CPC C07K 16/005 (2013.01) [C07K 16/32 (2013.01); C07K 16/40 (2013.01); C07K 16/44 (2013.01); C40B 40/08 (2013.01); C07K 2317/22 (2013.01); C07K 2317/24 (2013.01); C07K 2317/565 (2013.01); C07K 2317/569 (2013.01); C07K 2317/76 (2013.01); C07K 2317/80 (2013.01); C07K 2317/92 (2013.01); C40B 40/10 (2013.01); C40B 50/06 (2013.01)] | 4 Claims |
1. A method of making a synthetic single domain antibody library, said method comprising
i. introducing a diversity of nucleic acids encoding CDR1, CDR2, and CDR3, between the respective framework coding regions of a synthetic single domain antibody to generate nucleic acids encoding a diversity of synthetic single domain antibodies with the same synthetic single domain antibody scaffold amino acid sequence,
wherein said synthetic single domain antibody scaffold amino acid sequence contains at least the following amino acid residues: F37, E44, R45, F47, and; at least the following amino acid residues: P15, S49, S81, R93, A94, and optionally further comprising the residues Q8, Q108 and T99, wherein the positions of amino acid residues are indicated according to the Kabat nomenclature used for VH amino acid sequence,
wherein said synthetic single domain antibody scaffold comprises the following framework regions consisting of FR1 of SEQ ID NO:1, FR2 of SEQ ID NO:2, FR3 of SEQ ID NO: 3 and FR4 of SEQ ID NO:4, with 1, 2 or 3 amino acid substitutions within one or more of the framework regions FR1-FR4.
|