Production of recombinant HIV‐1 nef protein using different expression host systems: A techno‐economical comparison

Abstract

Three popular expression host systems Escherichia coli, Pichia pastoris and Drosophila S2 were analyzed techno-economically using HIV-1 Nef protein as the model product. On scale of 100 mg protein, the labor costs corresponded to 52-83% of the manufacturing costs. When analyzing the cost impact of the different phases (strain/cell line construction, bioreactor production, and primary purification), we found that with the microbial host systems the strain construction phase was most significant generating 56% (E. coli) and 72% (P. pastoris) of the manufacturing costs, whereas with the Drosophila S2 system the cell line construction and bioreactor production phases were equally significant (46 and 47% of the total costs, respectively). With different titers and production goal of 100 mg of Nef protein, the costs of P. pastoris and Drosophila S2 systems were about two and four times higher than the respective costs of the E. coli system. When equal titers and bioreactor working volumes (10 L) were assumed for all three systems, the manufacturing costs of the bioreactor production of the P. pastoris and Drosophila S2 systems were about two and 2.5 times higher than the respective costs of the E. coli system. V V C 2009 American Institute of Chemical Engineers Biotechnol. Prog., 25: 95-102, 2009

On-line measurement of the substrate concentrations in Pichia pastoris fermentations using FT-IR/ATR

“…and KM71H (phenotype Mut S ), and the product was recombinant HIV-1 Nef (negative factor) protein (Sirén et al 2006). The fermentation methodology is described in greater detail in Vermasvuori et al (2009). In the Mut S cultivation the temperature was kept constant at 30°C, whereas in the Mut ?…”

Rachiplusia nu larva as a biofactory to achieve high level expression of horseradish peroxidase

“…Even when processes based on insect cells are gaining ground rapidly among expression systems, the commercial production of recombinant proteins is limited because of their high cost. Vermasvuori et al (2009) compared HIV-1 Nef protein production using different host expression systems, and considered that the insect cell-based strategy was four times more expensive than that based on Escherichia coli, mainly due to the higher price of raw materials. A large investment in highly technical equipment (e.g.…”

Efficient and easily scalable protein folding strong anion exchange chromatography for renaturation and simultaneous purification of recombinant human asparaginase from E. coli

“…Recombinant human Asparaginase (rhASP) is a novel, best‐selling recombinant drug in the world. Recombinant proteins are generally expressed in E. coli as insoluble inclusion bodies for the production of therapeutic proteins . The process of purifying and recovering a protein from inclusion bodies is tedious.…”

“…Recombinant proteins are generally expressed in E. coli as insoluble inclusion bodies for the production of therapeutic proteins. 14 The process of purifying and recovering a protein from inclusion bodies is tedious. Chaotropes like Urea (8 mol/L) or guanidine-HCl (6 mol/L) are used in high concentrations to solubilize the target protein.…”