Deutsches Krebsforschungszentrum Stiftung Des Offentlichen Rechts

Other

Dade Behring Marburg Gmbh

Behring Diagnostics Gmbh

Siemens Healthcare Diagnostics Products Gmbh

Iris Klewinghaus

Melvyn Little

Frank Breitling

Stefan Dubel

Stefan Dübel

Michael Braunagel

Stefan D&uuml;bel

Thomas Seehaus

Frank Berthold Breitling

A phagemid has been constructed that expresses an antibody fused to coliphage pIII protein. The phagemid is suitable for selecting specific antibodies from large gene libraries with small quantities of antigen. The antibody-pIII gene can be strongly repressed, so that it allows antibody libraries to be amplified without the danger of deletion mutants predominating. After induction, large quantities of the fusion protein may be expressed.

Phagemid for antibody screening

The invention relates to the preparation and use of gene banks of human antibodies (Ab). Starting from a mixture of human B-lymphocytes, their mRNA is translated into the cDNA using oligo-dT primers. Subsequently, an amplification of the Ab-specific cDNA by means of polymerase chain reaction (PCR) takes place using suitable oligonucleotide primer sequences. Expression of this amplified Ab-specific cDNA in a bacterial expression vector, e.g. the vector pFMT described below, in,thus makes available a human-antibody library with a comprehensive repertoire for screening selected antigens in vitro.

Preparation and use of gene banks of human antibodies (&ldquo;human-antibody libraries&rdquo;)

Phagemid library for antibody screening

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.

Preparation and use of gene banks of synthetic human antibodies

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Mannheim, Germany

Iris Klewinghaus