Single domain ligands, receptors comprising said ligands, methods for their production, and use of said ligands and receptors
View Patent ↗The present invention relates to single domain ligands derived from molecules in the immunoglobulin (Ig) superfamily, receptors comprising at least one such ligand, methods for cloning, amplifying and expressing DNA sequences encoding such ligands, preferably using the polymerase chain reaction, methods for the use of said DNA sequences in the production of Ig-type molecules and said ligands or receptors, and the use of said ligands or receptors in therapy, diagnosis or catalysis.
1. A method for making an antibody to a target antigen which method comprises:
(a) generating an antibody variable domain expression library having a diversity of CDR3 sequences by a method comprising:
(i) providing expression vectors, said vectors comprising a variable domain encoding sequence of an antibody;
(ii) introducing by mutagenesis a diversity of CDR3 sequences into said variable domain encoding sequence;
(iii) recovering an expression library having a diversity of CDR3 binding activities; and
(iv) selecting a vector encoding an antibody variable domain having binding activity against the target antigen from said library;
(b) constructing a vector encoding an antibody comprising said selected antibody variable domain attached to an antibody chain constant domain; and
(c) expressing said antibody in a mammalian cell.
2. The method of claim 1 wherein said expression library having a diversity of CDR3 sequences is a VH domain library.
3. The method of claim 2 wherein said antibody chain constant domain is a heavy chain constant domain.
4. The method of claim 3 wherein said expressing in a mammalian cell is in association with a light chain to provide a complete antibody.
5. A method for making an antibody to a target antigen which method comprises
(a) providing an antibody variable domain encoding sequence selected from an expression library made by a method comprising:
(i) providing expression vectors, said vectors comprising a variable domain encoding sequence of an antibody;
(ii) introducing by mutagenesis a diversity of CDR3 sequences into said variable domain encoding sequence;
(iii) recovering an expression library having a diversity of CDR3 binding activities;
(b) constructing a vector encoding an antibody comprising said selected antibody variable domain encoding sequence attached to an antibody chain constant domain encoding sequence; and
(c) expressing said antibody in a mammalian cell.
6. A method for obtaining an antibody to a target antigen which method comprises:
(a) providing an expression library which expresses antibody variable domains, said library comprising a universal set of framework regions carrying a diversity of CDR3 sequences, said library having a diversity of binding activities,
(b) selecting from said library an antibody variable domain encoding sequence which encodes a variable domain which binds to said target antigen; and
(c) expressing said sequence attached to an antibody chain constant domain encoding region in a mammalian cell.
7. The method of claim 6 wherein said expression library having a diversity of CDR3 sequences is a VH library.
8. The method of claim 7 wherein said antibody chain constant domain is a heavy chain constant domain.
9. The method of claim 8 wherein said expressing in a mammalian cell is in association with a light chain to provide a complete antibody.