Human Anti-TSHR Recombinant Antibody (clone K1-70) (CAT#: PABL-737)

Figure 1 Crystal structure of TSHR260 in complex with thyroid stimulating monoclonal autoantibody (M22) Fab (TSHR-M22) and with the TSHR blocking monoclonal autoantibody (K1-70) Fab (TSHR-K1-70).

Cartoon diagrams of the complex structures are shown. TSHR is in cyan, light chains are in green and the heavy chains in blue. Disulphide-bonded cysteines are shown as balls and sticks, disulphide bonds are in yellow and cysteine residues are labelled in orange. The N- and C-termini are marked. M22 interactions with TSHR amino acid R255 are marked and shown as balls and sticks. In contrast there are no interactions between K1-70 and amino acid R255 or the C-terminus of the TSHR. Reproduced from Figure 5 of Furmaniak et al. [3] with kind permission from Springer Science and Business Media.

Furmaniak, J., Sanders, J., Miguel, R. N., & Smith, B. R. (2015). Mechanisms of action of TSHR autoantibodies. Hormone and Metabolic Research, 47(10), 735-752.

Figure 2 Bridge type TRAb ELISA using TSHR 260 alkaline phosphatase.

A. Binding of stimulating (K1-18 and M22) and blocking (K1-70) human monoclonal autoantibodies and B binding of K1 donor serum IgG (lower x-axis); NIBSC 90/672 (upper x-axis). K1-18 IgG (♦); K1-70 IgG (▪); M22 IgG (▴); control MAb (5B3 IgG) (×); K1 donor serum (□); NIBSC 90/672 (○).

Smith, B. R., Sanders, J., Evans, M., Tagami, T., & Furmaniak, J. (2009). TSH receptor–autoantibody interactions. Hormone and Metabolic Research, 41(06), 448-455.

Figure 3 Inhibition of porcine TSH (3 ng/mL)-mediated stimulation of cyclic AMP production by TSHR monoclonal antibodies with antagonist activity.

MAb-B2 IgG (m); K1–70 IgG (d); 5C9 IgG (■).

Sanders, J., Miguel, R. N., Furmaniak, J., & Smith, B. R. (2010). TSH receptor monoclonal antibodies with agonist, antagonist, and inverse agonist activities. In Methods in enzymology (Vol. 485, pp. 393-420). Academic Press.

Figure 3 Inhibition of porcine TSH (4 ng/mL)-mediated stimulation of cyclic AMP production by TSHR monoclonal antibodies with antagonist activity.

(A)Human MAb 5C9 (antagonist and inverse agonist; derived from the comparative model of 5C9 structure), (B) mouse MAb-B2 (antagonist; derived from the crystal structure solved at 3.3 A˚ resolution), (C) human MAb K1–70 (antagonist; derived from the crystal structure solved at 2.2 A˚ resolution), (D) human MAb M22 (agonist; derived from the crystal structure solved at 1.65 A˚ resolution). Position of the charged residues is indicated in the figures on the left-hand side of the panel and the position of aromatic
residues is indicated in the figures on the right-hand side of the panel. Acidic patches are shown in red and basic patches in blue (See Color Insert).

Sanders, J., Miguel, R. N., Furmaniak, J., & Smith, B. R. (2010). TSH receptor monoclonal antibodies with agonist, antagonist, and inverse agonist activities. In Methods in enzymology (Vol. 485, pp. 393-420). Academic Press.

Figure 1 Human Anti-TSHR Recombinant Antibody (PABL-737) in SDS-PAGE

SDS-PAGE analysis of PABL-737: Non-Reducing (Lane 1, 1.0 µg) and Reducing (Lane 2, 2.0 µg)