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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.
- Authors
Kim, Kyung; Chung, Ha; Yoo, Ki; Park, Jong-Hwa; Lee, Hyun; Soek, Yeon; Ko, Ki; Kang, Hyung; Lee, Kyung; Oh, Doo-Byoung; Joung, Young; Chung, In
- 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.
- Subjects
NICOTIANA benthamiana; BEET curly top virus; CHIMERIC proteins; AFFINITY chromatography; CANCER vaccines; LABORATORY mice
- Publication
Plant Biotechnology Reports, 2012, Vol 6, Issue 3, p233
- ISSN
1863-5466
- Publication type
Article
- DOI
10.1007/s11816-012-0218-3