High Five cells

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High Five (BTI-Tn-5B1-4) is an insect cell line that originated from the ovarian cells of the cabbage looper, Trichoplusia ni. [1] It was developed by the Boyce Thompson Institute for Plant Research.

High Five cells have become one of the most commonly used cell lines for recombinant protein expression using baculovirus or transfection, and have been demonstrated to express more recombinant protein than other lepidopteran cell lines, such as Sf9 cells. [2] [3] [4] [5] [6] [7] [8] The High Five cells have been used to produce the VLP-based HPV vaccine Cervarix. [9]

They can be grown in the absence of serum, and can be cultured in a loose attached state or in suspension [10] High Five cells produce abundant microRNAs (miRNAs), small interfering RNAs (siRNAs), and PIWI-interacting RNAs (piRNAs), making it suitable to study all three types of small silencing RNAs. [11]

The High Five cell line, like other cell lines, [12] has been found to host a non-pathogenic adventitious alphanodavirus. Researchers at the Boyce Thompson Institute were able to isolate virus-free sub-clones, named Tnao38 [13] [14] and Tnms42. [15]

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<span class="mw-page-title-main">Protein production</span>

Protein production is the biotechnological process of generating a specific protein. It is typically achieved by the manipulation of gene expression in an organism such that it expresses large amounts of a recombinant gene. This includes the transcription of the recombinant DNA to messenger RNA (mRNA), the translation of mRNA into polypeptide chains, which are ultimately folded into functional proteins and may be targeted to specific subcellular or extracellular locations.

<span class="mw-page-title-main">Chinese hamster ovary cell</span> Cell line

Chinese hamster ovary (CHO) cells are an epithelial cell line derived from the ovary of the Chinese hamster, often used in biological and medical research and commercially in the production of recombinant therapeutic proteins. They have found wide use in studies of genetics, toxicity screening, nutrition and gene expression, particularly to express recombinant proteins. CHO cells are the most commonly used mammalian hosts for industrial production of recombinant protein therapeutics.

<span class="mw-page-title-main">Expression vector</span> Virus or plasmid designed for gene expression in cells

An expression vector, otherwise known as an expression construct, is usually a plasmid or virus designed for gene expression in cells. The vector is used to introduce a specific gene into a target cell, and can commandeer the cell's mechanism for protein synthesis to produce the protein encoded by the gene. Expression vectors are the basic tools in biotechnology for the production of proteins.

Human embryonic kidney 293 cells, also often referred to as HEK 293, HEK-293, 293 cells, or less precisely as HEK cells, are a specific immortalised cell line derived from a spontaneously miscarried or aborted fetus or human embryonic kidney cells grown in tissue culture taken from a female fetus in 1973.

<span class="mw-page-title-main">Recombinant DNA</span> DNA molecules formed by human agency at a molecular level generating novel DNA sequences

Recombinant DNA (rDNA) molecules are DNA molecules formed by laboratory methods of genetic recombination that bring together genetic material from multiple sources, creating sequences that would not otherwise be found in the genome.

Virus-like particles (VLPs) are molecules that closely resemble viruses, but are non-infectious because they contain no viral genetic material. They can be naturally occurring or synthesized through the individual expression of viral structural proteins, which can then self assemble into the virus-like structure. Combinations of structural capsid proteins from different viruses can be used to create recombinant VLPs. Both in-vivo assembly and in-vitro assembly have been successfully shown to form virus-like particles. VLPs derived from the Hepatitis B virus (HBV) and composed of the small HBV derived surface antigen (HBsAg) were described in 1968 from patient sera. VLPs have been produced from components of a wide variety of virus families including Parvoviridae, Retroviridae, Flaviviridae, Paramyxoviridae and bacteriophages. VLPs can be produced in multiple cell culture systems including bacteria, mammalian cell lines, insect cell lines, yeast and plant cells.

<i>Baculoviridae</i> Family of viruses

Baculoviridae is a family of viruses. Arthropods, among the most studied being Lepidoptera, Hymenoptera and Diptera, serve as natural hosts. Currently, 85 species are placed in this family, assigned to four genera.

