Expression of GA733-Fc Fusion Protein as a Vaccine Candidate for Colorectal Cancer in Transgenic Plants

The tumor-associated antigen GA733 is a cell-surface glycoprotein highly expressed in colorectal carcinomas. In this study, 3 recombinant genes were constructed as follows: GA733 tagged to the ER retention sequence KDEL (GA733K), GA733 fused to the immunoglobulin Fc fragment (GA733-Fc), and GA733-Fc fused to the ER retention sequence (GA733-FcK). Agrobacterium-mediated transformation was used to generate transgenic plants expressing recombinant genes. The presence of transgenes was confirmed by genomic PCR. Western blot, confocal immunofluorescence, and sandwich ELISA showed the expression of recombinant proteins. The stability, flexibility, and bioactivity of recombinant proteins were analyzed and demonstrated through N-glycosylation analysis, animal trials, and sera ELISA. Our results suggest that the KDEL retained proteins in ER with oligomannose glycan structure and enhanced protein accumulation level. The sera of mice immunized with GA733-FcK purified from plants contained immunoglobulins which were at least as efficient as the mammalian-derived GA733-Fc at recognizing human colorectal cancer cell lines. Thus, a plant system can be used to express the KDEL fusion protein with oligomannose glycosylation, and this protein induces an immune response which is comparable to non-KDEL-tagged, mammalian-derived proteins

Corrosion behavior and cellular automata simulation of carbon steel in salt-spray environment

Abstract Scanning electron microscope (SEM) and X-ray diffraction (XRD) were used to discuss the corrosion loss and morphology of the pit and rust layer of carbon steel. It was found that the corrosion process is largely influenced by the cyclic shedding of surface corrosion products, in addition to being controlled by the mechanism of oxide film shedding and pit evolution. A corrosion mechanism (the mechanism of rust layer shedding) is proposed. As a result, in this paper, the corrosion process of the test steel is simulated by the cellular automata. It was set up that the mechanism of oxide film shedding, the mechanism of pit evolution, and the mechanism of rust layer shedding in Cellular Automata Simulation. The optimal time ratio and simulation parameters were found, and a predictable cellular automata corrosion simulation model was built, providing a solution for carbon steel’s service life prediction

Chemical Probes Reveal Sirt2’s New Function as a Robust “Eraser” of Lysine Lipoylation

10.1021/jacs.9b06913J. Am. Chem. Soc.1414618428–1843