Skip to main content
Fig. 4 | Journal of Translational Medicine

Fig. 4

From: Identification of anti-SF3B1 autoantibody as a diagnostic marker in patients with hepatocellular carcinoma

Fig. 4

XC24p11-streptavidin mimicked effectively the antigenic structure on SF3B1 for the binding of XC24 autoantibody. a Construction of XC24p11 cyclic peptide-fused streptavidin (XC24p11-STA) expression vector. Dsb A, a bacterial thiol disulfide oxidoreductase, was co-expressed to enhance disulfide bond formation of the XC24p11 cyclic epitope; b SDS-PAGE and western blot analysis of XC24p11-STA (5 or 1 μg/each lane). To confirm the monomeric form of XC24p11-STA, purified XC24p11-STA was mixed with reducing SDS-PAGE sample buffer, boiled, and analyzed by 10% SDS-PAGE and western blotting. Tetrameric XC24p11-STA was also confirmed by the analysis of non-reduced and non-boiled antigen in SDS-PAGE sample buffer on 10% SDS-PAGE and western blotting (CBB, Coomassie Brilliant Blue staining; α-STA, anti-streptavidin antibody staining; α-HIS, anti-His6 antibody staining; XC24, XC24 autoantibody staining). K94p1-STA, another autoantigenic epitope-fused STA (K94p1 epitope, -CISPDAHSC-, previously reported [31]) and epitope-free STA (Eph-STA) were also analyzed as controls; c XC24p11-STA ELISA on MaxiSorp- or biotin-coated plates. The conditions of ELISA are described in detail in Materials and Methods

Back to article page