General Info 11.45.23 PM

General Info 11.45.23 PM - 1 Insect Cell Culture- General...

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1 Insect Cell Culture- General Information The utilization of insect cell culture for heterologous protein expression has steadily increased over the last several decades. It has become a common expression system for both basic research and large-scale commercial applications. Currently, some of the common uses of insect cell expression include the research driven gene expression for protein crystallography-small molecule interaction, and FDA regulated GMP protein production. These regulated applications can include subunit vaccine production as well as the in vitro diagnostic and in vivo therapeutic markets. A key factor to the popularity of insect cell expression is the ability of insect cells to produce relatively large quantities of posttranslationally modified eukaryotic proteins in a relatively short period of time. Most insect cell-produced proteins have been expressed by employing the Baculovirus Expression Vector System (BEVS); however, other technologies that make stable transfected insect cells are gaining in popularity. Many types of viruses infect insects, with the most common belonging to the family Baculoviridae. The most popular invertebrate expression vector system is based on the Autographa Californica nuclear polyhedrosis virus ( AcNPV ), an insect baculovirus isolated from the Alfalfa looper that replicates in the nucleus of over 30 lepidopteran insect cell lines. The baculovirus expression vector system has been used to express genes derived from viruses, fungi, bacteria, plants, and animals. In this system, foreign genes placed under the control of the strong polyhedrin promoter of the AcNPV are usually expressed at high levels in cultured lepidopteran insect cells. Baculovirus constitute one of the largest known groups of viruses, and they are capable of infecting over 500 species of insects, and more recently, these viruses have shown the ability to make ideal vectors for a variety of mammalian cell lines. The most widely used lepidopteran cells for BEVS are the Sf9 and Sf21 cell lines isolated from ovarian tissue of the fall army worm, Spodoptera frugiperda , and the High Five cell line, designated BTI- Tn-5B1-4, originally established from the Trichoplusia ni embryonic tissue. Sf9 cells are a sub-clone of the Sf21 cells and were selected for their faster growth rate and higher cell densities than the Sf21 cells. Spodoptera frugiperda cells, either Sf9 or Sf21 are preferred for virus expansion. Sf21 cells can compare favorably, in terms of heterologous protein expression, to both High Fives and the Sf9 cell lines in certain situations. The BEVS technology, developed by Max Summers, Gale Smith and colleagues at Texas A&M University, 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. Also, the vectors are not dependent
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This note was uploaded on 12/16/2011 for the course BIOLOGY 101 taught by Professor Mr.wallace during the Fall '11 term at Montgomery College.

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General Info 11.45.23 PM - 1 Insect Cell Culture- General...

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