US2014041067A1PendingUtilityA1

Antibodies, variable domains & chains tailored for human use

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Assignee: KYMAB LTDPriority: Sep 19, 2011Filed: Oct 11, 2013Published: Feb 6, 2014
Est. expirySep 19, 2031(~5.1 yrs left)· nominal 20-yr term from priority
Inventors:Allan BradleyGlenn FriedrichE-Chiang LeeMark StrivensNicholas England
C07K 2317/56C07K 16/462C07K 16/46A61P 31/00C12N 2800/204C07K 16/1232C07K 16/18A01K 2267/01C12N 15/8509C12N 15/85A61P 31/04C07K 2317/24A01K 2227/105A01K 2217/072A61P 31/22A61P 31/12C07K 16/1217A01K 67/0278C07K 16/088A61P 31/16A61P 33/06A01K 2217/15C07K 16/1242C07K 16/085A61P 31/18C07K 16/114C07K 16/108C07K 16/089C07K 16/12C07K 16/08
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Claims

Abstract

The invention relates to the provision of antibody therapeutics and prophylactics that are tailored specifically for human use. The present invention provides libraries, vertebrates and cells, such as transgenic mice or rats or transgenic mouse or rat cells. Furthermore, the invention relates to methods of using the vertebrates to isolate antibodies or nucleotide sequences encoding antibodies. Antibodies, heavy chains, polypeptides, nucleotide sequences, pharmaceutical compositions and uses are also provided by the invention.

Claims

exact text as granted — not AI-modified
1 . A non-human vertebrate whose genome comprises an immunoglobulin heavy chain locus comprising one or more VH gene segments, one or more D gene segments, and one or more J segments comprising human gene segment JH6*02, upstream of a constant region; wherein said gene segments in said heavy chain locus are operably linked to said constant region, and said vertebrate is functional to recombine a said VH gene segment, a said D gene segment and said human JH6*02 to form a VH-D-JH6*02 variable region gene. 
     
     
         2 . The vertebrate of  claim 1 , wherein said vertebrate has been immunised with a target antigen and comprises an antibody heavy chain comprising a variable domain encoded by said VH-D-JH6*02 gene produced by recombination of said heavy chain gene segments with said human JH6*02, wherein said variable domain comprises an HCDR3 having a length of at least 20 amino acids. 
     
     
         3 . A non-human vertebrate cell whose genome comprises an immunoglobulin heavy chain locus comprising one or more VH gene segments, one or more D gene segments and one or more J segments comprising human gene segment JH6*02, upstream of a constant region, wherein said gene segments of said heavy chain locus are operably linked to said constant region and said vertebrate is functional to recombine a said VH segment, a said D segment and said human JH6*02 gene segment to form a VH-D-JH6*02 variable region gene. 
     
     
         4 . The cell of  claim 3 , which is an ES cell, wherein said ES cell is capable of developing into a mouse comprising said immunoglobulin heavy chain locus in its germ line, and wherein said mouse comprises an antibody-producing cell that expresses an antibody comprising said heavy chain. 
     
     
         5 . The vertebrate of  claim 1 , wherein said heavy chain locus comprises a human JH6*02 recombination signal sequence (RSS) positioned upstream of and operably linked to said JH6*02 gene segment. 
     
     
         6 . The cell of  claim 3 , wherein the heavy chain locus comprises a human JH6*02 recombination signal sequence (RSS) positioned upstream of and operably linked to said JH6*02 gene segment. 
     
     
         7 . The vertebrate of  claim 5 , wherein said RSS is SEQ ID NO: 238 or, wherein said SEQ ID NO: 238 comprises a flanking 9mer, a central 22mer and a flanking 7mer, and said sequence comprises identical 9mer and 7mer flanking sequences and said central sequence is at least 70% identical to said 22mer central sequence of SEQ ID NO: 238. 
     
     
         8 . The cell of claim of  claim 6 , wherein the RSS is SEQ ID NO: 238 or, wherein said SEQ ID NO: 238 comprises a flanking 9mer, a central 22mer and a flanking 7mer, and said sequence comprises identical 9mer and 7mer flanking sequences and said central sequence is at least 70% identical to said 22mer central sequence of SEQ ID NO: 238. 
     
     
         9 . The vertebrate of  claim 6 , wherein said RSS and JH6*02 are provided as SEQ ID NO: 237. 
     
     
         10 . The cell of  claim 8 , wherein said RSS and JH6*02 are provided as SEQ ID NO: 237. 
     
     
         11 . The vertebrate of  claim 1 , wherein said JH6*02 is the only JH6 gene segment in the genome. 
     
