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KMID : 0380020160310010033
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Korean Journal of Biotechnology and Bioengineering
2016 Volume.31 No. 1 p.33 ~ p.39
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Live Cell Detection of Monoclonal Antibody Light and Heavy Chain mRNAs using Molecular Beacons
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Jeong Seung-A
Rhee Won-Jong
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Abstract
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Developing the method for the selection of animal cell line producing therapeutic monoclonal antibody (mAb) is invaluable as its market is rapidly growing. Although the quality of produced mAb is as important as quantity, however there is no method developed for the selective screening of cell lines on the basis of both quantity and quality. From recent reports, the ratio of light and heavy chain mRNAs of mAb in the cell is a key parameter for the indication of product quality. Therefore, it is obvious that developing the novel method that can detect both light and heavy chain mRNAs in single live cell will provide unprecedented opportunities in bio-industry. Here, we have constructed oligonucleotide probes, molecular beacons for the detection of light or heavy chain mRNAs, respectively, in the live cells producing mAbs. Both beacons showed increased fluorescent intensity after transient transfection of plasmid expressing mAbs analyzed by fluorometer. Flow cytometric analysis clearly demonstrated that both molecular beacons can simultaneously detect the expression of light and heavy chain mRNAs of mAb in the same cell. The technique described in the thesis provides the new direction and concept for developing the method for the smart selection of cell lines producing recombinant proteins including therapeutic mAbs.
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KEYWORD
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Monoclonal antibody, Cell line, Anti-apoptosis, mRNA, Molecular beacon, Recombinant protein
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