tw’ubwoko bumwe mu kurwanya antigens ubwayo, zitari iz’ubudahangarwa, kandi zifite uburozi kuko amasomero akenshi aba ari manini kandi atandukanye. Mu gihe guhitamo antibodys z’ubwoko bumwe zishongesha cyane ari ikibazo, twakoze sisitemu ya vector ya shuttle phagemid, harimo na phagemid nyinshi, zishobora kugaragaza antibodys z’ubwoko bumwe muri E. coli (umusemburo) cyangwa zikagaragara ku buso bw’uduce twa phage.
Imiterere Ubwoko Ingano y'Isomero
Naive scFv Umuntu 108 kugeza kuri 1010
Naive scFv Imbeba 108 kugeza kuri 1010
Naive scFv Urukwavu 108 kugeza kuri 1010
Kuki twahitamo Dushobora gutanga serivisi zirambuye zijyanye n’ibinyabuzima bito, harimo gukingira inyamaswa (harimo alpacas, ingamiya, nibindi), kubaka no gusuzuma isomero rya VHH, kugaragaza no kwemeza uturemangingo twa VHH, guhindura uturemangingo twa VHH nk’umuntu, nibindi.
Antibody ya VHH imwe-dome ni iki? Antibody imwe (sdAb), cyangwa nanobody, VHH, ni igice cy’ubwoko bw’uduce tw’uturemangingo tw’umubiri, tugaragaza antibody ntoya cyane yagaragaye ko ifite akamaro kanini. Ifite uburemere bwa molekile bwa kDa 12-15, antibody imwe, igizwe gusa n’ubwoko bumwe bw’uturemangingo tw’umubiri uremereye kandi ikaba injeniyeri ikomoka ku bwoko bumwe bw’uturemangingo tw’umubiri tw’udusimba twa camelids (VHH) cyangwa IgNAR (VNAR) tudatakaje ubushobozi bwo gufata antigen.
Antibodies zo mu bwoko bumwe zifite ibyiza byinshi: Ubufatanye bwinshi n’uturemangingo duto, imiterere myiza ya fizikike, no gukora ibintu byoroshye. Imiterere ya single-domain hamwe no koroshya gukoresha gene za single-domain antibody bituma ziba nziza mu gukura kwa affinity muri vitro.
Kugereranya hagati y’umuti urwanya VHH mu buryo bumwe n’umuti urwanya indwara usanzwe
Antibody y'idosiye imwe
Ubudahangarwa bw'umubiri busanzwe
Ingano
Itsinda rito, rigizwe n'uruhererekane rumwe ruremereye (V)H H)~13 kDa
Iminyururu minini, iremereye n'iyoroshye ~ 120-150 kDa
Ahantu hahurira Antigen
Monomeric VH Igice cya H gikenewe kugira ngo antigen ifatanye
Iminyururu yombi ikenewe kugira ngo antigen ifatanye kandi ihamye
Inzira yo hasi
Irakwiriye cyane mu buhanga bwo hasi
Ubuhanga buke cyane bitewe n'imiterere igoye
Ituze
Igumana ubusugire n'imbaraga ku rugero rukabije rwa pH n'ubushyuhe
Ntishobora kwihanganira pH cyangwa ubushyuhe bukabije
Uburyo bw'ubuyobozi
Inzira nyinshi zo kuyobora
Bitangwa binyuze mu gutera inshinge, ntibishobora gutangwa mu kanwa
Umusaruro
Byoroshye gukora mu buryo bw'umusemburo cyangwa mikorobe
Biragoye kandi bihenze mu gukora
Ivumburwa ry'uturemangingo tw'umubiri twa VHH videwo Alpha Lifetech ishobora gutanga uburyo bwo kuvumbura ubudahangarwa bwa VHH
Kuvumbura ubudahangarwa bw'umubiri bwa VHH
VIDEO
SERIVISI YO GUSABA Dore zimwe muri serivisi zijyanye no kuvumbura abarwayi ba VHH. Hitamo ibyo ukeneye: serivisi ikubereye cyane
