WO2002028903A2 - Heavy chain libraries - Google Patents

Heavy chain libraries Download PDF

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Publication number
WO2002028903A2
WO2002028903A2 PCT/NL2001/000670 NL0100670W WO0228903A2 WO 2002028903 A2 WO2002028903 A2 WO 2002028903A2 NL 0100670 W NL0100670 W NL 0100670W WO 0228903 A2 WO0228903 A2 WO 0228903A2
Authority
WO
WIPO (PCT)
Prior art keywords
heavy chain
nucleic acid
chain variable
variable fragment
library
Prior art date
Application number
PCT/NL2001/000670
Other languages
French (fr)
Other versions
WO2002028903A3 (en
Inventor
Ton Logtenberg
Erwin Houtzager
Original Assignee
Crucell Holland B.V.
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Priority claimed from EP00203216A external-priority patent/EP1188771A1/en
Application filed by Crucell Holland B.V. filed Critical Crucell Holland B.V.
Priority to AU2001294399A priority Critical patent/AU2001294399A1/en
Priority to CA002421960A priority patent/CA2421960A1/en
Publication of WO2002028903A2 publication Critical patent/WO2002028903A2/en
Publication of WO2002028903A3 publication Critical patent/WO2002028903A3/en
Priority to US10/382,361 priority patent/US20030219829A1/en

Links

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
    • 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
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/20—Immunoglobulins specific features characterized by taxonomic origin
    • C07K2317/21—Immunoglobulins specific features characterized by taxonomic origin from primates, e.g. man
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/50—Immunoglobulins specific features characterized by immunoglobulin fragments
    • C07K2317/56—Immunoglobulins specific features characterized by immunoglobulin fragments variable (Fv) region, i.e. VH and/or VL

Abstract

The invention provides libraries which comprise binding molecules that are adapted to expression in an expression organism, but which are also transferable to a human context without undergoing a change in conformation and/or build up. The invention therefore provides a method for producing a human monoclonal antibody, said method comprising: providing a library of binding molecules, the binding domain of which consists essentially of human heavy chain variable fragments in a functional format, selecting from said library at least one heavy chain variable fragment having a desired binding affinity, and inserting a nucleic acid encoding said heavy chain variable fragment into a nucleic acid encoding the complementary part of at least a heavy chain of said human monoclonal antibody, allowing for expression of the resulting heavy chain and for assembly of said heavy chain with a desired light chain, and producing a human monoclonal antibody. The conformation of said heavy chain variable fragment is such that it retains its binding affinity whether it is in phage display or in its normal heavy chain environment. A method for making a library for use in said method is also herewith provided, as well as methods of keeping heavy chain variable fragments in said conformation. The present invention allows for producing larger libraries than the known ones. Besides, loss of specificities and affinities because of expression problems are reduced.

Claims

13Claims
1. A method for producing a human monoclonal antibody, said method comprising: providing a library of binding molecules, the binding domain of which consists essentially of human heavy chain variable fragments in a functional format, selecting from said library at least one heavy chain variable fragment having a desired binding affinity, and inserting a nucleic acid encoding said heavy chain variable fragment into a nucleic acid encoding the complementary part of at least a heavy chain of said human monoclonal antibody, allowing for expression of the resulting heavy chain and for assembly of said heavy chain with a desired light chain, and producing a human monoclonal antibody.
2. The method according to claim 1 wherein said heavy chain variable fragment is in a functional format through fusion to a structural protein designed for that purpose.
3. The method according to claim 1, wherein at least one sequence of said heavy chain variable fragment relevant only for association with a light chain is removed.
4. The method according to claim 1, wherein the complementary part of the heavy chain is derived from VH3, VH4 or VHl.
5. The method according to claim 1, wherein the light chain is derived from to a member of a Vkappal, Vkappa3 and VLambda3 gene family. 14
6. A human monoclonal antibody obtainable by a method according to claims 1-5.
7. A method for producing a structural amino acid sequence or a nucleic acid sequence encoding such an amino acid sequence for keeping a human heavy chain variable fragment in a functional format upon expression of a nucleic acid encoding such a fragment in a fusion with a nucleic acid encoding a protein expressed associated with the surface of a phage particle, said method comprising: fusing a nucleic acid sequence encoding a possible structural amino acid sequence to a nucleic acid which is a fusion of a human heavy chain variable fragment with a known binding affinity and said nucleic acid encoding a protein expressed associated with the surface of a phage particle, and expressing said nucleic acid in the context of a suitable phage expression system and selecting fusions which expose the desired binding affinity.
8. A proteinaceous substance or a nucleic acid encoding it, which substance is capable of keeping a heavy chain variable fragment in a functional conformation, obtainable by a method according to claim 7.
9. A method for making a library for use in a method according to claim 1, said method comprising: cloning a number of randomized nucleic acids derived from a heavy chain variable fragment in functional alignment with a nucleic acid encoding a proteinaceous substance according to claim 8, and providing the resulting nucleic acid in functional alignment with a nucleic acid encoding a protein expressed associated with the surface of a phage particle and expressing the resulting nucleic acids comprising 15 said heavy chain variable fragment, said proteinaceous substance encoding acid and said surface protein encoding nucleic acid in the context of a suitable phage expression system, thus producing said library.
10. A phage display library obtainable by a method according to claim 9.
PCT/NL2001/000670 2000-09-13 2001-09-12 Heavy chain libraries WO2002028903A2 (en)

