DE98200770T1 - Siebtestverfahren für Genbanken (recombinant libraries) - Google Patents

Siebtestverfahren für Genbanken (recombinant libraries) Download PDF

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Publication number
DE98200770T1
DE98200770T1 DE98200770T DE98200770T DE98200770T1 DE 98200770 T1 DE98200770 T1 DE 98200770T1 DE 98200770 T DE98200770 T DE 98200770T DE 98200770 T DE98200770 T DE 98200770T DE 98200770 T1 DE98200770 T1 DE 98200770T1
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Prior art keywords
heavy
antibody
light chain
bacteriophage
interest
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DE98200770T
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DE29800770U1 (de
Inventor
William J. Menlo Park Dower
Steven E. Palo Alto Cwirla
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Alere San Diego Inc
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Biosite Inc
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First worldwide family litigation filed litigation Critical https://patents.darts-ip.com/?family=24060691&utm_source=google_patent&utm_medium=platform_link&utm_campaign=public_patent_search&patent=DE98200770(T1) "Global patent litigation dataset” by Darts-ip is licensed under a Creative Commons Attribution 4.0 International License.
Application filed by Biosite Inc filed Critical Biosite Inc
Publication of DE98200770T1 publication Critical patent/DE98200770T1/de
Pending legal-status Critical Current

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Classifications

    • C—CHEMISTRY; METALLURGY
    • C40—COMBINATORIAL TECHNOLOGY
    • C40B—COMBINATORIAL CHEMISTRY; LIBRARIES, e.g. CHEMICAL LIBRARIES
    • C40B40/00—Libraries per se, e.g. arrays, mixtures
    • C40B40/02—Libraries contained in or displayed by microorganisms, e.g. bacteria or animal cells; Libraries contained in or displayed by vectors, e.g. plasmids; Libraries containing only microorganisms or vectors
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
    • C07K16/005—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies constructed by phage libraries
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/10—Processes for the isolation, preparation or purification of DNA or RNA
    • C12N15/1034—Isolating an individual clone by screening libraries
    • C12N15/1037—Screening libraries presented on the surface of microorganisms, e.g. phage display, E. coli display

Abstract

Verfahren zur Herstellung eines Antikörpers oder eines bindenden Fragments eines Antikörpers, umfassend
(a) Bakteriophagen-Vektortransformation einer Wirtszelle mit (i) einer ersten Nukleodidsequenz einer DNA-Bibliothek, die mit einer Sequenz fusioniert ist, welche ein Hüllprotein des Bakteriophagen codiert, und (ii) einer zweiten Nukleotidsequenz dieser Bibliothek, die mit einer Sequenz fusioniert ist, welche ein Signalpeptid codiert, so dass die Wirtszelle mit Nukleotidsequenzen transformiert wird, welche die variablen Regionen sowohl der schweren als auch der leichten Kette(n) codieren;
(b) Kultivierung der transformierten Wirtszelle unter Bedingungen, die geeignet sind zur Expression und zum Zusammenbau von Bakteriophagenpartikeln, die auf ihrer äußeren Oberfläche eine Bibliothek von mehrkettigen Proteinen präsentieren, welche die variablen Regionen der schweren und leichten Kette(n) umfassen;
(c) Selektion von Bakteriophagenpartikeln, die die interessierenden variablen Regionen der schweren und leichten Kette(n), welche spezifisch an einen Bindungspartner von Interesse binden, codieren, aus der Bibliothek;
(d) Umklonierung von Nukleinsäuren, welche die interessierenden variablen Regionen...

Claims (15)

