Binding molecules

摘要

The present invention relates to the manufacture of a diverse repertoire of functional heavy chain-only antibodies that undergo affinity maturation, and uses thereof. The invention also relates to the manufacture and use of a diverse repertoire of class-specific heavy chain-only antibodies and to the manufacture and use of multivalent polypeptide complexes with antibody heavy chain functionality, preferably antibody heavy chain binding functionality, constant region effector activity and, optionally, additional effector functions.;The present invention also relates to a method of generation of fully functional heavy chain-only antibodies in transgenic mice in response to antigen challenge. In particular, the present invention relates to a method for the generation of human antigen-specific, high affinity, heavy chain-only antibodies of any class, or mixture of classes and the isolation and expression of fully functional VH antigen-binding domains.

主权项

1. A method for the production of soluble, antigen-specific heavy chain only antibodies comprising:
(a) immunising a transgenic mouse expressing a heterologous VH heavy chain locus with an antigen wherein:
(i) the VH heavy chain locus comprises a variable region comprising at least one naturally occurring human VH gene segment, at least one D gene segment, at least one J gene segment and at least one heavy chain constant region;(ii) each constant region does not encode a functional CH1 domain;(iii) the VH gene segment, D gene segment and J gene segment are capable of recombining to form a VDJ coding sequence;(iv) the recombined VH heavy chain locus, when expressed upon antigen challenge, is capable of forming a soluble, heavy chain-only antibody comprising a soluble, antigen-specific VH binding domain and a constant effector region devoid of a functional CH1 domain with an antigen; (b) cloning a soluble heavy chain only antibody resulting from recombination between single VH, D and J gene segments and encoding a soluble, antigen-specific VH binding domain from an antibody-producing cell of said immunised transgenic mouse after affinity maturation via somatic mutation; and (c) producing said soluble, antigen specific heavy chain only antibody, or a soluble antigen-specific binding domain derived therefrom, from the clone of step (b).