Fig. 4: MFHR13 structure and sequence characterization.
From: A synthetic protein as efficient multitarget regulator against complement over-activation
a Coomassie staining of purified MFHR1I62 and MFHR13 via Ni-affinity and anion exchange chromatography. Samples were separated on SDS-PAGE under reducing conditions. b Semi-quantitative western blot of MFHR1V62, MFHR1I62, and MFHR13 under reducing conditions and detected with anti-His-tag antibodies. A calibration curve was obtained with MFHR1V62 and densities of the western blot signal were compared with the concentrations obtained by ELISA (included under each lane). One representative experiment is shown. c MFHR13 sequence. FHR11-2 are shown in green, FH1-4, FH13, FH19-20 are shown in blue, gray and light red, respectively, and the linkers are shown in light yellow. The glycosylation sites are highlighted in red and the deamidated site in italics. The peptides identified by MS are shown in bold. d Relative quantification of glycopeptides based on MS. MFHR13 was produced in stirred tank bioreactor (5 L) and extracted at day 8. NIS: glycosylation site in the FHR12 in MFHR13. NCS: glycosylation site in the FH13 in MFHR13. Mean values from two technical replicates with standard deviation are shown.
