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AssigneeDade Behring Marburg GmbH
InventorsMelvyn Little, Frank Berthold Breitling, Thomas Seehaus, Stefan Dübel, Iris Klewinghaus
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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 E. coli thus makes available a human-antibody library with a comprehensive repertoire for screening selected antigens in vitro.
A human-antibody library obtained by means of transcription of isolated mRNA from peripheral non-activated human B-lymphocytes into cDNA, subsequent amplification of cDNA coding for antibodies by polymerase chain reaction (PCR) by means of suitable IgM-specific primers, and subsequent incorporation in suitable expression plasmids and finally expression, in individual clones, of the relevant antibody RNA which is contained therein and has been amplified in cDNA.
A process for preparing a human-antibody library, which comprises mRNA being isolated from non-activated peripheral human B-lymphocytes and being transcribed into cDNA, subsequently amplifying the cDNA coding for antibodies by PCR by means of suitable IgM-specific primers, then carrying out an incorporation into suitable expression plasmids and finally expressing the antibody cDNA in individual clones.
A human antibody library comprising recombinant DNA molecules coding for antibodies wherein said DNA molecules are obtained by means of transcription of isolated mRNA from non-activated peripheral human B lymphocytes into cDNA, amplification of said cDNA by polymerase chain reaction using IgM specific primers and incorporation into a suitable expression vector.
A method of obtaining a human antibody library comprising:
The antibody expression plasmid pFMT.
Dependencies are inferred from the claim text.