HUE030285T2 - Emberi variábilis régiót tartalmazó immunglobulin hibrid könnyûláncot exprimáló egerek - Google Patents
Emberi variábilis régiót tartalmazó immunglobulin hibrid könnyûláncot exprimáló egerek Download PDFInfo
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- HUE030285T2 HUE030285T2 HUE12195716A HUE12195716A HUE030285T2 HU E030285 T2 HUE030285 T2 HU E030285T2 HU E12195716 A HUE12195716 A HU E12195716A HU E12195716 A HUE12195716 A HU E12195716A HU E030285 T2 HUE030285 T2 HU E030285T2
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Claims (13)
- SZABADALMI IGÉNYPONTOK 1 Egér, amelyben megtalálható: gsnszsgmens{VN), (Da): és k^^f^^iips§h);is^ô' égy vagy több m$m V«, Da è$ Jh génszegmenssel egy endegit egei nehéztánc immuaoglobafi iékuszban; és öt: egy endogén A (iamöcia) kennyőinc étiéi, amely MÍKiy üjra^üteßen emberi lambda vaaiblt(hVA) és egy girarefidezetien emberi iarnbcia kapcsoló (HJA) génszegmenst, ghóí rn egérben megtalálható az emíllelt endogén egér A könnydláne- alléi:: első VA^A-Ck: génkiaszterének deféciőia és az émiíiétt emberi W és JA gemszegmeriseket tartalmazó altét második VA-JA-DA: géhklsszterében lévő VésJA génszegmébsébek -részben, νιί8§^^Μ>©ίΐ:^η#*:ί^^Κ^ί, arnetyek mötéé útón kapesoitók ez említeti második VA-JA-GAgénkiaszter egy érintetlen égét lambda konstans (ÇA) téglához, így ez egér egy :hVA; hJA és egy egér CA génbe! származtatottkönnyéinçæit éstgrlmák bitéi az Enh 2.4, agár lambda .3’ ehhsrseét#rib}::és inti 3.1 éribbgén enharisarak érintettének maradnak az említett A kénrsyûiànc alléidén,
- 2, Az egér m 1 -es igénypontiságlgi, 2h$ t2:$Rds##|imbds).kbnn^ti^'i{i#l fertelme
- 3. Az egér az eddigi igénypontok bármelyike szerint, ahol: ez endogén λ (lambda) kénnyőíánc alléi rendelkezik; a) legalább 12 újrarende2ötlee:.#^di'VA' géftszegrmpiSöi vagy
- 4, Azégéra^ss igénypont szerint, aMaz emberi Sik0m0mm^Méí^mÉ m hVA3-1, «4-3, hv'A2-8, hVA3k| «3-10, wmm bVA3-12 szegmensek; vagy ρμ ηΜΑΖ-ΙΙ, ΑνΑδ-Ιδ. hVAA-18, hVA3-1i, hVA3-2í, hÄ22, ÍWA2-23;: í»2S és hVA3-2? szegmensek,
- 5, Az egér az i-es igémypn? szerint, ehet az endcprr A (lambda) ttónnpíánc alléi egy óprendezetlen JA génszegmenst tartalmaz, amely a hJA1.
- 6. Az egér az 1-es igénypont szeriní, ahol az endogén A fambda) korrnyílíáns alléi: négy djfarendezetlen emberi JA génszeg mensLMaimaz::
- 7, Az iggér a dtéSigényph? szérinbahoi a négy üjmrehdezetisn emberi JA génszegmens aJAi) $%.;<$&M JA7.
- 8. Az egér a? 1-es igénypont szerint, ahoi p endogén Agembda) Kónnyiáoe aílélbői hiányzik egy endogén VA génszégmens.
