Human Anti-TGM2 Recombinant Antibody (clone 679-14-E06) (CAT#: PABL-735)

Figure 1 Binding of a panel of monoclonal antibodies to TG2 mutants as assessed by ELISA.

Binding of a panel of antibodies reactive with epitope 1, 2, 3, or 4 was tested against the TG2 mutants R19S, K30A, R116A, and H134A. Shown are signals normalized against those obtained with WT TG2. The open circles represent an anti-TG2 antibody that was not assigned to epitope 1– 4, and diamonds represent antibodies analyzed in the SPR study. The symbols for the two epitope 1 antibodies 679-14-E06 and 693-10-B06 are shown in red and blue, respectively. Symbols representing antibodies 693-1-A03 and 763-
4-B06 recognizing epitope 2 are colored yellow and green. Symbols for the epitope 3 antibodies 763-4-A06 and 763-4-E05 are shown in orange and gray, and symbols for 693-1-D03, which recognizes epitope 4, are shown in violet. Horizontal lines indicate medians. Results from one of three independent experiments are shown.

Chen, X., Hnida, K., Graewert, M. A., Andersen, J. T., Iversen, R., Tuukkanen, A., ... & Sollid, L. M. (2015). Structural basis for antigen recognition by transglutaminase 2-specific autoantibodies in celiac disease. Journal of Biological Chemistry, jbc-M115.

Figure 2 Both heavy and light chain of 679-14-E06 are involved in binding of epitope 1.

Heavy and light chains were swapped between antibodies reactive (679-14-E06) and not reactive (679-14-A04) to TG2. A, Coomassie-stained SDS-PAGE gels showing 679-14-E06H/679-14-A04L and 679-14-A04H/679-14-E06L antibodies under reducing and non-reducing conditions (white lines show the borders of different lanes). B, reactivity of the antibodies 679-14-E06, 679-14-A04, 679-14-E06H/679-14-A04L, and 679-14-A04H/679-14-E06L to TG2 as monitored by ELISA. Shown are the results from one of two independent
experiments.

Chen, X., Hnida, K., Graewert, M. A., Andersen, J. T., Iversen, R., Tuukkanen, A., ... & Sollid, L. M. (2015). Structural basis for antigen recognition by transglutaminase 2-specific autoantibodies in celiac disease. Journal of Biological Chemistry, jbc-M115.

Figure 3 Involvement of the TG2 residues Asp-191 and Lys-265 in the epitope of 679-14-E06 and other epitope 1 antibodies.

A and B, binding of antibody 679-14-E06 to the TG2 mutants D191A and K265A as assessed by ELISA and SPR. The ELISA and SPR experiments were done at least twice. C, testing of the binding to the TG2 mutant D191A by a panel of anti-TG2 antibodies reactive with epitope 1, 2, 3, or 4. Shown are signals normalized against those obtained with WT TG2. The gray point represents the antibody 679-14-E06, and the open circles represent anti-TG2 antibodies that were not assigned to epitope 1-4. Horizontal lines indicate medians.

Chen, X., Hnida, K., Graewert, M. A., Andersen, J. T., Iversen, R., Tuukkanen, A., ... & Sollid, L. M. (2015). Structural basis for antigen recognition by transglutaminase 2-specific autoantibodies in celiac disease. Journal of Biological Chemistry, jbc-M115.

Figure 4 Characterization of TG2 ECM interaction.

(A) Colocalization analysis of TG2 and ECM proteins as shown in Fig. 4. The graph shows the Pearson's correlation coefficient calculated as described in the Materials and Methods section. (100% colocalization = 1, no colocalization = 0). The graph show values for all regions analyzed, including mean and SD as error bars. (B) Treatment of tissue sections with the collagenase blend H abrogates binding of full-length TG2 to the tissue ECM as observed with epitope 1 mAb (679-14-E06), whereas collagen VI staining is not influenced.

Chen, X., Hnida, K., Graewert, M. A., Andersen, J. T., Iversen, R., Tuukkanen, A., ... & Sollid, L. M. (2015). Structural basis for antigen recognition by transglutaminase 2-specific autoantibodies in celiac disease. Journal of Biological Chemistry, jbc-M115.