DNA LIBRARIES ENCODING FRAMEWORKS WITH SYNTHETIC CDR REGIONS

摘要

A synthetic DNA library or a member of a synthetic DNA library of antibodies or fragments of antibody molecules having heavy chain(s) and lacking light chain(s) is described wherein the CDR within variable domains are synthesized using randomly assembled trinucleotide phosphoramidites (trimer phosphoramidites) to eliminate unwanted cysteine amino acids and/or stop codons.

主权项

1. A method of preparing a synthetic single domain VHH library, comprising:
A. preparing synthetic sequences corresponding to cDNA of four framework regions (FR1-FR4) of a single domain VHH antibody, B. preparing synthetic sequences of three complementarity determining region sequences (CDR1-CDR3) of said VHH antibody by random assembly of nucleotide trimers wherein said synthetic sequences contain sequences complementary to the ends of adjacent framework regions for hybridization to said adjacent framework regions, and C. assembling a synthetic DNA library encoding VHH antibodies using said synthetic cDNA framework regions and said synthetic random nucleotide trimer sequences of said CDR regions with said complementary ends to said framework regions.