- 9. Az egérp 1-3, az δ-ës #S*s®;^ft^k}n^ölbfté|Étsziinni afe&é&emfcgii» κ I Q, Az egér az: 1ms igénypont szerint,: ahol az endogén egér A könnyöláne alléi: iartaimez:: a) : agy az emberi A kbrinyölánc lókusztóiyamatös székyennlljéí, amely VAS^Udo! a VA34-ig iedéít. b) sgy az emberi A kénnyöiánö tókusztóiyamatos szekyeneipt, amely YA3-294Ö! a vA3-1-ig terjed: vagy c) egy ipsmbeh λ klrmyiánc folyamatos s^@Kveríeia|st, amely VA3>3Spí :hVA3-:^i|l?pcí és az emberi A kÖpyCtíáne lökusziipiypÄS szekvenciáját, amely vA542~íöí hVAMOdgprjed: ill izeiéit sejt, gme% egy IWÁ, hJh és egy Oh égér génből: szárnazlatmMn^émáeHmmá, ahöká sajt az addig: igénypontok bármelyike szerint meghatározott egérből szárasaik vagy akból Kir^prl»i.,i§#^i:^:ia^§ikeÄ a) égy vagy több nehézlánc variábilis génszegmens (Vh); divarzitás génszegmens (1¾} és kapcsoló gődpsgrnens;(já} sserèje égy vagy több emberi im, Dh és iÍH:0^^^sséf^Sif^rÉ^iÉn:^|éf'-n^éz!|rtó imnwnpgíobpíín fókuszban; és b) agy endogén A {iambds} kPnyíiiánc adiil, amsiy íériimag agy öjrarendezeti emberi lambda variábilis (hVA) és agy újrarendezett emberi gáns^ptábstj, érag ^{teU,eai^|n egér A könnyöiánc alléi alsó Vh-d^úk génklasxterérrek cteléeiéjávai és az emlősé emberi VA és JA génszegmensekét tartalmazó altéi második VA-JA-GA génkíaszterlbea tévő 1-1 génszegmensének - részben, vif ÿ 'égéézbes, llíS^ő .* ih#yéÍ8iÍiéál. áméiysk műtéti «ton kapcsoltak axsafiett mPoőÉ pyA-CA pftkiaszte: egy érititetísri egér CA konstans régióhoz, és ahoi az Inh 2;4vegèr lambda 3' enhancer (Enh) és |nh 3.1 endogén enhancerek érintetlenek maradnak az említett A könnyülánc aiiélben.
- 12. Az izolált sajt a 11-es igénypont szerint, shot a agit egy lihifeoife
- 13. Izolált egér embrionális őssejt (ES), amely tartalmaz: a) egy vagy több nehízino variábilis génszegmens {W}, dlverzitás génszegmens (Dm és kapcsoló génszegmens (J»<j cseréje agy vagy több amben V·,·. Dh és J« géhszégmertssel egy endogén egér oehézlánc Immunoglobulin fókuszban; és b) egy endogén A (lambda) kónnyüiánc aliét, amely tartalmaz agy öjrsmndezetíén emberi lambda variábilis (riVA) és egy ójrarandezetfen emberi lambda kapcsoiő (hJA) génszegmensi, az enimh endogén egér A kőnnyöánc áíiél eisö VA^A-CA génkieszíéréPk delésiója és az említett emberi VA és 3A génszogmensekét tariaimaző alii második VA^A-iA génidasztéréban lévő VA*3A:|lnszegménsének -részben, vagy egészben: történő - pipjéstfését. amelyek mated ötön kapcsoltak ázi; említett második VA^feiSA génklaszter egy érintetten egét CA Iimnstens régióhoz, ahól az Enh 24, egér lambda 3' enhancer: (loh) és Enh 3.1 endogén onhancerek érintődének mátíkpak az emiiiett A könnyöiánc aiiélben. 14 Egér embrió, amely a 13-as Igénypont szerint meghatározott őssejtet tartalmazza, vágy abból fejlődöd ki, vagy abbéi kinyerhető, ahol az egér embrió rendelkezik; iá) egy vagy több: dábéziánc variábilis génszegmens (Ve), ölverziiás génszégmenbliM ésikápcsaiő génszegmena (|fé osoréje egy vagy több emberi Vh, De és p génszegmenssel egy endogén egér oehézlánc immunoglobulin fókuszban; és Ü M Ja#da) könnyöiánc alléi, amely tartalmaz egy újrarendezeden embed: iiambda variábilis (hVA) és égy öjtarepezeden etliberi lambda kapcsoló (hJA) génszegmenst; az említett endogén egér A