Converting rabbit hybridoma into recombinant antibodies with effective transient production in an optimized human expression system

Mouse monoclonal antibodies are standard in research and diagnostic. Among all advantages of mouse monoclonal antibodies there is a lack of affinity, sensitivity and specificity especially for small epitopes and antigens that are non-immunogenic to rodents. This suggests that a huge number of diseases are under-diagnosed because of non-availability of high-quality antibodies. Rabbit monoclonal antibodies (Rab-mabs) show 10 to 100 times higher affinities than mouse monoclonal antibodies and impress by improved specificity which is extremely important in all fields of clinical and non-clinical diagnostic. However, all advantages of Rab-mabs are eliminated by the low productivity of monoclonal rabbit hybridoma. To meet the needs of IVD manufacturers InVivo implemented a rapid Rab-mab production system by transient gene expression to generate mg to g scales of highly affine, sensitive and specific IgGs within weeks.

Successful conversion of Rab-mabs into recombinant rabbit antibodies requires a combination of different scientific techniques. Firstly needed is the preparation of cDNA and sequencing of the antibody variable regions out of the rabbit hybridoma cell line. Second, synthesized full length rabbit antibody cDNA are cloned into mammalian expression vectors. Third, recombinant rabbit antibody is produced in mammalian cells preserving high productivity at good viability and suitable cell density. InVivo has implemented a novel technology for efficient transient transfection and expression in HEK cells, being especially designed for high-yielding recombinant antibody production. In brief, we developed a low cytotoxicity transfection reagent and a culture medium that can be used for transfection and production. For the establishment of an optimized host cell line for transient gene expression processes we utilized directed evolution. This completed the production platform for high throughput approaches and large scale transfection for the production of gram quantities IgG within days.

Dr. Tim Welsink presented workflow, optimization of expression and case study data on producing recombinant Rab-mabs at the HAH conference in November 2016. Click here for the final presentation as pdf

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