Which vaccine is produced in insect cells?
The first human vaccine produced in insect cells, CERVARIX, was licensed by the European Medicines Agency (EMA) in 2007 and by U.S. Food and Drug Administration (FDA) in 2009. CERVARIX is a bivalent human papilloma virus vaccine indicated for the prevention of cervical cancers (see Table 1).
Can insects be used for recombinant protein production?
Baculovirus-mediated expression in insect cells has become well-established for the production of recombinant glycoproteins. Its frequent use arises from the relative ease and speed with which a heterologous protein can be expressed on the laboratory scale and the high chance of obtaining a biologically active protein.
Why are insect cells are becoming a popular host cell system for recombinant protein production?
Cell lines have also been developed from insects (Fig. 1.21). They are primarily used to express heterologous proteins for the biotechnology industry. Insect cells are preferred to mammalian cells because they require fewer nutrients for growth and survive in media free of serum.
How do you lyse an insect cell?
Insect cells don’t have a cell wall and therefore lyse rapidly. The most used method is disrupting the cells using a Dounce homogenizer in the presence of a low concentration of detergent (0.1% Triton X-100 or NP-40). Usually 10-20 strokes from the homogenizer are sufficient.
What type of vaccine is Cervarix?
Cervarix is an adjuvanted non-infectious recombinant vaccine prepared from the highly purified virus- like particles (VLPs) of the major capsid L1 protein of oncogenic HPV types 16 and 18. Since the VLPs contain no viral DNA, they cannot infect cells, reproduce or cause disease.
What is Spodoptera frugiperda cell proteins?
Sf9 cells, a clonal isolate of Spodoptera frugiperda Sf21 cells (IPLB-Sf21-AE), are commonly used in insect cell culture for recombinant protein production using baculovirus. They were originally established from ovarian tissue. They can be grown in the absence of serum, and can be cultured attached or in suspension.
What is the advantages of using insect cells for the production of protein?
Insect cells offer high levels of protein expression with posttranslational modification approaching that of mammalian cells, ease of scale-up, and simplified cell growth that can be readily adapted to high-density suspension culture for large-scale expression.
Why is it advantageous to produce proteins in insect cells compared to producing them in bacteria?
Insect Cells The technology has unique biological advantages over bacterial, yeast, or mammalian protein expression systems. A major advantage is the quick turnaround time for the expression of recombinant proteins that show biological activity, antigenicity, and immunogenicity similar to authentic natural proteins.
What are the advantages of protein expression in insect cells?
Insect cells present several comparative advantages to mammalian cells, such as ease of culture, higher tolerance to osmolality and by-product concentration and higher expression levels when infected with a recombinant baculovirus.
How do you express protein in insect cells?
Once you have generated your high-titer viral stocks, you can use Sf9, Sf21, High Five, or Invitrogen Mimic Sf9 cells for protein expression. The need for transforming bacteria and isolating a large bacmid, or cotransfection of a transfer vector and linear baculovirus DNA into insect cells is eliminated.
How do you lyse mammalian cells?
The freeze-thaw method is commonly used to lyse bacterial and mammalian cells. The technique involves freezing a cell suspension in a dry ice/ethanol bath or freezer and then thawing the material at room temperature or 37°C.
How do you lyse mammalian cells for protein purification?