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40 The arrival of recombinant insulin in the pharmaceutical market paved a new direction for the clinical potential of proteins. Thus, the pharmaceutical industry underwent a paradigm shift towards proteins as therapeutic moieties. A number of recombinant protein drugs are available and many more are in pipeline to target major diseases, such as cancer, viral diseases, cardiovascular diseases and endocrine diseases [1]. In many cases, the recombinant protein is effective and plays an irreplaceable role in the treatment of the pathological condition. Such as hemophilia, except blood coagulation factor, which has an extremely limited source, the treatment mostly relies on the recombinant coagulation factor [2]. Many recombinant proteins are in clinical practice for more than two decades with an optimal safety and absence of side effects. As in case of growth hormone (GH) therapy, where administration of the growth hormone in patients with severe GH deficiency and hypopituitarism was found to be safe for longand short-term usage [3,4]. In terms of cancer protein therapeutics, currently only monoclonal antibodies, targeting over-expressed receptors, are prominently available for treatment of different tumors. The development of hybridoma technology for monoclonal antibody production revolutionized the treatment regime for solid tumors. Following approval of the first monoclonal antibody, rituximab, for clinical treatment of non-Hodgkin’s lymphoma, a number of monoclonal antibodies have been generated and are at present in clinical use for treatment of solid tumors [5,6]. However, the clinical success and the long-term benefits of recombinant protein therapy for treatment of several diseases have fostered enormous interest in their application for treatment of cancer. Li and colleagues reported anti-tumor effect of novaferon, a novel recombinant protein produced by DNA shuffling of interferon-α [7]. Evaluation of novaferon bioactivity on tumor cells displayed reduction in proliferation of HepG2 cells in vitro and in vivo. This promising pre-clinical study has provided strong lead for clinical trial verification. Likewise, Zhang and colleagues demonstrated HER2 receptor targeted activity of another recombinant protein e23sFv-Fdt-casp6 (immune-caspasse-6) in vitro and in vivo [8]. The recombinant protein e23sFv-Fdtcasp6 displayed inhibition of proliferation of HER-2 over-expressing A172 and U251MG in vitro, but not U-87 MG cells with undetectable HER-2 expression. Thus, the ability to produce targeted cytotoxicity makes the recombinant e23sFv-Fdt-casp6 a promising therapeutic alternative for Glioblastoma multiforme (GBM) treatment.

Protein Therapeutics from Monolayer to Spheroids- A Model for Preclinical Investigations