<span class="mw-page-title-main">Vero cell</span>

Vero cells are a lineage of cells used in cell cultures. The 'Vero' lineage was isolated from kidney epithelial cells extracted from an African green monkey. The lineage was developed on 27 March 1962, by Yasumura and Kawakita at the Chiba University in Chiba, Japan. The original cell line was named Vero after an abbreviation of verdareno, which means 'green kidney' in Esperanto, while vero itself means 'truth' in Esperanto.

<span class="mw-page-title-main">Sf21</span> Cell line

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<span class="mw-page-title-main">Agroinfiltration</span>

Agroinfiltration is a method used in plant biology and especially lately in plant biotechnology to induce transient expression of genes in a plant, or isolated leaves from a plant, or even in cultures of plant cells, in order to produce a desired protein. In the method, a suspension of Agrobacterium tumefaciens is introduced into a plant leaf by direct injection or by vacuum infiltration, or brought into association with plant cells immobilised on a porous support, whereafter the bacteria transfer the desired gene into the plant cells via transfer of T-DNA. The main benefit of agroinfiltration when compared to the more traditional plant transformation is speed and convenience, although yields of the recombinant protein are generally also higher and more consistent.

<span class="mw-page-title-main">Influenza</span> Infectious disease, often just "the flu"

Influenza, commonly known as "the flu", is an infectious disease caused by influenza viruses. Symptoms range from mild to severe and often include fever, runny nose, sore throat, muscle pain, headache, coughing, and fatigue. These symptoms begin from one to four days after exposure to the virus and last for about 2–8 days. Diarrhea and vomiting can occur, particularly in children. Influenza may progress to pneumonia, which can be caused by the virus or by a subsequent bacterial infection. Other complications of infection include acute respiratory distress syndrome, meningitis, encephalitis, and worsening of pre-existing health problems such as asthma and cardiovascular disease.

A subunit vaccine is a vaccine that contains purified parts of the pathogen that are antigenic, or necessary to elicit a protective immune response. Subunit vaccine can be made from dissembled viral particles in cell culture or recombinant DNA expression, in which case it is a recombinant subunit vaccine.

<span class="mw-page-title-main">BacMam</span>

Baculovirus gene transfer into Mammalian cells, known from scientific research articles as BacMam, is the use of baculovirus to deliver genes to mammalian cells. Baculoviruses are insect viruses that can be modified to express proteins in mammalian cells. The unmodified baculovirus is able to enter those cells; however, its genes are not expressed unless a mammalian recognizable promoter is incorporated upstream of a gene of interest. Both unmodified baculovirus and its modified counterpart are unable to replicate in humans and are thus non-infectious.

<span class="mw-page-title-main">Genetically modified virus</span> Species of virus

A genetically modified virus is a virus that has been altered or generated using biotechnology methods, and remains capable of infection. Genetic modification involves the directed insertion, deletion, artificial synthesis or change of nucleotide bases in viral genomes. Genetically modified viruses are mostly generated by the insertion of foreign genes intro viral genomes for the purposes of biomedical, agricultural, bio-control, or technological objectives. The terms genetically modified virus and genetically engineered virus are used synonymously.

Heterologous expression refers to the expression of a gene or part of a gene in a host organism that does not naturally have the gene or gene fragment in question. Insertion of the gene in the heterologous host is performed by recombinant DNA technology. The purpose of heterologous expression is often to determine the effects of mutations and differential interactions on protein function. It provides an easy path to efficiently express and experiment with combinations of genes and mutants that do not naturally occur.

<span class="mw-page-title-main">Sf9 (cells)</span> Insect cell line

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.

The use of insect cell lines as production hosts is an emerging technology for the production of bio pharmaceuticals. There are currently more than 100 insect cell lines available for recombinant protein production with lines derived from Bombyx mori, Mamestra brassicae, Spodoptera frugiperda, Trichoplusia ni, and Drosophila melanogaster being of particular interest. Insects cell lines are commonly used in place of prokaryotic ones because post-translational modifications of proteins are possible in insect cells whereas this mechanism is not present in prokaryotic systems. The Sf9 cell line is one of the most commonly used lines in insect cell culture.

