Comparative proteomic analysis for hCTLA4Ig production in transgenic rice suspension cultures using two-dimensional difference gel electrophoresis |
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Authors: | Ji-Suk Cho Hye Won Lee Song-Jae Lee Dong-Il Kim |
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Institution: | 1.Department of Biological Engineering,Inha University,Incheon,Korea;2.GE Healthcare Bio-Sciences Korea,Seoul,Korea;3.Boryung Pharmaceutical Co. Ltd.,Ansan,Korea |
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Abstract: | The new technology, two-dimensional difference gel electrophoresis (2D DIGE), uses fluorescent dyes to simplify the process
of detecting and matching proteins between multiple gels by allowing for the separation of up to three separate protein samples
within the same gel. In this study, recombinant human cytotoxic T lymphocyte-associated antigen 4-immunoglobulin (hCTLA4lg)
was produced in transgenic rice suspension cell cultures and the intracellular proteins were analyzed by 2D DIGE. The highest
level of hCTLA4Ig (25.4 mg/L) was obtained five days after induction. The intracellular proteins expressed at both the growth
and induction culture stages were separated and analyzed using DeCyder software. At least 2,218 spots were detected with two-fold
thresholds and 95% confidence. We found that 29 spots increased and 20 spots decreased in their intensities during the production
of recombinant hCTLA4Ig. In addition, the 2D Western blot of hCTLA4Ig revealed that this fusion protein was expressed in a
variety of isoforms. |
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Keywords: | |
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