Nhanganyaya Yebasa reKuratidza Mapeji

Kuratidzwa kwePhage, senzira yemamorekuru yakavakirwa pakuchinja kwemajini ephage DNA, kwave chishandiso chine simba remamorekuru chekugadzira mamorekuru ekuongorora zvinhu zvakanangana nezvinangwa zvakananga nekuisa zvidimbu izvi pamusoro pephage uye nokudaro kusarudza zvidimbu zvepeptide/antibody zvine hunhu hwakasiyana hwekusunga kubva mumhando dzakasiyana-siyana (maraibhurari).

Alpha Lifetech ine makore gumi eruzivo mukugadzira ma antibodies e phage display uye inogona kupa basa rehunyanzvi rekuvaka uye kuongorora ma antibodies library kune vatengi vedu. Zvichienderana nezvinodiwa nevatengi, tinogona kupa mhando dzakasiyana dzema antibodies library, dzakadai sema immune libraries, ma native libraries, ma synthetic libraries uye semi-synthetic libraries. Tekinoroji ye Phage display library, iyo inobatanidza ma antibodies genes nema gene ema phage coat proteins akadai seM13, ndiyo tekinoroji inonyanya kushandiswa pakugadzira ma antibodies e recombinant uye inoshandiswa zvakanyanya mukugadzira ma antibodies chaiwo.

Alpha Lifetech ichabaya majekiseni ealpaca, llama, alpaca, neshark, nezvimwewo, kuitira kuti kushanda kwawo kusvike 10^5. Tichatumira mushumo wekuyedza kushanda zvakanaka kumutengi kuti tive nechokwadi chekuti data racho rakarurama uye nderechokwadi. Maraibhurari emuno ane mabhenefiti ekuti ane chepfu shoma uye kuti haana simba rekudzivirira muviri. Alpha Lifetech ine maraibhurari akawanda emuno akagadzirwa kare, anogona kushandiswa zvakananga pakuongorora majekiseni emuviri pasina kubaya mhuka, nokudaro zvichipfupisa nguva yepurojekiti, zvichifambira mberi kufambira mberi kwekuyedza kwemutengi, uye zvichivimbisa kushanda zvakanaka kwejekiseni remuviri.

kuwanikwa kwemasoja ekudzivirira chirwere-Alpha Lifetech

Mufananidzo 1: Pfungwa yekuratidza ma "phage"

Sisitimu Yedu Yekuratidza Maphage

Ruzivo rukuru rweM13/T4/T7 phage display system rwunoratidzwa muTafura 1.

Tafura 1 Mota dzakasiyana dzekuratidza ma phage uye hunhu hwadzo
	M13	T4	T7
Saizi yeGenome	6407 bp	168895 bp	39937 bp
Ratidza Mapuroteni	pVI, pIII uye pVIII	SOC neHOC	gp10B
Saizi Yekuratidzira	>110 kDa is pIII
Saizi Yekuratidzira
Kuwanda kweZviratidzo		Makopi anopfuura 810 paSOC
Kuwanda kweZviratidzo	Makopi anopfuura mazana maviri nemazana manomwe pa pVIII	Makopi anopfuura 155 paHOC
Kutenderera Kwehupenyu	Lysogeny	Lytic	Lytic

Alpha Lfetech Inogona Kupa

Kuratidzwa kwePhage kunobatanidza genotype ne phenotype kuita chimwe chete, zvichibatanidza kusarudza nekuwedzera simba nehunyanzvi hwekuongorora. Alpha Lifetech inogona kupa mabasa akasiyana-siyana ekugadzira maantibody ephage display.
Mabasa makuru anosanganisira: chikuva cheraibhurari yeVHH antibody, chikuva cheraibhurari yeScFv antibody, chikuva cheraibhurari yeFab antibody, chikuva chekuvaka raibhurari yephage, chikuva chekuongorora raibhurari yephage uye sevhisi yehumanization yeantibody nemamwe mabasa.