Flock House virus (FHV) is in the Alphanodavirus genus of the Nodaviridae family of viruses. Flock House virus was isolated from a grass grub at the Flock House research station in Bulls, New Zealand. FHV is an extensively studied virus and is considered a model system for the study of other non-enveloped RNA viruses owing to its small size and genetic tractability, particularly to study the role of the transiently exposed hydrophobic gamma peptide and the metastability of the viral capsid. FHV can be engineered in insect cell culture allowing for the tailored production of native or mutant authentic virions or virus-like-particles. FHV is a platform for nanotechnology and nanomedicine, for example, for epitope display and vaccine development. Viral entry into host cells occurs via receptor-mediated endocytosis. Receptor binding initiates a sequence of events during which the virus exploits the host environment in order to deliver the viral cargo in to the host cytosol. Receptor binding prompts the meta-stability of the capsid–proteins, the coordinated rearrangements of which are crucial for subsequent steps in the infection pathway. In addition, the transient exposure of a covalently-independent hydrophobic γ-peptide is responsible for breaching cellular membranes and is thus essential for the viral entry of FHV into host cells.

Transient expression, more frequently referred to "transient gene expression", is the temporary expression of genes that are expressed for a short time after nucleic acid, most frequently plasmid DNA encoding an expression cassette, has been introduced into eukaryotic cells with a chemical delivery agent like calcium phosphate (CaPi) or polyethyleneimine (PEI). However, unlike "stable expression," the foreign DNA does not fuse with the host cell DNA, resulting in the inevitable loss of the vector after several cell replication cycles. The majority of transient gene expressions are done with cultivated animal cells. The technique is also used in plant cells; however, the transfer of nucleic acids into these cells requires different methods than those with animal cells. In both plants and animals, transient expression should result in a time-limited use of transferred nucleic acids, since any long-term expression would be called "stable expression."

Max Duane Summers is an American molecular biologist and inventor, known for his work on the Baculovirus Expression Vector System (BEVS).

