VHH Single Domain Antibody Discovery Service
Semutengesi ane hunyanzvi mubasa rekugadzira ma "phage display" uye ma "antibody engineering", Alpha Lifetech inogona kupa kugadzirwa nekukurumidza kwema "single domain antibody libraries".
Kumbira Mutengo
VHH Antibody Discovery Service
Alpha Lifetech Inc.inogona kupa kugadzirwa nekukurumidza kwe single-domain antibody (VHH antibody). Matekinoroji edu ekuratidza phage anobvumira masayendisiti kutsanangura zvinodiwa zvavo pakutaura uye kutsanangura hunhu.
Alpha Lifetech Inogona kupa
Kuvaka nekuongorora Raibhurari yeVHH Immune
Masayendisiti ekuAlpha Lifetech Inc. vane ruzivo rwakakura mukuvaka nekubudisa ma antibodies akabatana zvichibva papuratifomu yedu yekuratidza phage. Raibhurari yema antibody e single-domain yakadzivirirwa yakakodzera kugadzirwa kwema antibodies e high-affinity uye antigen-special VHH kana NAR V, nokudaro kudzivirira nguva yakawanda ye antibody affinity maturation effort. Pano, tinogadzira raibhurari yema antibody e single-domain yakadzivirirwa ye antigen-special yevatengi vedu kubva kuma alpacas, ngamera, llamas kana shark dzakadzivirirwa, uye kunyange vanhu (PBL cells activated by antigens in vitro). Ne reverse transcription uye polymerase chain reaction, raibhurari yema antibodies e single-domain ane mamiriyoni 10-100 clones inogadzirwa nguva dzose.
Kuvaka nekuongorora Raibhurari yeVHH yekugadzira zvinhu
Maraibhurari ekudzivirira maantibody emunzvimbo imwe chete anowanzo gadzirwa kubva kuCDR1 neCDR3 yeVHH kana VNAR isina ruzivo. Raibhurari iyi yekudzivirira maantibody inogadzirwa nenzira yekufungidzira iraibhurari isina ruzivo uye ine kuoma kukuru kwe3x10^10 VHHs zvichiita kuti pave nekusiyana kunogutsa. Hazvina ruzivo saka hazvidi kubaiwa llama kana shark. Izvi zvinotibatsira kuchengetedza nguva yakawanda yekuwana antibody chaiyo uye zvinoita kuti pave nekusarudzwa kwemaantibody emapuroteni asina immune kana akachengetedzwa. Maraibhurari akadaro ekugadzira anomiririra sosi yakanaka yemaantibody emunzvimbo imwe chete anorwisa maantigen ega, asiri immunogenic, uye ane chepfu sezvo maraibhurari acho anowanzo kuve akakura zvakakwana uye akasiyana-siyana. Kana kusarudzwa kwemaantibody emunzvimbo imwe chete anonyungudika zvakanyanya kuri dambudziko, takagadzira shuttle phagemid vector system, kusanganisira ma phagemid akati wandei, anogona kuratidza maantibody emunzvimbo imwe chete muE. coli (mbiriso) kana kuaratidza pamusoro pemaparticles ephage.
Zvichakadaro, Alpha Lifetech inopa maraibhurari eScFv Antibody akagadzirira kushandiswa, anoratidzwa nephage kuti akurumidze kuwana maantibody ako. Akagadzirwa kuti agutse zvinodiwa pakutsvagisa kwakasimba, maraibhurari edu anopa Kugadzikana Kwakasarudzika, Kusiyana-siyana Kwakawanda, uye Kugona Kwakakura neKusarura Kushoma. Dzinobva kumarudzi akasiyana-siyana, anosanganisira Munhu, Mbeva, neTsuro, dzinopa hwaro hwakasimba hwekuvandudza kurapa nekuongorora.
| Rudzi | Mhando dzemhuka | Saizi yeRaibhurari |
| Naive scFv | Munhu | 108 kusvika pa1010
|
| Naive scFv | Gonzo | 108 kusvika pa1010
|
| Naive scFv | Tsuro | 108 kusvika pa1010
|
|
|
|
|
|
Sei Uchifanira Kusarudza Isu
Tinogona kupa masevhisi akazara ane chekuita ne nanobody, anosanganisira kubaya mhuka (kusanganisira alpacas, ngamera, nezvimwewo), kuvaka nekuongorora raibhurari yeVHH, kuratidza nekusimbisa ma antibodies eVHH, kuita kuti ma antibodies eVHH ave munhu, nezvimwewo.
