Baculovirus infection reduced UDP-galactose level in spodoptera frugiperda insect cells
Incomplete glycosylation is always an issue in the expression of recombinant glycoprotein in Baculovirus-Insect Cell Expression System (BEVS). The factors that ensure successful glycosylations are in the presence of sufficient amount of glycosyltransferases, sugar nucleotides as the substance donor...
| Main Authors: | , , |
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| Format: | Conference or Workshop Item |
| Language: | English |
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2005
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| Online Access: | http://eprints.utm.my/5410/1/YapWeiNey2005_BaculovirusInfectionReducedUDP-GalactoseLlevel.pdf |
| _version_ | 1825909733354110976 |
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| author | Yap, Wei Ney Abdul Rahman, Badarulhisam Abdul Aziz, Azila |
| author_facet | Yap, Wei Ney Abdul Rahman, Badarulhisam Abdul Aziz, Azila |
| author_sort | Yap, Wei Ney |
| collection | ePrints |
| description | Incomplete glycosylation is always an issue in the expression of recombinant glycoprotein in Baculovirus-Insect Cell Expression System (BEVS). The factors that ensure successful glycosylations are in the presence of sufficient amount of glycosyltransferases, sugar nucleotides as the substance donor and the recombinant protein as the substrate acceptor. Insect cell lines have been reported to produce small amount of β-1,4galactosyltransferase (β-1,4GaIT) and thus insufficient for effective galactosylation. In our approach, recombinant β-1, 4GalT is being introduced during protein expression by the co-infection with baculovirus carrying, bovine β-1, 4GalT. In this paper, we report our finding on native UDP-Galactose level at normal and upon baculovirus infection in Spodoptera frugiperda (Sf-9) insect cells. We established and monitored native UDP-Galactose content in Sf-9 insect cells using Reverse Phase High Performance Liquid Chromatography. It was found that UDP-Galactose concentration decreased gradually once infected with the recombinant baculovirus. Although UDP-Galactose content was at 0.009 mg/ml prior infection, the level dropped to almost zero upon five days of infection and thus insufficient for effective galactosylation. This interesting finding suggests that the introduction of β-1,4GalT alone is not sufficient for successful galactosylation. |
| first_indexed | 2024-03-05T18:06:34Z |
| format | Conference or Workshop Item |
| id | utm.eprints-5410 |
| institution | Universiti Teknologi Malaysia - ePrints |
| language | English |
| last_indexed | 2024-03-05T18:06:34Z |
| publishDate | 2005 |
| record_format | dspace |
| spelling | utm.eprints-54102010-06-01T15:30:19Z http://eprints.utm.my/5410/ Baculovirus infection reduced UDP-galactose level in spodoptera frugiperda insect cells Yap, Wei Ney Abdul Rahman, Badarulhisam Abdul Aziz, Azila T Technology (General) Incomplete glycosylation is always an issue in the expression of recombinant glycoprotein in Baculovirus-Insect Cell Expression System (BEVS). The factors that ensure successful glycosylations are in the presence of sufficient amount of glycosyltransferases, sugar nucleotides as the substance donor and the recombinant protein as the substrate acceptor. Insect cell lines have been reported to produce small amount of β-1,4galactosyltransferase (β-1,4GaIT) and thus insufficient for effective galactosylation. In our approach, recombinant β-1, 4GalT is being introduced during protein expression by the co-infection with baculovirus carrying, bovine β-1, 4GalT. In this paper, we report our finding on native UDP-Galactose level at normal and upon baculovirus infection in Spodoptera frugiperda (Sf-9) insect cells. We established and monitored native UDP-Galactose content in Sf-9 insect cells using Reverse Phase High Performance Liquid Chromatography. It was found that UDP-Galactose concentration decreased gradually once infected with the recombinant baculovirus. Although UDP-Galactose content was at 0.009 mg/ml prior infection, the level dropped to almost zero upon five days of infection and thus insufficient for effective galactosylation. This interesting finding suggests that the introduction of β-1,4GalT alone is not sufficient for successful galactosylation. 2005 Conference or Workshop Item PeerReviewed application/pdf en http://eprints.utm.my/5410/1/YapWeiNey2005_BaculovirusInfectionReducedUDP-GalactoseLlevel.pdf Yap, Wei Ney and Abdul Rahman, Badarulhisam and Abdul Aziz, Azila (2005) Baculovirus infection reduced UDP-galactose level in spodoptera frugiperda insect cells. In: 15th MSMBB Annual Scientific Meeting, 4-5 April 2005, Kuala Lumpur. |
| spellingShingle | T Technology (General) Yap, Wei Ney Abdul Rahman, Badarulhisam Abdul Aziz, Azila Baculovirus infection reduced UDP-galactose level in spodoptera frugiperda insect cells |
| title | Baculovirus infection reduced UDP-galactose level in spodoptera frugiperda insect cells |
| title_full | Baculovirus infection reduced UDP-galactose level in spodoptera frugiperda insect cells |
| title_fullStr | Baculovirus infection reduced UDP-galactose level in spodoptera frugiperda insect cells |
| title_full_unstemmed | Baculovirus infection reduced UDP-galactose level in spodoptera frugiperda insect cells |
| title_short | Baculovirus infection reduced UDP-galactose level in spodoptera frugiperda insect cells |
| title_sort | baculovirus infection reduced udp galactose level in spodoptera frugiperda insect cells |
| topic | T Technology (General) |
| url | http://eprints.utm.my/5410/1/YapWeiNey2005_BaculovirusInfectionReducedUDP-GalactoseLlevel.pdf |
| work_keys_str_mv | AT yapweiney baculovirusinfectionreducedudpgalactoselevelinspodopterafrugiperdainsectcells AT abdulrahmanbadarulhisam baculovirusinfectionreducedudpgalactoselevelinspodopterafrugiperdainsectcells AT abdulazizazila baculovirusinfectionreducedudpgalactoselevelinspodopterafrugiperdainsectcells |