References

  1. ↑ Wickham TJ, Davis T, Granados RR, Shuler ML, Wood HA (1992). "Screening of insect cell lines for the production of recombinant proteins and infectious virus in the baculovirus expression system". Biotechnol. Prog. 8 (5): 391–396. doi:10.1021/bp00017a003. PMID   1369220. S2CID   41111655.
  2. ↑ Davis TR, Trotter KM, Granados RR, Wood HA (1992). "Baculovirus expression of alkaline phosphatase as a reporter gene for evaluation of production, glycosylation and secretion". Bio/Technology. 10 (10): 1148–1150. doi:10.1038/nbt1092-1148. PMID   1368794. S2CID   5978915.
  3. ↑ Wickham TJ, Nemerow GR (1993). "Optimization of growth methods and recombinant protein production in BTI-Tn-5B1-4 insect cells using the baculovirus expression system". Biotechnol. Prog. 9 (1): 25–30. doi:10.1021/bp00019a004. PMID   7764044. S2CID   21157206.
  4. ↑ Granados RR, Guoxun L, Derksen AC, McKenna KA (1994). "A New Insect Cell Line from Trichoplusia ni (BTI-Tn-5B1-4) Susceptible to Trichoplusia ni Single Enveloped Nuclear Polyhedrosis". Virus. J. Invertebr. Pathol. 64: 260–266. doi: 10.1016/s0022-2011(94)90400-6 .
  5. ↑ Lai, Chia-Chun; Cheng, Yu-Chieh; Chen, Pin-Wen; Lin, Ting-Hui; Tzeng, Tsai-Teng; Lu, Chia-Chun; Lee, Min-Shi; Hu, Alan Yung-Chih (December 2019). "Process development for pandemic influenza VLP vaccine production using a baculovirus expression system". Journal of Biological Engineering. 13 (1): 78. doi: 10.1186/s13036-019-0206-z . ISSN   1754-1611. PMC   6813129 . PMID   31666806.
  6. ↑ Krammer, Florian; Schinko, Theresa; Palmberger, Dieter; Tauer, Christopher; Messner, Paul; Grabherr, Reingard (July 2010). "Trichoplusia ni cells (High FiveTM) are highly efficient for the production of influenza A virus-like particles: a comparison of two insect cell lines as production platforms for influenza vaccines". Molecular Biotechnology. 45 (3): 226–234. doi:10.1007/s12033-010-9268-3. ISSN   1073-6085. PMC   4388404 . PMID   20300881.
  7. ↑ Krammer, Florian; Nakowitsch, Sabine; Messner, Paul; Palmberger, Dieter; Ferko, Boris; Grabherr, Reingard (January 2010). "Swine-origin pandemic H1N1 influenza virus-like particles produced in insect cells induce hemagglutination inhibiting antibodies in BALB/c mice". Biotechnology Journal. 5 (1): 17–23. doi:10.1002/biot.200900267. PMC   4388400 . PMID   20041443.
  8. ↑ Wilde, Monika; Klausberger, Miriam; Palmberger, Dieter; Ernst, Wolfgang; Grabherr, Reingard (April 2014). "Tnao38, high five and Sf9—evaluation of host–virus interactions in three different insect cell lines: baculovirus production and recombinant protein expression". Biotechnology Letters. 36 (4): 743–749. doi:10.1007/s10529-013-1429-6. ISSN   0141-5492. PMC   3955137 . PMID   24375231.
  9. ↑ Schiller, John T.; Castellsagué, Xavier; Villa, Luisa L.; Hildesheim, Allan (August 2008). "An update of prophylactic human papillomavirus L1 virus-like particle vaccine clinical trial results". Vaccine. 26 (Suppl 10): K53–K61. doi:10.1016/j.vaccine.2008.06.002. PMC   2631230 . PMID   18847557.
  10. ↑ Invitrogen (31 March 2015). "Cell Lines". Growth and Maintenance of Insect cell lines (PDF). A.0. Thermo Fisher Scientific. Retrieved 2015-11-09.
  11. ↑ Fu, Yu; Yang, Yujing; Zhang, Han; Farley, Gwen; Wang, Junling; Quarles, Kaycee A; Weng, Zhiping; Zamore, Phillip D (2018-01-29). "The genome of the Hi5 germ cell line from Trichoplusia ni, an agricultural pest and novel model for small RNA biology". eLife. 7. doi: 10.7554/eLife.31628 . ISSN   2050-084X. PMC   5844692 . PMID   29376823.
  12. ↑ Geisler, Christoph; Jarvis, Donald L. (April 2018). "Adventitious viruses in insect cell lines used for recombinant protein expression". Protein Expression and Purification. 144: 25–32. doi:10.1016/j.pep.2017.11.002. PMC   5826799 . PMID   29133148.
  13. ↑ Hashimoto, Yoshifumi; Zhang, Sheng; Blissard, Gary W (December 2010). "Ao38, a new cell line from eggs of the black witch moth, Ascalapha odorata (Lepidoptera: Noctuidae), is permissive for AcMNPV infection and produces high levels of recombinant proteins". BMC Biotechnology. 10 (1): 50. doi: 10.1186/1472-6750-10-50 . ISSN   1472-6750. PMC   2906426 . PMID   20602790.
  14. ↑ Hashimoto, Yoshi; Zhang, Sheng; Zhang, Shiying; Chen, Yun-Ru; Blissard, Gary W (December 2012). "Erratum to: BTI-Tnao38, a new cell line derived from Trichoplusia ni, is permissive for AcMNPV infection and produces high levels of recombinant proteins". BMC Biotechnology. 12 (1): 12. doi: 10.1186/1472-6750-12-12 . ISSN   1472-6750. PMC   3376038 . PMID   22531032.
  15. ↑ Koczka, Krisztina; Peters, Philipp; Ernst, Wolfgang; Himmelbauer, Heinz; Nika, Lisa; Grabherr, Reingard (March 2018). "Comparative transcriptome analysis of a Trichoplusia ni cell line reveals distinct host responses to intracellular and secreted protein products expressed by recombinant baculoviruses". Journal of Biotechnology. 270: 61–69. doi:10.1016/j.jbiotec.2018.02.001. PMID   29432775.
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