Recombinant vaccine protein

A. Purification of Proteins on MA Case Study of Recombinant Vaccine Protein... 

FIGURE 10 Optimization of elution salt step for a recombinant vaccine protein purification. A step elution ladder with increments of 100 m M salt for 5 min durations, from 0 to I /VI was used MA, Sartobind Q-15 (anionic 100 cm2). Load conditions double load, 15 m/VI Tris, pH 8.1 flow rate 5 mL/ min. Most of the vaccine protein elutes in the 200 m/VI salt step. FT indicates the flow through peaks. (Courtesy of R. McMaster, Aventis Pasteur.)... 

FIGURE I I Optimization of pH elution steps for recombinant vaccine protein purification. Incremental pH step gradients 5.0, 5.5, 6.0, 6.5, and 8.0 are shown. Membrane 100 cm2 C (Sartobind Cl00) Load conditions 15 m/VI citrate, pH 4.75, flow rate 7 mL/min. The product elutes at pH shift of 5.5. (Courtesy of R. McMaster, Aventis Pasteur.)... 

A particle that spontaneously assembles from viral coat proteins in the absence of other viral components. Vims-like particles (VLPs) generated from recombinant coat proteins are used in vaccines against hepatitis B vims and papillomavirus. 

PI. polymorpha [80] is generally referred to as safe and was widely used for pharma-protein production (cytokines, vaccines, coagulation factors) [80]. The genome sequence is known and DNA chip technologies have been established. Hansenula was developed to an expression platform which was implemented for the production of recombinant vaccines. 

Yeast expression vectors have been among those most commonly used since the beginning of gene technology. Vectors based on baker s yeast, Saccharomyces cerevisiae, have been especially popular for robust expression of many types of recombinant proteins [90]. For instance, the first commercially available recombinant vaccine, the hepatitis B surface antigen vaccine, was produced from an S. cerevisiae vector [91]. Many other recombinant proteins have also been efficiently expressed in yeast including al-Antitrypsin [92], insulin [93], Epstein-Barr virus envelope protein [94], superoxide dismutase [95] and interferon-a [90]. 

Gardasil is a noninfectious recombinant vaccine consisting of capsid proteins from four different human papillomaviruses (HPVs) of types 6,11, 16, and 18. HPV causes squamous cell cervical cancer and cervical adenocarcinoma, as well as 35-50% of vulvar and vaginal cancers. 

Intact plants are also suitable for cost-effective production of recombinant pharmaceutical proteins providing products free of contaminations with endotoxins or human pathogens. Plants generally promote the proper fold of foreign proteins and post-translational modifications that are somehow similar to those of mammalian systems. Also, production of vaccine candidates in cereal seeds allows antigen protection from proteolysis which ensures their stability for a long period of time. 

Mammalian cells are commonly employed for the production of therapeutic and diagnostic proteins, since they are able to correctly synthetize the large and complex structures that the human body requires as medicine [1]. Nowadays, they are employed for the large-scale production of recombinant therapeutic proteins, monoclonal antibodies (MAbs) and viruses used in the preparation of vaccines (e.g. against rabies, hepathytis B, polio, etc) [2]. An overview of some licensed/approved products derived from mammalian cell culture is given in Table 1. 

Pogrebnyak, N., Golovkin, M., Andrianov, V., Spitsin, S., Smirnov, Y, Egolf, R., and Koprowski, H. (2005). Severe acute respiratory syndrome (SARS) S protein production in plants development of recombinant vaccine. Proc. Natl. Acad. Sci. U.S.A. 102(25) 9062-9067. 

Lebens, M., Sun, J.B., Sadeghi, H., Backstrom, M., Olsson, I., Mielcarek, N., Li, B.L., Capron, A., Czerkinsky, C. and Holmgren, J. (2003) A mucosally administered recombinant fusion protein vaccine against schistosomiasis protecting against immunopathology and infection. Vaccine 21, 514-520. 

Georgiou, G., Stathopoulus, C., Daugherty, P. S., Nayak, A. R., Iverson, B. L., and Curtiss III, R. (1997). Display of heterologous proteins on the surface of microorganisms from the screening of combinatorial libraries to live recombinant vaccines. Nature Biotechnology, 15, 29-34. 

S6 outlines the products for which this guidance is applicable, such as recombinant DNA proteins, vaccines, peptides, plasma derived products, endogenous proteins extracted from human tissues or oligonucleotide drugs etc., while heparin, vitamins, and cellular blood components, for example, are not covered. 

Clinical studies with a recombinant vaccine (using Vaccinia virus expressing HPV 16, 18, E6, and E7 proteins) in patients with preinvasive and invasive cancer have been reviewed (1). 

A new recombinant vaccine has shown promise in vitro and in rabbits. It combines segments of 21 different immunogenic peptides from P. falciparum into a single recombinant protein, ehciting a multilayered immune response, including B cell and T cell responses (1). 

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