kugadzirwa kwemasoja ekudzivirira chirwere - Alpha Lifetech

Mufananidzo 2: Maitiro ekugadzira chiratidziro chePhage

Maitiro Ekugadzira Antibody yePhage Display

Tinogona kubaya mhuka mapuroteni anowanikwa murabhoritari yedu kana anopiwa nevatengi vedu, kusanganisira mapuroteni akasiyana-siyana e macromolecular, peptides uye membrane proteins. Kuburikidza nekuyedza kushanda kweELISA, vaongorori vedu vakaita PBMC isolation uye RNA extraction kubva muropa realpacas nemhedzisiro yakanaka yekudzivirira. Nekudzvanya reverse transcription uye gene amplification, takagadzira gene iri pa pMESC, pComb3XSS kana pCANTAB 5E phage vectors. Nekushandura masero ane TG1 E. coli electrotransformation, maraibhurari ephage antibody ane simba reraibhurari rinodarika 10^9 anogona kuwanikwa. Maraibhurari eantibody ane simba guru reraibhurari ane kusiyana kwakanaka uye anobatsira kuwana ma antibodies ane hukama. Kuti tive nechokwadi chekuti raibhurari yeantibody ine mhando, tichatsanangura kugona kweraibhurari uye mwero wekuisa raibhurari. Pakupedzisira tinoronga maraibhurari kuti tipe vatengi vedu ma sequences anodiwa 30-50. Zvadaro, tinoita kuongorora raibhurari yekuratidza phage, Kutanga, antigen inomiswa kufamba pama polypropylene microplates, uye kuburikidza nekuongorora katatu kusvika kashanu, ma phage-antibodies ane simba rekusunga asina simba anobviswa, uye ma clones chaiwo anobatana ne antigen anochengetwa. Pakuratidza phage, nzira yeELISA inoshandiswa kuguma ne clone yakanaka, nepo pakuratidza yeast, nzira yeFACS inoshandiswa kuita kuti masero aratidze zvidimbu zve recombinant antibody tichishandisa ma antigen ane labeled. Kuburikidza nekuongorora, tinogadzira ma antibodies ane high sensitivity, specificity uye high affinity.

Mufananidzo 3 Maitiro ekugadzira maantibody ekuratidza phage

ZVAKANAKA ZVEBASA
Zvakanakira zvebasa

Alpha Lifetech yakazvipira zvikuru kune tekinoroji yekuratidza ma phage, uye inogona kupa vatengi masevhisi akawanda uye ari nani e phage.

Nguva uye kushanda zvakanaka kwazvo

Kambani yedu inogona kutsvaga ma antibodies anogutsa munguva pfupi; tinogona kuita bvunzo dzakasiyana-siyana dzekuongorora ma antibodies zvichienderana nezvinodiwa nevatengi.

Hunhu hwepamusoro hwechigadzirwa

Kambani yedu inobatsira vatengi kugadzira masoja ekudzivirira chirwere ane hungwaru hwakanyanya uye kugadzikana, zvichiwedzera kubuda kwemhedzisiro yekutsvagisa kwesainzi.

Ipa sarudzo dzakasiyana-siyana

Kambani yedu inogona kuvaka maraibhurari emasoja emhando dzakasiyana-siyana (vanhu, mbeva, tsuro, alpaca, nezvimwewo) uye inogona kuvaka mhando dzakasiyana-siyana dzemaraibhurari emasoja ekudzivirira chirwere (scfv, Fab, VHH, nezvimwewo).

Kukwanisa Kukuru

Raibhurari yedu yema antibodies ine simba guru rema antibodies anopfuura 10^9

Mibvunzo Inowanzo bvunzwa-Matambudziko akajairika ekuratidza phage

Mibvunzo yekuratidza phage ◢

Mubvunzo.
Chii chinonzi tekinoroji yekuratidza phage?

A.
Tekinoroji yekuratidza phage ndeyekuisa gene rekune imwe nyika munzvimbo chaiyo yeproteyini yephage capsid, uye kuratidza gene rekune imwe nyika pamusoro pephage pasina kukanganisa mashandiro akajairika ephage. Mushure mekuongorora kakawanda, protein yekune imwe nyika ine hunhu hwakasimba uye inoshanda zvakanyanya muhupenyu yakawanikwa pakupedzisira.
Mubvunzo.
Ndezvipi zvakanakira tekinoroji yekuratidza phage?

