Expression of a recombinant chimeric protein of human colorectal cancer antigen GA733-2 and Fc fragment of antibody using a replicating vector based on Beet curly top virus in infiltrated Nicotiana benthamiana leaves

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Title
Expression of a recombinant chimeric protein of human colorectal cancer antigen GA733-2 and Fc fragment of antibody using a replicating vector based on Beet curly top virus in infiltrated Nicotiana benthamiana leaves
Author(s)
K I Kim; H Y Chung; K H Yoo; J H Park; H H Lee; Y J Seok; K S Ko; H S Kang; Kyung Jin Lee; Doo-Byoung Oh; Y H Joung; I S Chung
Bibliographic Citation
Plant Biotechnology Reports, vol. 6, no. 3, pp. 233-242
Publication Year
2012
Abstract
We describe expression and characterization of recombinant human colorectal cancer antigen GA733-2 fused to Fc fragment of antibody (GA733-2-Fc) using a replicating vector based on Beet curly top virus in infiltrated Nicotiana benthamiana leaves. Recombinant GA733-2-Fc/KDEL with a molecular mass of ~68 kDa was transiently expressed. The level of expression of GA733-2-Fc with ER retention signal KDEL (GA733-2-Fc/KDEL) in the expression vector system was 0. 96% of total soluble proteins. Recombinant GA733-2-Fc/KDEL was purified using an affinity chromatography. Mice immunized with recombinant GA733-2-Fc/KDEL mounted a strong GA733-2-Fc/KDEL-specific serum antibody response. Vaccination of plant-derived recombinant GA733-2-Fc/KDEL regressed tumor volumes in BALB/c mice. The population of activated-T and NK-T cells increased notably in lymph node, spleen, and tumor-infiltrating lymphocytes derived from the tumor-regressed mice. Taken together, recombinant GA733-2-Fc/KDEL expressed in plants can be used as an effective experimental immunogen for research in cancer vaccine development.
Keyword
BCTV (Beet curly top virus)Colorectal cancerGA733-2Nicotiana benthamianaVaccination
ISSN
1863-5466
Publisher
Springer
DOI
http://dx.doi.org/10.1007/s11816-012-0218-3
Type
Article
Appears in Collections:
Aging Convergence Research Center > 1. Journal Articles
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