  1. Verfahren zur Herstellung eines Antikörpers oder eines bindenden Fragments eines Antikörpers, umfassend (a) Bakteriophagen-Vektortransformation einer Wirtszelle mit (i) einer ersten Nukleodidsequenz einer DNA-Bibliothek, die mit einer Sequenz fusioniert ist, welche ein Hüllprotein des Bakteriophagen codiert, und (ii) einer zweiten Nukleotidsequenz dieser Bibliothek, die mit einer Sequenz fusioniert ist, welche ein Signalpeptid codiert, so dass die Wirtszelle mit Nukleotidsequenzen transformiert wird, welche die variablen Regionen sowohl der schweren als auch der leichten Kette(n) codieren; (b) Kultivierung der transformierten Wirtszelle unter Bedingungen, die geeignet sind zur Expression und zum Zusammenbau von Bakteriophagenpartikeln, die auf ihrer äußeren Oberfläche eine Bibliothek von mehrkettigen Proteinen präsentieren, welche die variablen Regionen der schweren und leichten Kette(n) umfassen; (c) Selektion von Bakteriophagenpartikeln, die die interessierenden variablen Regionen der schweren und leichten Kette(n), welche spezifisch an einen Bindungspartner von Interesse binden, codieren, aus der Bibliothek; (d) Umklonierung von Nukleinsäuren, welche die interessierenden variablen Regionen der schweren und leichten Kette(n) codieren, in einen Expressionsvektor; und (e) Herstellung des Antikörpers oder Antikörperfragments durch rekombinante Mittel.
  2. Verfahren gemäß Anspruch 1, wobei der Bakteriophagen-Vektor ein filamentöser Bakteriophage, vorzugsweise M13, fd oder f1, stärker bevorzugt fd oder ein Derivat davon ist.
  3. Verfahren gemäß Anspruch 1 oder Anspruch 2, worin das Hüllprotein pIII ist.
  4. Verfahren gemäß einem der Ansprüche 1 bis 3, worin das Signalpeptid ein Omp A, pel B, pho A, pIII oder β-Lactamase-Signalpeptid ist.
  5. Verfahren gemäß einem der Ansprüche 1 bis 4, bei dem die transformierten Wirtszellen nach der Kultivierung lysiert werden und die Bakteriophagenpartikel aus den zellulären Bruchstücken selektiert werden; wobei gegebenenfalls die Bakteriophagenpartikel, die das interessierende Protein codieren, dadurch angereichert werden, dass der Selektionsschritt wenigstens einmal wiederholt wird.
  6. Verfahren gemäß Anspruch 1, wobei die erwähnte erste Nukleotidsequenz der Bibliothek ein Protein codiert, das eine variable Region der schweren Kette(n) eines Antikörpers umfasst.
  7. Verfahren gemäß Anspruch 6, wobei sich diese variable Region der schweren Kette(n) des Antikörpers am Amino-Terminus des Hüllproteins auf der Bakteriophagenoberfläche befindet.
  8. Verfahren gemäß Anspruch 7, wobei das Hüllprotein ein vollständiges pIII Hüllprotein ist.
  9. Verfahren gemäß einem der Ansprüche 1 bis 8, bei dem die ersten und zweiten DNA-Sequenzen der Bibliothek amplifizierte cDNA umfassen.
  10. Verfahren gemäß einem der vorhergehenden Ansprüche, wobei der Expressionsvektor ein eukaryotischer Expressionsvektor oder ein prokaryotischer Expressionsvektor ist.
  11. Verfahren gemäß einem der vorhergehenden Ansprüche, wobei der Antikörper oder das Antikörperfragment die interessierenden variablen Regionen der schweren und leichten Kette(n) verbunden mit konstanten Regionen der schweren und leichten Kette(n) enthält.
  12. Verfahren gemäß einem der vorhergehenden Ansprüche, wobei der Antikörper oder das Antikörperfragment ein Fab Antikörperfragment ist, welches die interessierenden variablen Regionen der schweren und leichten Kette(n) enthält.
  13. Verfahren gemäß einem der vorhergehenden Ansprüche, wobei die Nukleotidsequenzen, die die interessierenden variablen Regionen der schweren und leichten Kette(n) codieren, nach mehreren Durchläufen der Schritte (a), (b) und (c) selektiert werden.
  14. Verfahren gemäß einem der vorhergehenden Ansprüche, wobei das Verfahren außerdem eine nach dem Zufallsprinzip ablaufende Rekombination von isolierten Sequenzen der variablen Regionen von schweren und leichten Ketten zur Transformation in die Wirtszelle umfasst.
  15. Verfahren gemäß einem der vorhergehenden Ansprüche, umfassend außerdem den Schritt der Inkorporierung des Antikörpers oder bindenden Antikörperfragments in eine therapeutische, prophylaktische oder diagnostische Zusammensetzung.
DE98200770T 1990-05-01 1991-05-01 Siebtestverfahren für Genbanken (recombinant libraries) Pending DE98200770T1 (de)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US517659 1990-05-01
US07/517,659 US5427908A (en) 1990-05-01 1990-05-01 Recombinant library screening methods

Publications (1)

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DE98200770T1 true DE98200770T1 (de) 2004-09-30

Family

ID=24060691

Family Applications (3)

Application Number Title Priority Date Filing Date
DE69130563T Revoked DE69130563T2 (de) 1990-05-01 1991-05-01 Screening-verfahren für genbanken (recombinant libraries)
DE69133437T Revoked DE69133437T2 (de) 1990-05-01 1991-05-01 Siebtestverfahren für Genbanken (recombinant libraries)
DE98200770T Pending DE98200770T1 (de) 1990-05-01 1991-05-01 Siebtestverfahren für Genbanken (recombinant libraries)

Family Applications Before (2)

Application Number Title Priority Date Filing Date
DE69130563T Revoked DE69130563T2 (de) 1990-05-01 1991-05-01 Screening-verfahren für genbanken (recombinant libraries)
DE69133437T Revoked DE69133437T2 (de) 1990-05-01 1991-05-01 Siebtestverfahren für Genbanken (recombinant libraries)

Country Status (10)

Country Link
US (2) US5427908A (de)
EP (3) EP0866136B1 (de)
JP (1) JP3344584B2 (de)
AT (2) ATE286985T1 (de)
AU (1) AU7793991A (de)
DE (3) DE69130563T2 (de)
DK (2) DK0527839T3 (de)
ES (2) ES2124224T3 (de)
GR (1) GR3029421T3 (de)
WO (1) WO1991017271A1 (de)

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US5427908A (en) 1995-06-27
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