Ibibazo Bikunze Kubazwa ◢
Q. Antibodies ya VHH ni iki?
A. Antibody ya VHH ni antibody isanzwe ibura urumuri (VL) iboneka mu maraso y'inyamaswa z'ingamiya. Imiterere y'antibody ya VHH ni yoroshye kandi igizwe n'iminyururu ibiri iremereye (VH). Nanobodies zigizwe n'akarere gahinduka k'uruhererekane rw'iminyururu (VHH) gafite uburemere bwa molekile bugera kuri kDa 15. Antibody ya VHH ifite ubushobozi busanzwe bwo kumenya antigens kandi igaragaza ubufatanye bwiza, umwihariko mwiza, ituze ryiza, no kwinjira neza. Antibodys ku miterere yabonetse mu zindi nyamaswa, nka alpacas na sharks. Nanobodies, zirimo agace kamwe gusa gahinduka k'uruhererekane rw'iminyururu iremereye n'uturere tubiri dusanzwe twa CH2 na CH3, nubwo nta gace ka VL kafite, ni antibody imwe ihamye cyane kandi ni igice gito kizwi cyo gufatanya gifite imikorere ya antibody. Alpha Lifetech ishobora guha abakiriya harimo gutegura nanobody, kubaka no gusuzuma isomero rya nanobody, kugaragaza nanobody ya VHH, hamwe na serivisi zitandukanye, binyuze mu buryo bwo gukora nanobody synthesis na phage display, ishobora kubaka no gusuzuma neza harimo ubwoko butandukanye bw'isomero rya antibody ya phage, igaha abakiriya bacu uburyo bwihariye kandi bwo hejuru bw'ibisubizo bya antibody. Dushobora kandi guteza imbere ingamba z’umwuga zo guhindura umubiri w’abakiriya bacu, zishobora guhindurwa nk’umubiri w’abantu, kandi ikigo cyacu gihwanye cyane n’umubiri w’abantu.
Q. Ni ibihe bintu by'ingenzi bigize serivisi yo kuvumbura antibody ya VHH?
A. Ubwa mbere, alpaca idafite indwara iyo ari yo yose, yemerera ubudahangarwa bw’umubiri gukangura ubudahangarwa bw’umubiri wayo kugira ngo bukore ubudahangarwa bw’umubiri, kandi izi nyamaswa zishobora gukora ubudahangarwa bw’umubiri bumwe. Hanyuma, twakuyemo uturemangingo twa B twonyine muri alpaca PBMC, maze RNA yo muri B ikurwamo ihindurwamo cDNA. Amaherezo twakoresheje nk'icyitegererezo maze dusuzuma urukurikirane rwa VHH rwacu hifashishijwe electrophoresis. Hanyuma, twakoze isesengura ry’urukurikirane rw’udutsiko tw ... Amaherezo, binyuze mu ikoranabuhanga ryo kwerekana phage cyangwa ikoranabuhanga ryo kwerekana umusemburo, uturemangingo twa VHH tugaragara ku buso bw’umubiri, hanyuma uturemangingo twa nanoantibodies dushobora gufatanya na antigen twibasiwe tugatoranywa hakoreshejwe uturemangingo twa coated na ELISA. Alpha Lifetech yanakurikiranye kandi igenzura uturemangingo twatoranijwe kugira ngo irebe ko uruhererekane rwa nanobody rukora neza kandi rukora neza.
Q. Akamaro ka serivisi zo kuvumbura antibody za VHH?
A. Ikoranabuhanga rya VHH rikoresha iterambere ritandukanye mu buhanga bw’uturemangingo, kugira ngo gene y’ingenzi isubirwemo kandi ihuzwe hamwe mu bihe bisanzwe, ishobora gutanga igikoresho cyo guhuza cyane n’izindi koranabuhanga, zishobora kuba zirimo tagi zitandukanye, kandi antibody zikorwa n’ikoranabuhanga rya VHH zoroheye gutunganya. Hakoreshejwe uburyo bwo gusuzuma buhanitse, Alpha Lifetech ishobora gusuzuma antibody za VHH zifite imikorere yihariye isabwa n’abakiriya baturutse ku antibody nyinshi mu gihe gito. Hagati aho, antibody za VHH zishobora gukorwa mu buryo butaziguye kandi buhendutse, iyo antibody za VHH zihuye n’ubushyuhe bwinshi n’ibindi bintu bishongesha. Antibody za VHH zishobora gukoreshwa mu buryo bw’uturemangingo kugira ngo zikore izindi mirimo, nko kuba scaffolds, gushyira ibirango, no guhindura aside amine runaka. Antibody za VHH zikwiriye urubuga urwo arirwo rwose rusanzwe rukoresha antibody zisanzwe, nka microtiter plates, electrochemical biosensors, n’ibikoresho byo ku ruhande. Kubera ko antibody za VHH ari ntoya zishobora kugira ubucucike bwinshi mu gice cyo guhuza, ku buryo antibody za VHH zigira inyungu ikomeye mu kongera ikimenyetso bityo zikaba zifite ubushobozi bwo kumva neza. Muri icyo gihe, antibody ya VHH ifite akamaro kanini cyane mu gusuzuma no kuvura ikibyimba, gusuzuma ububyimbirwe, kuvura indwara z’imitsi yo hagati, n’izindi nzego. Antibody za VHH ni ingirakamaro cyane mu kugenzura uburozi bwa mycotoxins mu biribwa no mu biryo kuko byoroshye guhinduranya imiterere y’uturemangingo kandi bifite ubuziranenge buhamye.
