SUPPLEMENTATION OF GLYCINE ENHANCES THE EXPRESSION OF RECOMBINANT ANTI-TNFα SINGLE CHAIN VARIABLE FRAGMENT (SCFV) IN ESCHERICHIA COLIPREETI SAROHA, MOOKAMBESWARAN A. VIJAYALAKSHMI AND KRISHNAN VENKATARAMAN
Recently, there has been increased interest in the use of fragments of monoclonal antibodies for therapeutic purposes of treating autoimmune diseases, cancer etc, owing to its amenability for production using bacterial system. Anti-Tumour Necrosis Factor α (TNFα) single chain variable fragment (scFv) is being considered for the treatment of certain autoimmune diseases. To improve the production of anti-TNFα scFv in bacterial expression system, we report the usage of free amino acid or trace metal in nutrient rich media. We explored, Terrific Broth, 2x Yeast extract-Tryptone and Super Broth along with some additives (Magnesium Sulphate, Arginine and Glycine) to improve the expression of anti-TNFα scFv. The use of nutrient rich media increased the levels of its expression by 1.5 (Terrific Broth), 2.0 (2x Yeast extract- Tryptone) and 2.5 (Super Broth) folds, respectively. Inclusion of Magnesium Sulphate or Arginine in the media showed negligible expression of anti-TNFα scFv. Interestingly, supplementation of these media with glycine, significantly enhanced the expression of anti-TNFα scFv by 3.3 (Terrific Broth), 3.8 (2x Yeast extract- Tryptone) and 4.6 (Super Broth) folds respectively. We discuss the utility of Glycine and nutrient rich media for the production of recombinant anti-TNFα scFv.
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