Stabilised proteins for immunising against staphylococcus aureus
一种蛋白、免疫原性的技术,应用在抗细菌药、含有效成分的医用配制品、过敏性疾病等方向,能够解决疫苗不一致性影响、抗原不稳定、疫苗生产复杂等问题
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[0200] heat denaturation test
[0201] The Sta006Cys(+) antigen used in the experiments described below is represented as SEQ ID NO:3 and the Sta006Cys(-) antigen is represented as SEQ ID NO:6. Both antigens are recombinant proteins purified from E. coli.
[0202] The thermostability of Sta006 cysteine-containing Cys(+) antigen and Sta006 cysteine-deficient Cys(-) antigen was compared by differential scanning fluorescence assay (DSF). Antigen-containing samples (10 μΜ in PBS) were heated under controlled conditions in a Strategen Mx3000p real-time PCR instrument at a temperature ramp rate of 1 °C / min. The dye SyproOrange 5x was used and the change in fluorescence was monitored. The tests were carried out in the temperature range of 10-100°C.
[0203] figure 1 Melting curves for the tested antigens are reported. It shows that the peak of Cys(-) antigen is slightly shifted to the top and left compared to Cys(+) antigen. Melting temperature (Tm) was determined by fitting the ...
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