Article

Production of therapeutic proteins with baculovirus expression system in insect cell

Institute of Biotechnology, Wonkwang University, Iksan, Korea
Entomological Research (Impact Factor: 0.4). 11/2008; 38(s1):S71 - S78. DOI: 10.1111/j.1748-5967.2008.00177.x

ABSTRACT

Recombinant DNA technology has a major advantage in that it is capable of producing specific therapeutic proteins on demand in a heterologous expression system. The extent of this notion can be understood when one considers how crucial such proteins are, and how problematic the economical and safe production of such proteins are. Therapeutic recombinant protein production is a fundamental aspect of 21st century biotechnology industries. The improved therapeutic recombinant protein expression systems that use prokaryotic and eukaryotic cells have enabled the development of a multi-billion dollar industry. Among the variety of available heterologous expression systems, the baculovirus-based insect cell expression system has been utilized frequently for the high-level production of therapeutic recombinant proteins. Thus, the baculovirus expression system has been recognized as one of the most powerful expression technologies for production, by virtue of the achievable amount and purity, and the ease of the eukaryotic production process. The majority of therapeutic proteins are glycoproteins originating from humans. The insect-based expression system harbors glycosylation processing pathways, which constitute an advantage over other prokaryotic systems that lack glycosylation. However, there are several drawbacks which must be circumvented in order to establish an efficient system for the production of recombinant proteins. This review presents a brief overview of the perspective, particularly the glycosylation aspect, of the production of therapeutic recombinant proteins via a baculovirus-based insect cell expression system.

Download full-text

Full-text

Available from: Arshad Jamal
  • Source
    • "In addition to an increase in productivity induced by growth arrest, cultivation of cells under mild hypothermic conditions offers other relevant advantages: extended culture times (lower cell populations reduce overall nutrient uptake and waste production) [24], decreased O2 demand [25], reduced intermolecular product aggregation [26], increased sensitivity to pH changes [27,28], and a decreased sensitivity to pro-apoptotic agents [29,30]. Protein sialylation [24], acidic glycoforms [31], and antennary structures [31] are relevant quality parameters that are improved in cells cultured under hypothermic conditions. Beyond to providing a good nutritional environment for cell growth, complete replacement of the culture medium provided an adequate culture environment for FVIII expression and maintenance of biological activity. "
    [Show abstract] [Hide abstract]
    ABSTRACT: Hemophilia A is a bleeding disorder caused by deficiency in coagulation factor VIII. Recombinant factor VIII (rFVIII) is an alternative to plasma-derived FVIII for the treatment of hemophilia A. However, commercial manufacturing of rFVIII products is inefficient and costly and is associated to high prices and product shortage, even in economically privileged countries. This situation may be solved by adopting more efficient production methods. Here, we evaluated the potential of transient transfection in producing rFVIII in serum-free suspension HEK 293 cell cultures and investigated the effects of different DNA concentration (0.4, 0.6 and 0.8 μg/106 cells) and repeated transfections done at 34° and 37 °C. We observed a decrease in cell growth when high DNA concentrations were used, but no significant differences in transfection efficiency and in the biological activity of the rFVIII were noticed. The best condition for rFVIII production was obtained with repeated transfections at 34 °C using 0.4 μg DNA/106 cells through which almost 50 IU of active rFVIII was produced six days post-transfection. Serum-free suspension transient transfection is thus a viable option for high-yield-rFVIII production. Work is in progress to further optimize the process and validate its scalability.
    Full-text · Article · Nov 2011 · BMC Biotechnology
  • [Show abstract] [Hide abstract]
    ABSTRACT: Monoclonal antibodies (Mabs) are biopharmaceuticals that are used increasingly for the treatment of a wide range of diseases such as cancer and autoimmunity. The effectiveness of therapeutic Mabs, most of which are immunoglobulin G (IgG), is dependent upon their ability to link antigen recognition with an appropriate effector function, to elicit a biological response in vivo that will treat the targeted disease. Studies over the last decade have determined that the effector function of Mabs is highly dependent upon the structure of the N-linked glycan of the Fc domain of the Mab. Total removal of the glycan is highly detrimental to the effector function of the Mab, but subtle differences in the glycan structure, such as the lack of fucose, can improve significantly bioactivity and function of the Mabs. Some Mabs are glycosylated in the variable Fab domain but in many cases the function is not known. The host cellular production system including the bioreactor environment can produce Mabs with very different glycosylation profiles that must be considered in bioprocess development. Cell culture conditions such as dissolved oxygen, nutrient levels, pH and feed strategies can all have considerable influence on the glycosylation of the Mab, which will affect product quality and efficacy. Great improvements have been made in techniques for high resolution and high throughput analysis of glycans such as normal phase-high performance liquid chromatography (HPLC) and mass spectrometry (MS). This has allowed a better understanding of the link between the structure and function, which will in turn lead to the development of safer and more effective Mabs. KeywordsAntibody-Glycosylation-Immunoglobulin-Glycan-CHO-Insect-Yeast
    No preview · Chapter · Jan 1970
  • [Show abstract] [Hide abstract]
    ABSTRACT: The baculovirus Autographa californica nuclear polyhedrosis virus is an attractive candidate for eukaryotic virus display. A variety of strategies exists to incorporate and present target proteins on the surface of infected insect cells as well as on budded virions. Native baculovirus proteins such as the major envelope protein and the capsid protein, but also foreign scaffolds such as the vesicular stomatitis virus G-1 protein or the influenza A virus hemagglutinin serve as fusion partners for surface presentation of target proteins. The purposes of surface display are manifold; efficient presentation of antigenic epitopes on budded virions serves to induce specific immune response, designed surface odifications alter the binding properties and host specificities of the baculovirus to e.g. mammalian cells and help to enhance mammalian cell transduction. Eukaryotic surface display libraries based on the baculovirus system allow selecting for specific binding proteins while providing post translational modifications. Here we describe the different possibilities and strategies to accessibly present foreign proteins and peptides on the surface of insect cells and baculoviruses, as well as the applications thereof, including vector design, cloning strategies and construction and screening of surface expression libraries.
    No preview · Article · Jun 2010 · Current Gene Therapy
Show more

We use cookies to give you the best possible experience on ResearchGate. Read our cookies policy to learn more.