The production of recombinant human CFH protein involves several key steps. First, the CFH gene is cloned into an expression vector, which is then introduced into a suitable expression system, such as yeast, in vivo biotinylation in E.coli, baculovirus, and mammalian cells, for protein expression. Once expressed, the protein undergoes purification using affinity chromatography to isolate the target protein. After purification, SDS-PAGE is used to assess the purity of the recombinant CFH protein, exceeding 85%.
Human CFH is a crucial regulatory protein within the complement system, primarily regulating the alternative pathway of complement activation. CFH plays a pivotal role in maintaining immune homeostasis by preventing excessive complement activation, which can lead to tissue damage and inflammation [1][2]. It achieves this by acting as a cofactor for complement factor I, facilitating the cleavage of C3b into inactive forms, thereby inhibiting the formation of the C3 convertase [3][4]. This regulatory function is vital in protecting host tissues from complement-mediated damage, particularly in age-related diseases.
References:
[1] C. Schmidt, A. Herbert, H. Hocking, D. Uhrı́n, & P. Barlow。 Translational mini-review series on complement factor h: structural and functional correlations for factor h, Clinical & Experimental Immunology, vol. 151, no. 1, p. 14-24, 2007. https://doi.org/10.1111/j.1365-2249.2007.03553.x
[2] F. Vaziri-Sani, L. Holmberg, et al. Phenotypic expression of factor h mutations in patients with atypical hemolytic uremic syndrome, Kidney International, vol. 69, no. 6, p. 981-988, 2006. https://doi.org/10.1038/sj.ki.5000155
[3] A. Mühlig, L. Keir, et al. Podocytes produce and secrete functional complement c3 and complement factor h, Frontiers in Immunology, vol. 11, 2020. https://doi.org/10.3389/fimmu.2020.01833
[4] C. Paun, Y. Lechanteur, et al. A novel complotype combination associates with age-related macular degeneration and high complement activation levels in vivo, Scientific Reports, vol. 6, no. 1, 2016. https://doi.org/10.1038/srep26568