A.
Maraibhurari emasoja ane simba guru anogona kugadzirwa uchishandisa tekinoroji iyi. Kuongororwa kwenguva pfupi, mashandiro acho ari nyore, uye huwandu hwemashandisirwo acho hwakakura, zvichibvumira kuzivikanwa kwematanho akakodzera emapeptide kana mapuroteni akatarwa. Mumashandisirwo ane chekuita nehutachiona, vaongorori vanogona kuwana masoja ekudzivirira chirwere anobatana nema epitopes akakosha ehutachiona nekuongorora maraibhurari makuru emasoja ane simba. Maitiro aya anogona kukanganisa kupinda, kudzokororwa, kana nzira dzekudzivirira hutachiona, nokudaro zvichigadzira hwaro hwekurapa chirwere.
Mubvunzo.
Ndeupi musiyano uripo pakati pe hybridoma technology ne phage technology?

A.
Tekinoroji yekuratidza mabhakitiriya inoshandiswa zvakanyanya kupfuura tekinoroji yehybridoma, iyo inongogumira pakugadzira huwandu hushoma hwemasoja ekudzivirira chirwere anorwisa ma immunogens chaiwo. Kusiyana neizvi, tekinoroji yekuratidza mabhakitiriya inogona kupa raibhurari yese yemasoja ekudzivirira chirwere anobva kumarudzi akasiyana-siyana emasoja ekudzivirira chirwere.
Mubvunzo.
Ndeipi nzira yekushandisa tekinoroji yekuratidza phage?

A.
Tekinoroji yekuratidza ma "phage display" ine mashandisirwo akasiyana-siyana, anosanganisira kuongorora ma "receptors" e "pathogenic microorganisms", kudzidza kudyidzana kwemapuroteni, kutevera kutapurirana kwemasaini pakati pemasero, kuongorora zvirwere zvemhuka, uye kugadzira majekiseni, pakati pezvimwe zvinhu.
Mubvunzo.
Tarisiro yetekinoroji yekuratidza phage?

A.
Tekinoroji yekuratidza ma "phage display" ine mabhenefiti akawanda uye inogona kushandiswa munzvimbo dzakasiyana siyana. Zvisinei, dambudziko guru mukugadzira ma "recombinant antibodies" uchishandisa nzira yekuratidza ma "phage display" ndiko kuoma uye mutengo unoenderana nemaitiro acho. Maitiro aya anoda kubatanidzwa kwemaitiro e "molecular biology", anosanganisira mutation, cloning, uye expression, pamwe nekuongorora kwe "immunological, biochemical, uye biophysical" kuti zvionekwe uye zvionekwe ma "antibodies" anodiwa.

Mibvunzo ine chekuita neAntigen ◢

Mubvunzo.
Ndedzipi nzira dzekuratidzwa kweantigen?

A.
Kuratidzwa kweAntigen chinhu chakakosha zvikuru. Nzira dzekuratidzwa kweantigen dzinosanganisira kuratidzwa kweantigen zvakananga, kuratidzwa kweantigen zvakananga, kuratidzwa kweantigen kuburikidza nekuongorora masero ese, kuratidzwa kweantigen kuburikidza neliposomes, nano-phospholipid disks uye VLPs.
Mubvunzo.
Ndeapi musiyano uripo pakati pemhando dzakasiyana dzemaantigen?

A.
Nzira yekuputira zvakananga iri nyore, asi chimiro cheantigen chiri nyore kuchinja, uye mashandiro ekuputira zvisina kunanga akaoma, asi zvinogona kuchengetedza chimiro cheantigen uye kuvandudza kushanda kwekuratidzira. Kuongorora masero kunogona kutevedzera mamiriro chaiwo ari mumuviri, asi pane dambudziko rekusungana kusingatsananguriki. Nano-phospholipid disc ine saizi diki, kushanda kwepamusoro uye kugadzikana kwakanaka; VLPs inosanganiswa neantigen kana kuputirwa neantigen, ine immune genicity yakakura uye kuchengetedzeka kwakanaka, uye inogona kukonzera mhinduro yakasimba yekudzivirira muviri.
Mubvunzo.
Chii chinoita kuti huwandu hweantigen huvepo pakuongorora mhedzisiro ye phage display technology?

