Epitope regions of a thyrotrophin (TSH) receptor, uses thereof and antibodies thereto

A method of screening a sample of body fluid for TSH receptor autoantibodies wherein said sample is from a subject suspected of suffering from, susceptible to, having or recovering from autoimmune disease associated with an immune reaction to a TSH receptor, said method comprising:

• (a) providing said sample of body fluid from said subject;
• (b) contacting said sample with
• (i) a full length TSH receptor comprising amino acid residues 246-260, wherein the amino acid sequence of residues 246-260 comprises amino acid residues 47 to 61 of SEQ ID NO:17 or 22, and
• (ii) one or more murine monoclonal antibodies capable of competing with autoantibodies to a TSH receptor in the binding of said full length TSH receptor, wherein said one or more competitor monoclonal antibodies is positive for thyroid stimulating activity at a concentration of 20 μg/ml in a cyclic AMP thyroid cell assay and inhibits binding of TSH to TSH receptor, said one or more monoclonal antibodies being classified as positive if the thyroid stimulating activity of said one or more monoclonal antibodies is greater than 180 percent in the cyclic AMP thyroid cell assay, said percent thyroid stimulating activity comprising 100× ratio of cyclic AMP produced in the presence of said one or more monoclonal competitor antibodies to cyclic AMP produced in the presence of sera pooled from healthy blood donors,
• so as to permit said full length TSH receptor with either autoantibodies to TSH receptor present in said sample, or said one or more competitor monoclonal antibodies;
• and
• (c) detecting the binding of said full length TSH receptor with said autoantibodies present in said sample, thereby providing an indication of the presence of said autoantibodies to a TSH receptor in said sample.

A method of screening a sample of body fluid for autoantibodies to a TSH receptor wherein said sample is from a subject suspected of suffering from, susceptible to, having or recovering from autoimmune disease associated with an immune reaction to a TSH receptor, said method comprising:

• (a) contacting said sample with
• (i) a full length TSH receptor, one or more epitopes thereof or a and
• (ii) one or more murine monoclonal antibodies for the TSH receptor, wherein said one or more monoclonal antibodies:
• does not comprise naturally produced autoantibodies to the TSH receptor,
• inhibits binding of TSH to TSH receptor, and
• is positive for thyroid stimulating activity at a concentration of 20 μg/ml in a cyclic AMP thyroid cell assay, said one or more monoclonal antibodies being classified as positive if the thyroid stimulating activity of said one or more monoclonal antibodies is greater than 180 percent in the cyclic AMP thyroid cell assay, wherein said percent thyroid stimulating activity comprises 100× ratio of cyclic AMP produced in the presence of said one or more monoclonal antibodies to cyclic AMP produced in the presence of sera pooled from healthy blood donors;
• so as to permit said TSH receptor to bind with either autoantibodies to TSH receptor present in said sample, or said one or more monoclonal antibodies for the TSH receptor;
• and
• (b) detecting binding of said TSH receptor, with said autoantibodies present in said sample, thereby providing an indication of the presence of said autoantibodies to TSH receptor in said sample.

A method of screening a sample of body fluid for TSH receptor autoantibodies wherein said sample is from a subject suspected of suffering from, susceptible to, having or recovering from autoimmune disease associated with an immune reaction to a TSH receptor, said method comprising:

• (a) contacting said sample with
• (i) a full length TSH receptor, and
• (ii) one or more murine monoclonal antibodies for said TSH receptor, wherein said one or more monoclonal antibodies is positive for thyroid stimulating activity at a concentration of 20 μg/ml in a cyclic AMP thyroid cell assay and inhibits binding of TSH to TSH receptor, said one or more monoclonal antibodies being classified as positive if the thyroid stimulating activity of said one or more monoclonal antibodies is greater than 180 percent in the cyclic AMP thyroid cell assay, said percent thyroid stimulating activity comprising 100× ratio of cyclic AMP produced in the presence of said one or more monoclonal antibodies to cyclic AMP produced in the presence of sera pooled from healthy blood donors;
• so as to permit said TSH receptor to bind with either autoantibodies to said TSH receptor present in said sample, or said one or more monoclonal antibodies;
• and
• (b) detecting binding of said TSH receptor with said autoantibodies present in said sample, thereby providing an indication of the presence of said autoantibodies to TSH receptor in said sample;
• wherein said one or more monoclonal antibodies for said TSH receptor are directly or indirectly immobilised to a surface either prior to, or after step (a).