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KMID : 0380020090240010080
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Korean Journal of Biotechnology and Bioengineering
2009 Volume.24 No. 1 p.80 ~ p.88
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Purification of Recombinant CTP-Conjugated Human prostatic acid phosphatase for activation of Dendritic Cell
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Yi Ki-Wan
Ryu Kang
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Abstract
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Human prostatic acid phosphatase (PAP), with comprehensive homology to glandular kallikrein, are representative serum biomarkers of prostate cancer. Dendritic cell (DC), which is the potent antigen-presenting cells(APC) in the immune system, can induce strong T cell responses against viruses, microbial pathogens, and tumors. Therefore, the immunization using DC loaded with tumor-associated antigens is a powerful method for inducing anti-tumor immunity. The CTP (Cytoplasmic Transduction Peptide) technology developed by Creagene which can transport attached bio-polymers like nucleic acids or proteins into the cell with high permeation efficiency. As the active forms of PAP can mediate apoptotic processing, we used multimer forms of PAP as an inactive form for antigen pulsing of DCs. In this study, multimeric forms of CTP-rhPAP was obtained according to the advanced purification process and subsequently confirmed by gel filtration chromatography, western blot and Dynamic Light Scattering. Therefore, CTP-conjugated PA multimers transduced into the cytoplasm were efficiently presented on the cell surface without any harm effect on cells via MHC class I molecules and result in induction of a large number of effector cell.
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KEYWORD
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Human prostatic acid phosphatase, Dendritic cell, antigen, Cytoplasmic Transduction Peptide, multimer, nanoparticle
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