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. 2021 Apr 4;14(4):330. doi: 10.3390/ph14040330

Figure 1.

Figure 1

Characterization of autoantibodies in mouse and human sera. (A) Determination of isotypes. Antibodies were affinity-purified from pooled sera over human ApoB100 and analyzed with paper strip isotyping kits. Blue arrows indicate positive reactions. C; control, G3; IgG3, 2b; IgG2b, 2a; IgG2a, G1; IgG1, G4; IgG4, G2; IgG2, A; IgA, M; IgM, λ; lambda light chain, κ; kappa light chain. (B) Epitope mapping of ApoB100 affinity-purified mouse and human antibodies as well as of mouse monoclonal antibody 22B4 raised against pB1. The mouse sera used for affinity purification were harvested and pooled at the end of the experiments (33 weeks of age) from Chow-fed or HFD-fed mice. The human sera were pooled after the first ELISA screening experiments and also affinity-purified before the dot blot analysis. 0.2 µg of the given antibody preparation were added per well. a, b and c indicate the amounts of the peptides or proteins spotted per well; a, b and c = 5, 2.5 and 1 µg for p45, p210, p240, pB1 and BSA (bovine serum albumin; negative control); a, b and c = 5, 2.5 and 1 ng for immunoglobulins (M/H IgG = mouse or human IgG; positive control), respectively.