Sulfonated Sclerostin, antibodies, epitopes and methods for identification and use therefor

A method for production of an antibody specific for a sulfonation site in Sclerostin, comprising the steps of:

• (a) immunizing an animal with sulfonated Sclerostin or sulfonated fragments thereof;
• (b) isolating antibody producing cells from said immunized animal;
• (c) carrying out a screening process with said antibody producing cells to identify cells which produce antibodies specific for Sclerostin or fragments thereof;
• and
• (d) carrying out an additional screening on cells identified in step (c) to identify cells which produce antibodies that discriminate between sulfonated and unsulfonated forms of Sclerostin.

A method for production of an antibody specific for a sulfonation site in Sclerostin, comprising the steps of:

• (a) immunizing an animal with a peptide derived from the Sclerostin amino acid sequence, wherein said peptide comprises a sulfonation site in said Sclerostin;
• (b) isolating antibody producing cells from said immunized animal;
• (c) carrying out a screening process with said antibody producing cells to identify cells which produce antibodies specific for Sclerostin or fragments thereof;
• and
• (d) carrying out an additional screening on cells identified in step (c) to identify cells which produce antibodies that discriminate between sulfonated and unsulfonated forms of Sclerostin.

A method for identification of an antibody specific for a sulfonation site in Sclerostin, comprising the steps of:

• (a) providing a library of antibody producing cells;
• (b) carrying out a screening process with said antibody producing cells to identify cells which produce antibodies specific for Sclerostin or fragments thereof;
• and
• (c) carrying out an additional screening on cells identified in step (b) to identify cells which produce antibodies that discriminate between sulfonated and unsulfonated forms of Sclerostin.

A method for production of an antibody specific for a dimer of Sclerostin, comprising the steps of:

• (a) immunizing an animal with dimerized Sclerostin or dimerized Sclerostin fragments thereof;
• (b) isolating antibody producing cells from said immunized animal;
• and
• (c) carrying out a screening process with said antibody producing cells to identify cells which produce antibodies that discriminate between dimerized and undimerized forms of Sclerostin.

A method of identifying compounds which disrupt or inhibit sulfonation of Sclerostin, comprising the steps of:

• (a) providing:
• (i) a library of compounds to be tested;
• (ii) Sclerostin wherein sulfonation modification sites in said Sclerostin remain unsulfonated;
• (iii) TPST;
• and
• (iv) PAPS;
• (b) forming a mixture comprising said Sclerostin (ii), TPST (iii), PAPS (iv) and a compound from said library (i);
• and
• (c) measuring the amount of conversion of unsulfonated Sclerostin into sulfonated Sclerostin, thereby identifying whether said compound disrupts or inhibits said sulfonation of Sclerostin.

A method of identifying compounds which disrupt or inhibit sulfonation of Sclerostin, comprising the steps of:

• (a) providing:
• (i) a virtual model of the structure of Sclerostin;
• (ii) a virtual library of compounds to be tested;
• (iii) Sclerostin wherein sulfonation modification sites in said Sclerostin remain unsulfonated;
• (iv) TPST;
• and
• (v) PAPS;
• (b) carrying out virtual screening processes to identify compounds from said library (ii) that would bind to a sulfonation modification site in Sclerostin;
• (c) forming a mixture comprising said Sclerostin (iii), said TPST (iv), said PAPS (v) and said compound identified by step (b);
• and
• (d) measuring the amount of conversion of unsulfonated Sclerostin into sulfonated Sclerostin, thereby identifying whether said compound disrupts or inhibits said sulfonation of Sclerostin.

A method for identification of a compound which disrupts or inhibits the interaction between Sclerostin and a binding partner, comprising the steps of:

• (a) providing;
• (i) a library of compounds to be tested;
• (ii) a composition comprising a peptide which comprises a sulfonated epitope of Sclerostin;
• and
• (iii) said binding partner for Sclerostin;
• (b) forming a mixture comprising said composition (ii), said binding partner (iii) and a compound from said library (i);
• and
• (c) measuring the effect of said compound from the library (i) on the ability to disrupt or inhibit the binding between said composition (ii) and said binding partner (iii), thereby identifying whether said compound from the library (i) disrupts or inhibits the interaction between Sclerostin and its binding partner.

A method for identification of a compound which disrupts or inhibits the interaction between Sclerostin and a binding partner, comprising the steps of:

• (a) providing;
• (i) a library of compounds to be tested;
• (ii) a composition comprising a peptide which comprises an unsulfonated epitope of the sulfonation modification site of Sclerostin and
• (iii) said binding partner for Sclerostin;
• (b) forming a mixture comprising said composition (ii), said binding partner (iii) and a compound from said library (i);
• (c) measuring the effect of said compound from said library (i) on the ability to disrupt or inhibit the binding between said composition (ii) and said binding partner (iii), thereby to identify whether said compound from said library (i) disrupts or inhibits the interaction between Sclerostin and its binding partner.

A method of identifying compounds which disrupt or inhibit interaction between Sclerostin and a binding partner, comprising the steps of:

• (a) providing:
• (i) a virtual model of the structure of Sclerostin that includes a sulfonation modification of one or more amino acids;
• (ii) a virtual library of compounds to be tested;
• (iii) sulfonated Sclerostin;
• (iv) a binding partner for said sulfonated Sclerostin (iii);
• and
• (v) reagents for a biological assay that is dependent upon an interaction between Sclerostin and a binding partner;
• (b) carrying out virtual screening processes to identify compounds from said library (ii) that would bind to a sulfonation modification site in Sclerostin;
• (c) obtaining the compound or compounds identified by carrying out the virtual screening processes in step (b);
• (c) forming a mixture comprising said sulfonated Sclerostin (iii), said binding partner (iv), said reagents for a biological assay (v) and said compounds or compounds obtained in step (c);
• and
• (d) carrying out said biological assay to identify whether said compound or compounds obtained in step (c) and from said library (i) disrupt or inhibit the interaction of said sulfonated Sclerostin and its binding partner.