Chii chinonzi VHH Single-Domain Antibody?
Antibody yenzvimbo imwe chete (sdAb), kana kuti nanobody, VHH, chikamu cheboka re recombinant antibody fragments, inomiririra antibody diki diki yakaratidzwa kuti inobatsira zvikuru. Nehuremu hwemamorekuru hwe12-15 kDa, ma antibodies enzvimbo imwe chete, ayo anoumbwa chete ne single heavy-chain variable domain uye mainjiniya kubva ku single monomeric variable domains ye camelids' heavy-chain antibody (VHH) kana cartilaginous fish' IgNAR (VNAR) pasina kurasikirwa nekugona kweantigen-binding ability.
Ma antibodies e-single-domain ane zvakanakira zvakawanda: Kubatana kwakanyanya nema antibodies madiki, hunhu hwakanaka hwe physicochemical, uye nyore kushandisa hunyanzvi hwekutanga. Hunhu hwe-single-domain uye nyore kushandisa ma gene e-single-domain antibody zvinoita kuti ive yakakodzera kukura kwe-affinity in vitro.
Kuenzanisa pakati peVHH single domain antibody neconventional antibody
| Mushonga wekudzivirira chirwere weDomain Imwe Chete | Antibody Yakajairika |
Saizi | Duku, nzvimbo imwe chete ine cheni inorema (V)HH)~13 kDa | Cheni hombe, dzinorema uye dzakareruka~120-150 kDa |
Nzvimbo Inosunga Antigen | Monomeric VHChikamu chidiki cheH chinodiwa pakusunga maantigen | Maketani ese ari maviri anodiwa kuti antigen isunge uye igadzikane |
Pasi perwizi | Inonyatsokodzera mainjiniya ekumashure | Kuchinjika kwakaderera kweinjiniya nekuda kwechimiro chakaomarara |
Kugadzikana | Inochengetedza kugadzikana uye simba pa pH yakanyanya uye tembiricha | Haikwanise kutsungirira pH yakanyanyisa kana tembiricha |
Nzira yekutonga | Nzira dzakawanda dzekutarisira | Inopihwa nejekiseni, haigone kupiwa nemuromo |
Kugadzirwa | Zviri nyore kugadzira mune mbiriso kana mabhakitiriya | Zvinonetsa uye zvinodhura kugadzira |
Kuwanikwa kwemasoja ekudzivirira chirwere eVHH vhidhiyo
Alpha Lifetech Inogona Kuwana VHH Antibody Discovery
Kuwanikwa kweVHH Antibody
BASA ROKUKURUDZIRA
Heano mamwe masevhisi ane chekuita neVHH Antibody Discovery. Sarudza zvaunoda: sevhisi yakakunakira iwe
Mibvunzo Inowanzo bvunzwa ◢
-
Mubvunzo. Chii chinonzi VHH antibody?
A. VHH antibody iantibody yechisikigo isina chiedza (VL) inowanikwa muropa remhuka dzengamera. Maumbirwo eantibody eVHH ari nyore uye ane macheni maviri anorema (VH). MaNanobodies ane nzvimbo inorema inorema (VHH) ine huremu hwemamorekuru hunosvika 15 kDa. VHH antibody ine kugona kwekuziva maantigen uye inoratidza hukama hwakanaka, hunhu hwakanaka, kugadzikana kwakanaka, uye kupinda kwakanaka. MaAntibody echimiro ichi akawanikwa mune dzimwe mhuka, dzakadai sealpacas neshark. MaNanobodies, ane nzvimbo imwe chete inorema inorema uye nzvimbo mbiri dzeCH2 neCH3, kunyangwe dzisina nzvimbo yeVL, iantibody yakagadzikana zvikuru uye chikamu chidiki chinozivikanwa chekubatanidza chine antibody activity. Alpha Lifetech inogona kupa vatengi kugadzirira nanobody, kuvaka nekuongorora nanobody library, VHH nanobody expression, uye nhevedzano yemasevhisi, kuburikidza nepuratifomu yekugadzira nanobody synthesis nephage display, inogona kuvaka nekuongorora zvinobudirira kusanganisira mhando dzakasiyana dze phage antibody library, kupa vatengi vedu mhinduro dzakananga, dzakanyanya, dzakasimba dzeantibody. Tinogonawo kugadzira nzira dzehunyanzvi dzekuita kuti ma antibodies ave evanhu kune vatengi vedu, dzinogona kuitwa kuti vanhu vave vanhu, uye kambani yedu inenge yakafanana ne ma antibodies evanhu.