Priority Applications (3)

Application Number Priority Date Filing Date Title
AU2001294399A AU2001294399A1 (en) 2000-09-13 2001-09-12 Heavy chain libraries
CA002421960A CA2421960A1 (en) 2000-09-13 2001-09-12 Heavy chain libraries
US10/382,361 US20030219829A1 (en) 2000-09-15 2003-03-05 Heavy chain libraries

Applications Claiming Priority (4)

Application Number Priority Date Filing Date Title
US23219200P 2000-09-13 2000-09-13
US60/232,192 2000-09-13
EP00203216A EP1188771A1 (en) 2000-09-15 2000-09-15 Libraries of human heavy chain variable fragments in a functional format
EP00203216.7 2000-09-15

Related Child Applications (1)

Application Number Title Priority Date Filing Date
US10/382,361 Continuation US20030219829A1 (en) 2000-09-15 2003-03-05 Heavy chain libraries

Publications (2)

Publication Number Publication Date
WO2002028903A2 true WO2002028903A2 (en) 2002-04-11
WO2002028903A3 WO2002028903A3 (en) 2003-01-23

Family

ID=26072693

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/NL2001/000670 WO2002028903A2 (en) 2000-09-13 2001-09-12 Heavy chain libraries

Country Status (3)

Country Link
AU (1) AU2001294399A1 (en)
CA (1) CA2421960A1 (en)
WO (1) WO2002028903A2 (en)

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP1345967B1 (en) * 2000-12-22 2011-08-10 Grad, Carole, Legal representative of Kaplan, Howard Phage display libraries of human v h fragments

Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1997002479A2 (en) * 1995-06-30 1997-01-23 Yale University Human monoclonal anti-tumor antibodies

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1997002479A2 (en) * 1995-06-30 1997-01-23 Yale University Human monoclonal anti-tumor antibodies

Non-Patent Citations (3)

* Cited by examiner, † Cited by third party
Title
J. THOMPSON ET AL.: "Affinity maturation of a high-affinity human monoclonal antibody against the third hypervariable loop of human immunodeficiency virus: Use of phage display to improve affinity and broaden strain reactivity." JOURNAL OF MOLECULAR BIOLOGY, vol. 256, no. 1, 16 February 1996 (1996-02-16), pages 77-88, XP002159769 London, GB *
P. SANNA ET AL.: "Directed selection of recombinant human monoclonal antibodies to herpes simplex virus glycoproteins from phage display libraries." PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE U.S.A., vol. 92, no. 14, 3 July 1995 (1995-07-03), pages 6439-6443, XP002159770 Washington, DC, USA *
R. POWELL ET AL.: "Construction, assembly and selection of combinatorial antibody libraries." CURRENT INNOVATIONS IN MOLECULAR BIOLOGY, vol. 5 (Genetic engineering with PCR), 1998, pages 155-172, XP000981992 Wymondham, GB *

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP1345967B1 (en) * 2000-12-22 2011-08-10 Grad, Carole, Legal representative of Kaplan, Howard Phage display libraries of human v h fragments

Also Published As

Publication number Publication date
CA2421960A1 (en) 2002-04-11
WO2002028903A3 (en) 2003-01-23
AU2001294399A1 (en) 2002-04-15

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