A.
Kana huwandu hwacho hwakanyanya kuderera, mukana wekuti munhu abatane ne phage uchaderera, zvichikonzera kudzikira kwekushanda kwekuongorora. Kusiyana neizvi, huwandu hwacho hwakanyanya hunogona kuwedzera mukana wekuti munhu abatane zvisina tsarukano, kukwidza chiratidzo chekumashure, uye kukanganisa kuzivikanwa kwema clones akanaka.
Mubvunzo.
Tingava sei nechokwadi chekuti antigen inoramba iine chimiro chayo chemuzvarirwo panguva yekuongorora?

A.
Nzira yekugadzirisa zvinyoro inogona kushandiswa kudzivirira kupisa kwakanyanya, acid yakasimba uye alkali. Nzira yekuvhara isina kunanga uye biotin-streptavidin system zvakashandiswa kuderedza kukuvara kwechimiro cheantigen. Kana antigen iri protein yemembrane, inogona kuratidzwa pamusoro pemasero asina kubatwa kana kushandiswa kutevedzera nharaunda yemembrane, senge nano-disk, kuchengetedza chimiro chayo chechisikigo chemembrane protein.
Mubvunzo.
Nzira yekusimbisa sei kubatana kwema antibodies akaongororwa ne phage display technology kuma antigens?

A.
Pamusoro pekuedza kwakajairika kwakadai seELISA, Western Blotting (WB), uye Immunofluorescence (IF), matekiniki ekuona Surface Plasmon Resonance (SPR) uye Bio-Layer Interferometry (BLI) anogona kushandiswa kutarisa maitiro ekubatanidza nekuparadzanisa ma antibodies nema antigens munguva chaiyo, zvichibvumira kusarudzwa kweaffinity uye kinetic parameters.

Mibvunzo Inoenderana Nezvirwere zveMuviri ◢

Mubvunzo.
Ndeapi maitiro ekudzivirira muviri emhuka muhunyanzvi hwekuratidza ma phage?

A.
Kuratidzwa kwePhage kunogona kushandiswa kugadzira ma antibodies emhando dzakasiyana-siyana dzinogadzira ma antibodies, kusanganisira asi kwete chete kuvanhu, mbeva, makonzo, tsuro, huku, ngamera, alpacas, mombe, imbwa, makwai, tsoko, uye shark.
Mubvunzo.
Ndezvipi zvinhu uye mamiriro ezvinhu anoshanda emaraibhurari emasoja ekudzivirira chirwere anobva kumarudzi akasiyana emhuka muhunyanzvi hwekuratidza mapoinzi ephage?

A.
Tekinoroji yekuvaka maraibhurari ekudzivirira mabhakitiriya embeva yakasimbiswa zvakanaka, inodhura zvishoma, uye ine nguva pfupi yekuyedza, zvichiita kuti ive yakakodzera kuongororwa kwekutanga kwemasoja emuviri uye tsvakiridzo yekutanga. Masoja ekudzivirira mabhakitiriya etsuro anoratidza hukama hwakasimba uye hune hunyanzvi, ane huwandu hwakakura hwekuziva epitope kana tichienzanisa nema antibodies embeva. Masoja ekudzivirira mabhakitiriya evanhu anogona kuwanikwa zvakananga kubva kumaraibhurari ekudzivirira mabhakitiriya evanhu, nokudaro kudzivirira mhedzisiro inogona kuitika kana masoja ekudzivirira mabhakitiriya akasiyana-siyana akapihwa mumuviri wemunhu. Maitiro aya akakosha mukutsvagisa nekugadzira mishonga, kunyanya mukugadzira masoja ekudzivirira mabhakitiriya ekurapa.
Mubvunzo.
Chii chinonzi general animal immunogen chinoshandiswa mu phage display technology?