     
         12 . The cell of  claim 3 , wherein said JH6*02 is the only JH6 gene segment in said genome. 
     
     
         13 . The vertebrate of  claim 1 , wherein said J gene segments comprise a plurality of J gene segments comprising a 3′ J gene segment positioned downstream in said plurality and upstream of said constant region in said locus, and said 3′ J gene segment comprises said JH6*02 gene segment. 
     
     
         14 . The cell of  claim 3 , wherein said J gene segments comprise a plurality of J gene segments comprising a 3′ J gene segment positioned downstream in said plurality and upstream of said constant region in said locus, and said 3′ J gene segment comprises said JH6*02 gene segment. 
     
     
         15 . The vertebrate of  claim 1 , wherein said locus comprises one, more or all human D gene segments D3-9; D4-17; D3-10; D2-2; D5-24; D6-19; D3-22; D6-13; D5-12; D1-26; D1-20; D5-18; D3-16; D2-21; D1-14; D7-27; D1-1; D6-25; D2-14; and D4-23. 
     
     
         16 . The cell of  claim 3 , wherein the locus comprises one, more or all human D gene segments D3-9; D4-17; D3-10; D2-2; D5-24; D6-19; D3-22; D6-13; D5-12; D1-26; D1-20; D5-18; D3-16; D2-21; D1-14; D7-27; D1-1; D6-25; D2-14; and D4-23. 
     
     
         17 . The vertebrate of  claim 15 , wherein the locus comprises one, more or all human D gene segments D3-9, D3-10, D6-19, D4-17, D6-13, D3-22, D2-2, D2-25 and D3-3. 
     
     
         18 . The cell of  claim 16 , wherein the locus comprises one, more or all human D gene segments D3-9, D3-10, D6-19, D4-17, D6-13, D3-22, D2-2, D2-25 and D3-3. 
     
     
         19 . The vertebrate of  claim 1 , wherein the locus comprises a plurality of human D gene segments and said JH6*02 is in human germline configuration with respect to the 3′-most human D gene segment. 
     
     
         20 . The cell of  claim 3 , wherein the locus comprises a plurality of human D gene segments and the JH6*02 is in human germline configuration with respect to the 3′-most human D gene segment. 
     
     
         21 . The vertebrate of  claim 1 , wherein the locus comprises one, more or all of IGHV gene segments selected from V3-21, V3-13, V3-7, V6-1, V1-8, V1-2, V7-4-1, V1-3, V1-18, V4-4, V3-9, V3-23, V3-11 and V3-20. 
     
     
         22 . The cell of  claim 3 , wherein the locus comprises one, more or all of IGHV gene segments selected from V3-21, V3-13, V3-7, V6-1, V1-8, V1-2, V7-4-1, V1-3, V1-18, V4-4, V3-9, V3-23, V3-11 and V3-20. 
     
     
         23 . The vertebrate of  claim 1 , wherein the locus comprises one, more or all of human D3-9*01, D3-10*01, D6-19*01, D6-13*01, D1-26*01, IGHV1-8*01, IGHV4-61*01, IGHV6-1*01, IGHV4-4*02, IGHV1-3*01, IGHV3-66*03, IGHV3-7*01 and IGHV3-9*01. 
     
     
         24 . The cell of  claim 3 , wherein the locus comprises one, more or all of human D3-9*01, D3-10*01, D6-19*01, D6-13*01, D1-26*01, IGHV1-8*01, IGHV4-61*01, IGHV6-1*01, IGHV4-4*02, IGHV1-3*01, IGHV3-66*03, IGHV3-7*01 and IGHV3-9*01. 
     
     
         25 . An antibody-producing cell that is a progeny of the cell of  claim 3 , wherein the antibody-producing cell comprises a heavy chain locus comprising a rearranged variable region produced by recombination of human JH6*02 with a D gene segment and a VH gene segment. 
     
     
         26 . The cell of  claim 25 , which is a B-cell or hybridoma that expresses a target antigen-specific antibody comprising a heavy chain that comprises a rearranged variable region encoded by a recombined Vh-D-human JH6*02 gene produced by recombination of said gene segments with said human JH6*02. 
     
     
         27 . The vertebrate of  claim 1 , wherein the antibody heavy chain specifically binds a target antigen. 
     
     
         28 . The cell of  claim 3 , wherein the antibody heavy chain specifically binds a target antigen. 
     
     
         29 . The vertebrate of  claim 1 , wherein said antibody heavy chain comprises a variable region comprising a HCDR3 length of at least 20 amino acids. 
     