könnyöiánc alléi első VA-IA-CA géhklasztefénék: déiéelőfS és az emiitett emberi VA és JAfériszegmensekel íartáSmazó eiiálimPodik VA-őÁ^A génkiaskteíébén tévő VA-J.A génszegmensének - részben, vagy egészben történő » helyettesítését, amelyek műtéti ötön kapcsoltak az amiítei második V.WA'CA génklaszter agy érintésén egér CA konstans régióhoz, ahol az Enb 2,4. egér lambda 31 phspoer prih) és Enh 3,1 endogéhiénhencerék érintetiefiek maradnak az. emiitett A könnyülánc aiiélben:, 15, Ä 12-es igénypont s2Ât;rn^bi^^iMÉiÂ':^yân hibridóma sejt létrehozásához, émeiy egy hVA, h JA és égy égér hibridémsrseft· az 1-10 igénypontok bármelyiké szerint meghétárözód agárból származik vagy abból kinyerhető, ahoi a sejt rendelkezik: dlverzitás génszegmens (©η) ős képcsőié génszegmens (P) cseréje egy vagy több emberi nehézláncjmmiínoglóbüíln ishyszöari:; és öjí sgy endogén k (¾¾¾¾ k$ppáfte aliéiiéi, amely tartalmaz; egy újrareadezsttémbeíi iámfesia vwiáfeiíis ss @sgy üjmmnddzed embéíi iiiig kapcsoló (hJA) génszégménsl és az említett endqgén egér A kőnnyőlánö áiiél tâ$-VA<JMP& génkíaszíeréoek cielèeiôjs és az émíiteít emben VÀ és JA génszegmenseket tertelmazc alléi második VÂ-JA-CA génkiasxterében lévő: V>WA génszegmensének - részben. vagy egészben történő - helyettesítésé, amelyek bitéi ötön kapösoíiak az amidéit mäsöikÄJAßAgenklasztsr egér CA konstans régiőboz, és alsói m léh :2,4 «fk lambda t enbanoer poh) és EOhM endogén an· isnoergk énntetiénsK maradnak az említett A kënnÿdiéne ailéSbp,
- 16. Hibridóma sejt. amely egy hVA, hJA és egy CA agár génből származtatott könnyűíáncot exprimât, ahol a hibridóma sejt ax 1 -lö ípnypöftiok bármsípo szerint meghgározott egérböí származik vágy abbot kinyerhető, ahoi a seji i^delezik: aj agy vágy több negézllnc vsÄilis géaszegmens (Vm), öív^á§í^^^^?;|||r) f s-fcpcpé :g#np§gpégő (Jh) cseréj egy vagy több emberi \% Dh és űngéns^gmenssd e§y «näog^r egér nehézlánc Immunoglobulin fókuszban; és b) egy endogén A (lambda) köhnyűíáns siléiloi, amely tartalmaz égy újrarendezett emberi lambda variábilis (hVA; és agy újrarendezett embed lambda kapesolô (M}. iénszegmaost; és az említett endogén egér A könnyűién« alléi első VA-JACA genkfasztsífnek deléctója és az említett lévő νΆ# géns^gmensénék - lisá&en, 'V^:sp^b^ktaftènè->rh^ij^Â, amelyek műtéti útdn kapcsoltak az emltétf második VA-JA-CA génkíaszter egy érintetlen egér CA konstans régiböA éa ahoí áz Enb 2.4, egér lambda 3' errbancér (Enh) és Énb 1.1 endogén edbendefak érlniétlensk máradnák sz említett Λ kőrmyiiánc alíilben,
- 17. Mödszerr antitest léttdimzlsérs Rétben ; arrteiy tertsimazza: 8) egér kitételét egy antigénnek az 1-es igénypont szerint; b| az egér âiteli, az àbl^é«nÉ.:àdbii«bvàl^'.^iôdÔ8êRôk lehetővé tételéi és c) : égy antitest leválasztását a (b) szerinti egérből, amely kifejezetten az antigént Ismeri fel, vagy égy olyan immunoglobulin dómént tartalmazd sejt leválasztásét a (Öj szerinti egérből, amely klfejézétten az antigént isméd fel. vagy égy olyan nehéztánmls/vsgy konnyüSéns variábilis dömént.kálÉ^tby^l^y^Kvön^l f&É -szefW egérben, amely kötődik az antigénhez, ahol az antitest iegy hVA, tCA és ogy CA egér génből származiatött könnyőíáneaí tartalmaz.
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