A.
Ma immunogen emhuka anowanzo kamurwa kuita mapuroteni antigen, polypeptide antigen, nucleic acid antigen uye pathogen-related antigen, pakati pawo panosanganisira pathogen-related antigen yehutachiona (complete virus particle, virus protein, virus non-structural protein, nezvimwewo), bacterial antigen (complete bacterial extract, cell wall component, secreted protein, toxin, nezvimwewo) uye parasitic antigen.
Mubvunzo.
Tingagadzirisa sei nyaya yekusagona kudzivirira zvirwere mumuviri mumhuka panguva yekudzivirira muviri?

A.
Gadzirisa ma immunogens nekutsiva ma immune antigens nema recombinant proteins uye kutsiva zvidimbu zve fire-antivirus nema structure proteins ema virus. Gadzirisa ekuyedza magadzirirwo uye gadzirisa dose ye immune immune.
Mubvunzo.
Kugona kwekudzivirira muviri kwe immunogens kunogona sei kuvandudzwa mu tekinoroji yekuratidza phage?

A.
Kugona kwekudzivirira muviri kunogona kuwedzerwa nekubatanidza maantigen nemapuroteni anotakura, akadai se bovine serum albumin. Pamusoro pezvo, ma immune adjuvants, anosanganisira Freund's adjuvant nealuminium adjuvant, anogona kushandiswa kuwedzera nguva yekugara mumuviri uye kukurudzira kushanda uye kuwanda kwemasero ekudzivirira muviri, nokudaro zvichivandudza kugona kwekudzivirira muviri.

Mibvunzo yekuvaka raibhurari ◢

Mubvunzo.
Chii chinonzi raibhurari ye phage antibody?

A.
Raibhurari ye phage antibody muunganidzwa wema antibodies akasiyana-siyana e phage akagadzirwa nekuunganidza seti yakazara yemajini enzvimbo dzakasiyana kubva kuma antibodies kuenda kuma phage vectors uye kuaburitsa pamusoro pema phages. Maitiro aya anoguma nekuumbwa kweraibhurari ye phage antibody.
Mubvunzo.
Maitiro ekuvaka raibhurari ye phage antibody

A.
Maitiro ese ekuratidza phage anosanganisira matanho akati wandei akakosha: kubvisa masero eropa eropa (PBMC) kubva kumhuka dzakabayiwa nhomba, kuparadzanisa RNA yese nekuishandura kuita complementary DNA (cDNA), kuwedzera gene iri kutariswa uchishandisa tekinoroji yepolymerase chain reaction (PCR), kuumba gene iri kutariswa kuita phage vector, uye kushandura vector kuita sero remubatsiri kuti rionekwe. Maitiro aya anoguma nekuvakwa kweraibhurari yekuratidza phage, iyo inogona kuona nekukurumidza ma antibodies ane hukama hwakakura. Zvadaro, raibhurari iyi inogona kushanda senzvimbo inokosha yekuvandudza majekiseni, mishonga yekurapa, uye ma reagents ekuongorora.
Mubvunzo.
Ndezvipi zvinhu zvakakosha pakuvaka raibhurari yepamusoro-soro?

A.

Iva nechokwadi chekuti huwandu hwese hwePBMC hwakakwana. Iva nechokwadi chekuti RNA yakanaka pasina kuora.

Kuti zvive nechokwadi chekuti huwandu hwese hwevanoshandura zvinhu hunowanikwa, huwandu hweraibhurari hunofanirwa kunge hwakapetwa kagumi kusvika zana kupfuura kusiyana kwedzidziso.
Mubvunzo.
Ndezvipi zvikamu zveraibhurari ye phage antibody?

A.
Zvichienderana nekwakabva raibhurari yeantibody, maraibhurari ephage antibody anogona kuiswa muzvikamu zviviri: maraibhurari e immune phage antibody uye maraibhurari e native phage antibody. Musiyano mukuru pakati pemhando mbiri idzi ndewekuti mhuka dzakabayiwa here. Pamusoro pezvo, maraibhurari e native antibody anogona kugadzirwa kuburikidza ne semi-synthesis kana synthesis.
Mubvunzo.
Munhu angaongorora sei kunaka kweraibhurari ye phage antibody?

