Methods of making heavy chain only antibodies using transgenic animals
The present invention relates to a method of generation of fully functional heavy chain-only antibodies in transgenic mice in response to antigen challenge.
1. A method for the production of soluble, antigen-specific heavy chain only antibodies comprising:
(a) immunising a transgenic rodent expressing a heterologous V H heavy chain locus with an antigen wherein:
(i) the V H heavy chain locus comprises a variable region comprising at least one V H gene segment, from 20 to 40 D gene segments, at least one J gene segment and a heavy chain constant region comprising at least one constant region gene that does not encode a functional C H 1 domain;
(ii) a V H gene segment, a D gene segment and a J gene segment are capable of recombining to form a VDJ coding sequence;
(iii) the recombined V H heavy chain locus, when expressed upon antigen challenge, is capable of forming a soluble, heavy chain-only antibody comprising a soluble, antigen-specific V H binding domain and a constant effector region devoid of a functional C H 1 domain;
(b) cloning said recombined V H heavy chain locus from an antibody-producing cell of said immunised transgenic rodent after affinity maturation via somatic mutation; and
(c) producing said soluble, antigen specific heavy chain only antibody from the clone of step (b).
2. A method for the production of soluble, antigen-specific heavy chain only antibodies comprising:
(a) immunising a transgenic rodent expressing a heterologous heavy chain locus with an antigen wherein:
(i) the V H heavy chain locus comprises a variable region comprising at least one V H gene segment, from 20 to 40 D gene segments, at least one J gene segment and a heavy chain constant region comprising a Cγ or Cμ gene that does not encode a functional C H 1 domain;
(ii) a V H gene segment, a D gene segment and a J gene segment are capable of recombining to form a VDJ coding sequence;
(iii) the recombined V H heavy chain locus, when expressed upon antigen challenge, is capable of forming a soluble, heavy chain-only antibody comprising a soluble, antigen-specific V H binding domain and a Cγ or Cμ constant effector region devoid of a functional C H 1 domain;
(b) cloning said recombined V H heavy chain locus from an antibody-producing cell of said immunised transgenic rodent after affinity maturation via somatic mutation; and
(c) producing said soluble, antigen specific heavy chain only antibody from the clone of step (b).