     
         30 . The cell of  claim 3 , wherein said antibody heavy chain comprises a variable region comprising a HCDR3 length of at least 20 amino acids. 
     
     
         31 . The vertebrate of  claim 1 , wherein said antibody heavy chain is a product of the recombination of JH6*02 with a human VH gene segment recited in  claim 21  or  23  and/or a D gene segment recited in  claim 15 ,  17  or  23 . 
     
     
         32 . The cell of  claim 3 , wherein said antibody heavy chain is a product of the recombination of JH6*02 with a human VH gene segment recited in  claim 22  or  24  and/or a D gene segment recited in  claim 16 ,  18  or  24 . 
     
     
         33 . The vertebrate of  claim 1 , wherein said genome comprises endogenous heavy chain variable region gene segments which are nonfunctional to produce endogenous Ig heavy chain. 
     
     
         34 . The cell of  claim 3 , wherein said genome comprises endogenous heavy chain variable region gene segments which are nonfunctional to produce endogenous Ig heavy chain. 
     
     
         35 . The vertebrate of  claim 1 , wherein the genome is homozygous for said heavy chain locus. 
     
     
         36 . The cell of  claim 3 , wherein the genome is homozygous for said heavy chain locus. 
     
     
         37 . The vertebrate of  claim 1 , wherein the constant region is an endogenous constant region. 
     
     
         38 . The cell of  claim 3 , wherein the constant region is an endogenous constant region. 
     
     
         39 . A method for producing an antibody specific to a target antigen, the method comprising immunizing the non-human vertebrate according to  claim 1  with said antigen, thereby producing said antibody specific to said target antigen, wherein said antibody comprises a heavy chain variable region comprising an HCDR3, and wherein said heavy chain variable region is a product of the recombination of human JH6*02 with a VH and a D gene segment. 
     
     
         40 . The method of  claim 39 , further comprising the step of isolating said antibody. 
     
     
         41 . The method of  claim 40 , wherein the constant region is an endogenous constant region. 
     
     
         42 . A method for producing a human an antibody comprising carrying out the method of  claim 40  or  41 , wherein the constant region of the locus is a non-human vertebrate constant region, further comprising the step of replacing the non-human constant region of the isolated antibody with a human constant region. 
     
     
         43 . The method of  claim 39 , further comprising the step of isolating a B cell from said mouse, wherein said B cell expresses said antibody. 
     
     
         44 . The method of  claim 43 , further comprising the step of producing a hybridoma from said B cell. 
     
     
         45 . The method of  claim 44 , further comprising the step of isolating said antibody from said hybridoma. 
     
     
         46 . The method of  claim 39 , wherein the non-constant region is an endogenous constant region. 
     
     
         47 . The method of  claim 46 , further comprising the step of isolating a B cell from said mouse, wherein said B cell expresses said antibody. 
     
     
         48 . The method of  claim 47 , further comprising the step of producing a hybridoma from said B cell, wherein said hybridoma expresses said antibody. 
     
     
         49 . The method of  claim 48 , further comprising the step of isolating said antibody from said hybridoma. 
     
     
         50 . A method for producing a heavy chain or VH domain thereof, of an antibody specific to a target antigen, the method comprising immunizing a non-human vertebrate according to  claim 1  with the antigen, thereby producing said heavy chain or VH domain thereof, of an antibody specific to said target antigen, wherein the heavy chain or VH domain thereof comprises an HCDR3, and wherein said VH domain is a product of the recombination of human JH6*02 with a VH and a D gene segment. 
     
     
         51 . The method of  claim 50 , further comprising the step of isolating said heavy chain or VH domain thereof. 
     
     
         52 . The method of  claim 51 , wherein the constant region is an endogenous constant region. 
     
     
         53 . The method of  claim 50 ,  51  or  52 , wherein the heavy chain or VH domain thereof, comprises mouse AID-pattern somatic hypermutations and/or mouse dTd-pattern mutations. 
     
     
         54 . The method of  claim 39  or  46 , wherein the heavy chain or VH domain thereof, of said antibody comprises mouse AID-pattern somatic hypermutations and/or mouse dTd-pattern mutations. 
     
     
         55 . A method for producing a human an antibody comprising carrying out the method of  claim 40  or  41 , wherein the constant region of the locus is a non-human vertebrate constant region, wherein the heavy chain or VH domain of said antibody comprises mouse AID-pattern somatic hypermutations and/or mouse dTd-pattern mutations, further comprising the step of replacing the non-human constant region of the isolated antibody with a human constant region. 
     
     
         56 . The method of  claim 50 , further comprising the step of operably linking the VH domain with a human constant domain.

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