Q. Ni gute twahitamo imisemburo y’ubudahangarwa bw’umubiri?
A. Mu gihe cyo gukingira, dukoresha ubwoko butandukanye bw’imyunganire. Dukurikije imiterere y’imyunganire, dushobora kubamo antigen karemano, antigen zisubiramo, antigen zikoze mu buryo bw’ubukorano, na antigen ntoya. Antigen karemano zirimo immunogens za virusi. Uburyo bwo gusukura imyunganire karemano buragoye, kandi bugoye kurusha izindi immunogens, kandi ikiguzi cyo gusukura nacyo kiri hejuru. Imyunganire ya virusi igabanyijemo urukingo rudakora virusi yose, urukingo rw’igice, urukingo rw’ikingira rya virusi, n’urukingo rwa mRNA. Urukingo rwose rudakora virusi rushobora gukangura umubiri kugira ngo ugire ubudahangarwa bw’umubiri, ariko virusi igomba kuba idakora burundu, kandi uburyo bwo kuyitegura buragoye cyane. Ku rukingo rw’igice, twakoresheje poroteyine yo hejuru ya virusi gusa nk’iy’imyunganire, bityo umutekano w’iyi immunogen ni mwinshi, ariko inyongera igomba kongerwamo kugira ngo yongere imbaraga z’ubudahangarwa bw’umubiri. Nyuma yo guhindura imiterere y’uturemangingo, adenovirus na lentivirus bikoreshwa nk’ibyongera imbaraga kugira ngo bitere imbaraga z’ubudahangarwa bw’umubiri binyuze mu kwiyongera no kwigaragaza kwa virusi. Ingaruka z'ubudahangarwa bw'umubiri zirakora neza kandi ziramba, ariko inzira yo kuzitegura iragoye, bityo tugomba kugenzura neza umutekano w'ibinyabuzima. Nubwo mRNA ishobora kwinjizwa mu turemangingo mu buryo butaziguye, kugira ngo uturemangingo tubashe kugaragaza antigen no gushishikariza ubudahangarwa bw'umubiri, iyi nzira iragoye cyane, ibi bikaba bidusaba kwemeza ko mRNA ihamye kandi ikora neza. Nubwo antigens za recombinant zitandukanye mu miterere n'inyito karemano, zishobora gukorwa mu nganda nyinshi. Poroteyine nto cyangwa antigens za polypeptide, zishobora gutegurwa hakoreshejwe uburyo bwo gukora in vitro. Imiterere yazo irashobora kugenzurwa, ariko ishobora kugorana mu miterere. Antigens nto za molecule ahanini ni utuntu duto twa molecule nka peptides na nucleotides, kuko zo ubwazo ntizitera ubudahangarwa bw'umubiri, bityo zishobora gukoreshwa gusa nk'imyuka y'umubiri nyuma yo guhuzwa n'abatwara macromolecular.
Q. Ibyiza by'imiterere ya VHH?
A. Uburemere bwa molekile za nanoantibodies ni buto cyane, akenshi buri hagati ya 12-15 kDa, kimwe cya cumi cy'umusemburo gakondo wa IgG. Imiterere ya kristu ni umupira wa rugby ufite umurambararo wa 2.5 nm n'uburebure bwa 4.2 nm. Imiterere yihariye ya molekile ituma ingirabuzimafatizo zinjira neza, igice cy'ubuzima gito, no kugera ku mpyiko, binyuze mu muyoboro w'amaraso n'ubwonko. Nanoantibodies zigizwe n'uturere twuzuzanya n'utundi duce tw'umugongo. Uduce twuzuzanya harimo CDR 1, CDR 2, na CDR 3. Uburebure bwa aside amine 3 kugeza kuri 28 mu gace ka CDR 3 butuma isomero rya nanoantibody rishobora kubika neza. Ugereranyije n'agace k'umusemburo gakondo ka CDR 3 gafite aside amine 8 kugeza kuri 15 gusa, amwe mu duce twa CDR 3 dufite CDR 3 ndende yadufashije kumenya epitopes zihishe ku buso bwa antigen. Agace k'umugongo k'uturemangingo tw'umubiri karimo FR 1, FR 2, FR 3, na FR 4, hamwe n'impinduka enye za aside amine zikomoka kuri hydrophilic kuri FR 2, kandi iyi mpinduka yongera ubushobozi bwo gushonga kw'uturemangingo mu mazi. Umugozi wihariye wa disulfide hagati ya CDR 1 na CDR 3 wongera ubushobozi bwo kudahungabana kw'uturemangingo mu gihe cy'umuvuduko mwinshi, ubushyuhe bwinshi, denaturant, n'ibindi, ibyo bikaba byorohereza mu gukora no kubungabunga uturemangingo tw'umubiri, kandi binatuma habaho uburyo bushya bwo gutanga.
Niba ufite ikibazo icyo ari cyo cyose, nyamuneka vugaTwandikire igihe icyo ari cyo cyose.
