ABSTRACT
Objective:To investigate the role of recombinant phospholipase A2 receptor (PLA2R) tandem dominant epitopes (PLA2RTD) in the removal of anti-PLA2R autoantibodies (anti-PLA2R) from primary membranous nephropathy (PMN).Methods:The recombinant protein PLA2RTD (cysteine-rich domain, C-type lectin like domain 1 and C-type lectin like domain 7) was expressed in bacmid-insect cell expression system. Circular dichroism was used to determine the secondary structure of PLA2RTD. Enzyme-linked immunosorbent assay and immunofluorescence were used to determine the biological activity of PLA2RTD. Epoxy activation method was used to couple the recombinant PLA2RTD and agarose gel CL-6B microspheres for preparing specific immune adsorbent of anti-PLA2R.Results:The study achieved the expression of PLA2RTD in the first time from the bacmid-insect cell system, demonstrating the good immunogenicity and high binding specificity of PLA2RTD. A single in vitro adsorption of PLA2RTD could averagely eliminate 76.66% of anti-PLA2R [(6.66±0.30) RU/ml vs. (28.54±2.10) RU/ml], the changes of IgG, IgA, albumin, β2 microglobulin, interleukin 6, and tumor necrosis factor α were all less than 4% after completion of adsorption, and the second or third repeated use of PLA2RTD could maintain the adsorption efficiency of about 65%. Conclusion:PLA2RTD-based specific immunosorbent can effectively remove anti-PLA2R in plasma, which provides a new way to specifically remove PMN-related autoantibodies.
ABSTRACT
The rapid development of bioeconomy urgently needs the support of biotechnology talents. Establishing an innovative training mode of biotechnology talents can provide support for regional economic development and industrial upgrading. Closely revolved around the concepts of new engineering disciplines development, such as serving the national strategy, docking industry, leading the future development and student-centered, a new economy-oriented training system was developed in School of Bioengineering of Dalian University of Technology. These systems include interdisciplinary curriculum system reconstruction, project-based teaching mode reform, evaluation system implementation and other aspects. The reform and exploration of the first-class biotechnology major under the new economic situation, puts forward the theory of value guidance, deep foundation, strong sense of innovation, technical and non-technical core ability literacy. This reform meets the industry demand for talent diversification, personalization, and dynamic change, helps the merge of industry and education, which provides a way for fostering first-class biotechnology-majored undergraduates.