-
Mubvunzo. Ndezvipi zvinhu zvikuru zvinoitwa pakutsvaga maantibody eVHH?
A. Kutanga, alpaca isina chirwere, inobvumira ma immunogens kukurudzira immune system yayo kuti igadzire mhinduro yekudzivirira muviri, uye mhuka idzi dzinogona kugadzira ma single-domain antibodies. Zvadaro, takaparadzanisa ma single B cells kubva ku alpaca PBMC, uye RNA kubva ku ma B cells yakabviswa ndokunyorerwa kuita cDNA. Pakupedzisira takazoshandisa se template uye takaongorora target VHH sequence yedu ne electrophoresis. Tevere, takaita sequencing analysis ye screened VHH antibody sequence, iyo inogona kuvandudza kugadzikana uye hukama hwe prepared VHH antibody. Panguva imwe chete, tinogona zvakare kuita site-directed mutagenesis uye stability screening ye VHH antibodies. Tinogona kuwana higher stability ye VHH antibody mutants nekuisa nzira dzakakosha dzakadai semamwe ma specific mutations ekuongorora. Mainjiniya akabva aisa mhedzisiro ye VHH sequence mu akakodzera expression vector (senge plasmids, ma virus, nezvimwewo) ye VHH antibody expression. Pakupedzisira, kuburikidza netekinoroji yekuratidza mabhakitiriya kana yeast display technology, ma antibodies eVHH anoratidzwa pamusoro pemugadziri, uye ma nanoantibodies anogona kusunga antigen chaiyo anosarudzwa nekushandisa ma coated antigens neELISA. Alpha Lifetech yakarongedzawo uye yakasimbisa ma nanobody akasarudzwa kuti ive nechokwadi chekuti ma nanobody sequences anopiwa vatengi akarurama uye anoshanda.
-
Mubvunzo. Kubatsirwa kwemabasa ekuwana maantibody eVHH?
A. Tekinoroji yeVHH inoshandisa mukana wekufambira mberi kwakasiyana-siyana muinjiniya yemajini, kuitira kuti jini rinotarirwa rigobatanidzwazve uye rigadzirwe pamwe chete mumamiriro ezvinhu akajairwa, izvo zvinogona kupa chishandiso chinosunga zvakanyanya kune mamwe matekinoroji, anogona kunge aine ma tag akasiyana, uye ma antibodies anogadzirwa netekinoroji yeVHH ari nyore kuchenesa. Ichishandisa nzira dzekuongorora dzakanyanya, Alpha Lifetech inogona kuongorora ma antibodies eVHH nemabasa chaiwo anodiwa nevatengi kubva kuhuwandu hwakawanda hwema antibodies munguva pfupi. Panguva iyi, ma antibodies eVHH anogona kugadzirwa nekusingaperi uye nehupfumi, kana ma antibodies eVHH akasangana nekupisa kwakanyanya uye zvimwe zvinonyungudutsa. Ma antibodies eVHH anogona kushandiswa nemajini kuti agadzire mamwe mashandisirwo, akadai sekuva scaffolds, kunyora mazita, uye kushandura maamino acid chaiwo. Ma antibodies eVHH akakodzera chero puratifomu yakajairika ichishandisa ma antibodies echinyakare, akadai sema microtiter plates, electrochemical biosensors, uye lateral flow devices. Nekuda kwehukuru hudiki hwema antibodies eVHH anogona kuva nehuwandu hwakanyanya munzvimbo yekusunga, zvekuti ma antibodies eVHH ane mukana wakakura mukuwedzera kwechiratidzo uye nekudaro kunzwa kwakanyanya. Panguva imwe chete, VHH antibody inoshandiswa zvakanyanya mukuongorora nekurapa bundu, kuongorora kuzvimba, kurapa zvirwere zvepakati petsinga, nedzimwe nzvimbo. VHH antibodies dzinonyanya kubatsira pakutarisa mycotoxins muchikafu nechikafu nekuti dzinogadziriswa zviri nyore majini uye dzine kugadzikana kwakanaka.
-
Mubvunzo. Tinosarudza sei ma immunogens?