A.
Kune zviratidzo zviviri zvikuru zvekuongorora mhando yeraibhurari yemasoja ekudzivirira chirwere: kugona kuchengetedza uye kusiyana-siyana. Kugona kuchengetedza kukuru, pamwe chete nekusiyana-siyana, kunoita kuti pave nekuongororwa kwakanaka kwemasoja ekudzivirira chirwere ane hukama hwakakura uye hune hunyanzvi.

Mibvunzo yekuongorora raibhurari ◢

Mubvunzo.
Chii chinonzi maitiro ekuongorora raibhurari ye phage antibody?

A.
Mukufamba kwekuongorora kwemuviri, raibhurari yekuratidzwa inoiswa zvigaro zvakatarwa uye yozogezwa zvakanyanya kubvisa ma non-reactive phages. Ma conjugates anowanzo bviswa achishandisa acidic kana high-salt solution uye ozowedzerwa mumasero akakodzera ekuchengetedza kuti awedzere kunaka. Mushure mekuongorora katatu kusvika kashanu, ma antibodies ane high affinity anowanikwa.
Mubvunzo.
Ndedzipi nzira dzinoshandiswa pakuongorora maraibhurari ekudzivirira chirwere chephage?

A.
Nzira dzekuongorora ma "phage" dzinosanganisira kunyanya "solid-phase panning", "liquid-phase screening", uye "cell-based screening". Kusarudzwa kwenzira yakakodzera yekuongorora kunofanirwa kutarisa hunhu hwemamorekuru anotarisirwa, zvinangwa zvekuongorora, uye nzvimbo yerabhoritari.
Mubvunzo.
Ndezvipi zviratidzo zvakanaka nezvakaipa zve phages?

A.
Kuongorora kwakarurama ndiyo nzira inonyanya kukosha yekuongorora ma antibodies e phage anosunga kumapuroteni anotarisirwa. Kuongorora kwakaipa kunobatsira kubvisa ma antibodies asina chaiwo. Munguva yekuongorora ma antibodies e phage, ma antibodies asina chaiwo anogona kuonekwa nekuda kwekukanganiswa kubva kune target antigen label, zvinhu, kana mamwe ma antigen akafanana nemaumbirwo e antigen anotarisirwa. Muzviitiko zvakadaro, kuongorora kwakaipa kunogona kuwedzera kushanda zvakanaka kwekuongorora.
Mubvunzo.
Nzira yekudzivirira sei kuongorora ma antibodies asina kutsanangurwa?

A.

Maantigens akanyorwa nemarudzi akasiyana ezviratidzo anogona kukosheswa pakuongororwa kwekutanga, kana kuongororwa kusina kunaka kunogona kushandiswa kuderedza huwandu hwemaantibody asina kutsanangurwa.

Unogona kusarudza kutsiva mvura yekuvharira yakakodzera kana kushandisa mvura yekuvharira yakasiyana kudzivirira izvi. Kana antibody inoona mutsetse wesero rekumashure (CHO kana HEK293) ikaongororwa, unogona kutsiva mutsetse wesero rekumashure wakasiyana nezvinangwa zvekuongorora kana kuongorora zvisina kunaka.
Mubvunzo.
Munhu angaongorora sei ma antibodies akawanikwa?

A.
Nzira dzekuongorora ma antibodies dzinowanzo sanganisira ELISA, Western Blotting (WB), flow cytometry, immunoprecipitation (IP), immunofluorescence (IF), immunohistochemistry, nedzimwe nzira dzakasiyana-siyana dzekuedza. Alpha Lifetech ine ruzivo rwakakura mukuona ma antibodies uye ine timu yehunyanzvi yekuedza, ichiona kushanda kwema antibodies mukuyedza kunotevera.

Kana muine mibvunzo, ndapota inzwa wakasununguka kutibata chero nguva.

Leave Your Message

Basa Rinonyanya Kuonekwa

Puratifomu Yekuwana MaAntibody