QUICK METHOD OF CLONING AND EXPRESSION OF RELATED ANTIBODY VARIABLE REGION GENE SEGMENTS Russian patent published in 2017 - IPC C07K16/00 C12N15/63 C12N15/10 

FIELD: biotechnology.

SUBSTANCE: method antibody production comprising antibody isolation from the eukaryotic cell culture medium consisting of nucleic acid encoding the antibody, wherein the encoding nucleic acid is obtained by amplification of nucleic acids encoding the relative variable domain, using the following as a template in a polymerase chain reaction (PCR): single stranded cDNA derived from RNA of antibody-secreting B cell, and nucleic acids encoding a variable domain, inserted into eukaryotic expression plasmid by ligation independent cloning, where a pool of nucleic acids encoding a variable domain of heavy and light antibody chains is used for insertion, where the B-cell is a single B-cell and where the B-cell and its progeny produce more than 20 ng/ml of antibodies for isolation of nucleic acids segments, encoding antibody variable domains for 7 days of their co-culturing with feeder cells, starting from a single cell; and introduction of the selected nucleic acids segments into eukaryotic expression plasmids is carried out without intermediate isolation and analysis of clonal intermediate plasmids.

EFFECT: according to the inventionm the method does not require intermediate cloning, selection and analysis of intermediate plasmids.

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