A. Mukuita kwekudzivirira, tinoshandisa ma immunogen akasiyana-siyana. Zvichienderana nehunhu hwema immunogens, tinogona kuva ma antigen echisikigo, ma antigen akagadziriswazve, maantigen ekugadzira, uye ma molecule madiki ma antigen. Ma antigen echisikigo anosanganisira ma immunogens ehutachiona. Maitiro ekuchenesa ma immunogens ehutachiona akaomarara, ayo akaomawo kupfuura mamwe ma immunogens, uye mutengo wekuchenesa wakakwirawo. Viral immunogen yakakamurwa kuita whole virus inactivated vaccine, subunit vaccine, viral vector vaccine, uye mRNA vaccine. Mushonga wese we virus-inactivated vaccine unogona kukurudzira muviri kuti ugadzire mhinduro yekudzivirira muviri, asi hutachiona hunofanirwa kuvharwa zvachose, uye maitiro ekugadzirira akaomarara. Kune subunit vaccine, takashandisa virus surface protein chete se antigen, saka kuchengetedzeka kwe immunogen iyi kwakakwira, asi adjuvant inoda kuwedzerwa kuti iwedzere simba rekudzivirira muviri. Mushure mekuchinja kwemajini, adenovirus ne lentivirus zvinoshandiswa sevectors kuti zvikonzere kuitika kwemhinduro yekudzivirira muviri kuburikidza nekudzokororwa kwehutachiona uye kutaura. Kushanda kwekudzivirira muviri kunoshanda uye kunogara kwenguva refu, asi maitiro ekugadzirira akaomarara, saka tinofanira kudzora zvakanyanya kuchengetedzeka kwemuviri. Kunyange zvazvo mRNA ichigona kupinzwa zvakananga mumasero, kuitira kuti masero agone kuratidza antigen uye kukurudzira kuitika kwemhinduro yekudzivirira muviri, maitiro aya akaoma, izvo zvinoda kuti tive nechokwadi chekuti mRNA yakagadzikana uye inoshanda zvakanaka. Kunyangwe maantigen e recombinant akasiyana muchimiro nemaantigen echisikigo, anogona kugadzirwa zvakanyanya mumaindasitiri. Mapuroteni madiki emamorekuru kana maantigen e polypeptide, anogona kugadzirwa nenzira dzekugadzira in vitro. Maumbirwo awo anogona kudzorwa, asi anogona kuoma mukugadzirwa. Maantigen madiki emamorekuru kazhinji mamorekuru madiki akadai se peptides uye nucleotides, nekuti haana immunogenic ega, saka anogona kushandiswa chete se immunogens mushure mekubatana ne macromolecular carriers.
-
Mubvunzo. Mabhenefiti echimiro cheVHH?
A. Huremu hwemamorekuru e nanoantibodies hudiki kwazvo, kazhinji hunosvika 12-15 kDa, chikamu chimwe chete mugumi che antibody yetsika yeIgG. Chimiro chekristaro ibhora rerugby rine dhayamita inosvika 2.5nm uye kureba kweinenge 4.2nm. Chimiro chemorekuru chakasiyana chinobvumira kupinda zvakanaka kwenyama, hafu yehupenyu hupfupi, uye kubviswa kwakanyanya kweitsvo, kuburikidza nechivharo cheropa-uropi. Nanoantibodies ine nzvimbo dzinoenderana uye nzvimbo dzemusana. Nzvimbo dzinoenderana dzinosanganisira CDR 1, CDR 2, uye CDR 3. Hurefu hwemaamino acids 3 kusvika 28 munzvimbo yeCDR 3 hunovimbisa kugona kwekuchengetedza kwe nanoantibody library. Zvichienzaniswa nenzvimbo yeantibody yetsika yeCDR 3 ine maamino acids 8 kusvika 15 chete, mamwe ematunhu eCDR 3 ane CDR 3 yakareba akatibatsira kuona ma epitopes akavanzwa pamusoro peantigen. Dunhu remusana rema nanobodies rinosanganisira FR 1, FR 2, FR 3, uye FR 4, rine hydrophilic amino acid mutations ina paFR 2, uye shanduko iyi inowedzera kunyungudika kwemushonga wekudzivirira muviri mumvura. Chisungo chakakosha che disulfide pakati peCDR 1 neCDR 3 chinowedzera kugadzikana kwema antibodies pasi pekumanikidzwa kwakanyanya, tembiricha yakakwira, denaturant, nezvimwe zvinhu, izvo zvinobatsira kugadzirwa nekuchengetedzwa kwema nanobodies, uye zvinogadzirawo mukana wekushandisa nzira itsva dzekupa.
Kana muine mibvunzo, ndapota inzwai makasununguka kupindurataura nesuchero nguva.
