Screening of the Sf900-II SFM insect cell culture medium and recombinant baculovirus for the expression of recombinant human transferrin

Optimized medium composition and high purity recombinant baculovirus stock have historically contributed to high yield recombinant protein expressed in the insect cells baculovirus expression system. In achieving this, screening experiments were conducted to identify the recombinant baculovirus and...

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Main Authors: M. Ongkudon, Clarence, Abdul Rahman, Badarulhisam, Abdul Aziz, Azila
Format: Conference or Workshop Item
Language:English
Published: 2005
Subjects:
Online Access:http://eprints.utm.my/5415/1/ClarenceM.Ongkudon2005_ScreeningOfTheSf900-IISFMInsectCell.pdf
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author M. Ongkudon, Clarence
Abdul Rahman, Badarulhisam
Abdul Aziz, Azila
author_facet M. Ongkudon, Clarence
Abdul Rahman, Badarulhisam
Abdul Aziz, Azila
author_sort M. Ongkudon, Clarence
collection ePrints
description Optimized medium composition and high purity recombinant baculovirus stock have historically contributed to high yield recombinant protein expressed in the insect cells baculovirus expression system. In achieving this, screening experiments were conducted to identify the recombinant baculovirus and medium components in Sf900-II SFM that gave significant production of recombinant human transferrin (rhTf). For medium screening, a Plackett-Burman screening experiments were conducted. For recombinant baculovirus screenings, an end point dilution method based on Poisson distribution was used. RhTf was analyzed using SDS-PAGE and ELISA while the software Statistica (Statsoft, v. 5.0) was used to analyze the results obtained. The results show that glucose and glutamine had the most effect with more than 95% significance, while the recombinant baculovirus was identified at 98.7% purity. The two screening methods were very useful to rapidly screen recombinant baculovirus and culture medium. At the same time, the recombinant protein yield was increased to ~20 ug/ml.
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spelling utm.eprints-54152017-08-30T04:31:29Z http://eprints.utm.my/5415/ Screening of the Sf900-II SFM insect cell culture medium and recombinant baculovirus for the expression of recombinant human transferrin M. Ongkudon, Clarence Abdul Rahman, Badarulhisam Abdul Aziz, Azila T Technology (General) Optimized medium composition and high purity recombinant baculovirus stock have historically contributed to high yield recombinant protein expressed in the insect cells baculovirus expression system. In achieving this, screening experiments were conducted to identify the recombinant baculovirus and medium components in Sf900-II SFM that gave significant production of recombinant human transferrin (rhTf). For medium screening, a Plackett-Burman screening experiments were conducted. For recombinant baculovirus screenings, an end point dilution method based on Poisson distribution was used. RhTf was analyzed using SDS-PAGE and ELISA while the software Statistica (Statsoft, v. 5.0) was used to analyze the results obtained. The results show that glucose and glutamine had the most effect with more than 95% significance, while the recombinant baculovirus was identified at 98.7% purity. The two screening methods were very useful to rapidly screen recombinant baculovirus and culture medium. At the same time, the recombinant protein yield was increased to ~20 ug/ml. 2005 Conference or Workshop Item PeerReviewed application/pdf en http://eprints.utm.my/5415/1/ClarenceM.Ongkudon2005_ScreeningOfTheSf900-IISFMInsectCell.pdf M. Ongkudon, Clarence and Abdul Rahman, Badarulhisam and Abdul Aziz, Azila (2005) Screening of the Sf900-II SFM insect cell culture medium and recombinant baculovirus for the expression of recombinant human transferrin. In: 15th MSMBB Annual Scientific Meeting, 4-5 April 2005, Kuala Lumpur.
spellingShingle T Technology (General)
M. Ongkudon, Clarence
Abdul Rahman, Badarulhisam
Abdul Aziz, Azila
Screening of the Sf900-II SFM insect cell culture medium and recombinant baculovirus for the expression of recombinant human transferrin
title Screening of the Sf900-II SFM insect cell culture medium and recombinant baculovirus for the expression of recombinant human transferrin
title_full Screening of the Sf900-II SFM insect cell culture medium and recombinant baculovirus for the expression of recombinant human transferrin
title_fullStr Screening of the Sf900-II SFM insect cell culture medium and recombinant baculovirus for the expression of recombinant human transferrin
title_full_unstemmed Screening of the Sf900-II SFM insect cell culture medium and recombinant baculovirus for the expression of recombinant human transferrin
title_short Screening of the Sf900-II SFM insect cell culture medium and recombinant baculovirus for the expression of recombinant human transferrin
title_sort screening of the sf900 ii sfm insect cell culture medium and recombinant baculovirus for the expression of recombinant human transferrin
topic T Technology (General)
url http://eprints.utm.my/5415/1/ClarenceM.Ongkudon2005_ScreeningOfTheSf900-IISFMInsectCell.pdf
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