Transgenic mice

Abstract

The invention relates to nucleic acid constructs for expression in mice for the reproduction of heavy chain only antibodies and V H domains, transgenic mice, related methods and uses.

1 . A vector comprising
 a) at least 10 functional human heavy chain V genes wherein at least 10 functional human heavy chain V genes are in their natural configuration;   b) at least one human heavy chain D gene and at least one human heavy chain J gene;   c) a murine C gene which lacks the C H 1 exon.   
     
     
         2 . A vector according to  claim 1  wherein said vector comprises a murine 3′ enhancer region. 
     
     
         3 . A vector according to  claim 1  wherein said vector comprises a murine 3′ enhancer region that is at least about 42 kb in size. 
     
     
         4 . A vector according to  claim 1  wherein said vector comprises a murine 3′ enhancer region and wherein said murine 3′ enhancer region comprises one or more enhancer elements selected from hs3A, hs1.2, hs3B, hs4, hs5, hs6 and hs7. 
     
     
         5 . (canceled) 
     
     
         6 . A vector according to  claim 1  wherein said vector comprises a murine μ enhancer. 
     
     
         7 . A vector according to  claim 1  wherein said vector comprises a switch μ element. 
     
     
         8 . A vector according to  claim 1  wherein said vector comprises a murine Cγ gene. 
     
     
         9 . A vector according to  claim 1  wherein said vector comprises a murine μ enhancer and a switch μ element or a murine μ enhancer and a murine Cγ gene. 
     
     
         10 . A vector according to  claim 1  wherein said vector comprises a murine Cγ 1 gene. 
     
     
         11 . A vector according to  claim 1  comprising 10 to about 44 functional human V genes. 
     
     
         12 . A vector according to  claim 1  further comprising at least one or more murine Cγ gene. 
     
     
         13 . A vector according to  claim 1  comprising a murine Cγ gene selected from Cγ2b and Cγ2a. 
     
     
         14 . A vector according to  claim 1  comprising Cγ2b and Cγ2a. 
     
     
         15 . A vector according to  claim 1  further comprising selection markers. 
     
     
         16 . A vector according to  claim 1  wherein said vector is a yeast artificial chromosome (YAC). 
     
     
         17 . A transgenic murine host cell transformed with a vector according to  claim 1  optionally wherein said cell is an ES cell. 
     
     
         18 . (canceled) 
     
     
         19 . A transgenic mouse comprising a vector according to  claim 1  or a cell transformed with a vector according to  claim 1 . 
     
     
         20 . A transgenic mouse according to  claim 19  wherein said mouse comprises one or more non-functional endogenous immunoglobulin loci. 
     
     
         21 . A transgenic mouse according to  claim 19  wherein said mouse comprises a non-functional endogenous lambda light chain locus and/or a non-functional endogenous kappa light chain locus and/or a non-functional endogenous heavy chain locus. 
     
     
         22 .- 27 . (canceled) 
     
     
         28 . A method for making a library using a transgenic mouse according to  claim 19  optionally wherein said library is a naïve library. 
     
     
         29 . (canceled) 
     
     
         30 . A method for making a library comprising ex vivo immunisation of a transgenic mouse according to  claim 19  or ex vivo tissue or cells of a transgenic mouse according to  claim 19 . 
     
     
         31 . (canceled) 
     
     
         32 . A method for making a HCAb or V H  domain comprising introducing and expressing a vector according to  claim 1  in a mouse or in a murine host cell. 
     
     
         33 . A method according to  claim 32  wherein said mouse comprises one or more non-functional endogenous immunoglobulin loci. 
     
     
         34 . A method according to  claim 33  wherein said mouse comprises a non-functional endogenous lambda light chain locus and/or a non-functional endogenous kappa light chain locus and/or a non-functional endogenous heavy chain locus. 
     
     
         35 .- 37 . (canceled) 
     
     
         38 . A method according to  claim 32  wherein said method comprises cloning the sequence encoding the V H  domain from mRNA derived from the isolated cell or tissue, constructing a library from cloned transcripts and isolating the V H  domain. 
     
     
         39 . A method for producing a soluble V H  binding domain comprising the following steps:
 a) expressing a vector of  claim 1  in a transgenic mouse,   b) isolating a cell or tissue expressing a HCAb,   c) cloning the sequence encoding the V H  domain from mRNA derived from the isolated cell or tissue,   d) constructing a library from cloned transcripts and   e) isolating the V H  domain.   
     
     
         40 . A soluble V H  domain obtained or obtainable by the method of  claim 39 . 
     
     
         41 . A composition comprising a V H  domain according to  claim 40  optionally in combination with another V H  domain, protein, or other molecule of therapeutic benefit. 
     
     
         42 . (canceled)