Compound serum and hemin free medium for cultivation of Leishmania tarentolae: A recombinant protein expression system
AbstractSerum free cultivation of Leishmania is cost-effective and improves large scale production of well defined parasite material. Moreover, the production of recombinant pharmaceutical proteins requires cultivation of the host in a culture medium free of animal materials, so several culture media for Leishmania tarentolae expression system have been introduced. Some investigations have established the development of a serum-free, but hemin containing medium, based on yeast extract and buffer salts. Hemin is a substance of animal origin also interferes with nickel ions on Ni-NTA resin. In this study, L. tarentolae from Iranian lizard, cultivated in a compound serum and hemin free medium (LBR medium) and the growth parameters were determined. Here we report that LBR medium could obtain high maximal cell density of 1.8 × 108 cells ml-1 equivalent to that of hemin containing medium in our conditions. This compound medium was confirmed by successful expression of a 28 kDa his-tagged protein. With knowledge of the results, the easy-preparing culture medium could be used as a new culture medium for the production of recombinant proteins in L. tarentolae.
Keywords: Leishmania tarentolae, serum free cultivation of Leishmania, protein expression, Leishmania culture media.