"Preparation and use of gene banks of synthetic human antibodies (""synthetic human-antibody libraries"")"

US5840479A · kind A · utility

Assignee

• Behring Diagnostics GmbH · DE

Inventors

• Melvyn Little · Büdelsdorf, DE
• Frank Breitling · Heidelberg, DE
• Thomas Seehaus · Heppenheim, DE
• Stefan Dubel · Dossenheim, DE
• Iris Klewinghaus · Mannheim, DE

Key dates

• Filing date: Dec 2, 1994
• Grant date: Nov 24, 1998
• Expiry date: Dec 2, 2014

Classification

• Technology area (CPC C): Chemistry; Metallurgy
• CPC primary: C07K2319/033
• WIPO field: Biotechnology
• WIPO sector: Chemistry

Abstract

The invention relates to the preparation and use of gene banks of synthetic human antibodies (huAb) or parts of antibodies which contain the antigen-binding domain. Starting from a huAb framework in a suitable vector, the hypervariable regions of the antibody cDNA are formed by almost "randomly" combined oligonucleotides. Relatively conserved amino acids in the hypervariable regions have here been taken account of in the choice of appropriate nucleotides during the oligonucleotide synthesis and the ratio of the nucleotides used is likewise chosen such that a nonsense codon is to be expected at most in every 89th position. Expression of this synthetic huAb cDNA in microbial expression systems, e.g. in E. coli in the vector pFMT which is described below, thus makes a synthetic huAb library with a comprehensive repertoire for screening using selected antigens available in vitro.