Figure 4 | Scientific Reports

Figure 4

From: Multimerization is required for antigen binding activity of an engineered IgM/IgG chimeric antibody recognizing a skin-related antigen

Figure 4

Results of gel filtration, ELISA and Western blotting analyses of IgM/IgG molecules. (a) Presentation of gel filtration chromatographs for the indicated IgM/IgG hybrid molecules. As a standard, the mixture of recombinant mouse IgG2a (IgG) and original KM48 IgM antibody (IgM) was also analyzed. For ease of comparison, the chromatograph for IgM/IgG mixture was aligned under each of the chromatographs for the other molecules. The elution positions of monomers are indicated. (b) The concentrations of Ig in gel filtration fractions. Concentrations were determined by our novel IgM/IgG sandwich ELISA system for the indicated IgM/IgG molecules. Fractions 7 to 20 were analyzed by ELISA. (c) Non-reducing Western blotting analysis of Ig in fractions. Fractions with sufficient Ig concentration were used. Each non-fractionated sample (O) and hybridoma-derived IgM molecule (IgM) were used as controls. Cropped blots are shown and full length blots are shown in Supplementary Fig. S3.

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