OB polypeptide antibodies and method of making

An antibody to an OB polypeptide, the polypeptide comprising:

• a) amino acid sequence set out in SEQ ID NO:
• 2;
• b) the amino acid sequence set out in amino acids 22-167 of SEQ ID NO:
• 2;
• c) the amino acid sequence set out in SEQ ID NO:
• 4;
• or
• d) the amino acid sequence set out in amino acids 22-167 of SEQ ID NO:
• 4.

An antibody to an OB polypeptide, the polypeptide comprising:

• a) the amino acid sequence set out in amino acids 22-167 of SEQ ID NO:
• 2 having an N-terminal methionine or an N-terminal polyhistidine;
• or
• b) the amino acid sequence set out in amino acids 22-167 of SEQ ID NO:
• 4 having an N-terminal methionine or an N-terminal polyhistidine.

An antibody to an OB polypeptide, the polypeptide comprising:

• a) the amino acid sequence set out in SEQ ID NO:
• 5;
• b) the amino acid sequence set out in amino acids 22-166 of SEQ ID NO:
• 5;
• c) the amino acid sequence set out in amino acids 22-166 of SEQ ID NO:
• 5, having an N-terminal methionine or an N-terminal polyhistidine;
• d) the amino acid sequence set out in SEQ ID NO:
• 6;
• e) the amino acid sequence set out in amino acids 22-166 of SEQ ID NO:
• 6;
• or
• f) the amino acid sequence set out in amino acids 22-166 of SEQ ID NO:
• 6 having an N-terminal methionine or an N-terminal polyhistidine.

An antibody to an immunogenic fragment of an OB polypeptide selected from the group consisting of the polypeptide of SEQ ID NO:

• 18, the polypeptide of SEQ ID NO:
• 19, the polypeptide of SEQ ID NO:
• 20 and the polypeptide of SEQ ID NO:
• 21.

A method for preparing an antibody specific to an OB polypeptide, comprising:

• (a) conjugating to a carrier protein a mammalian OB polypeptide, said polypeptide having the sequence of a naturally occurring mammalian OB polypeptide, having as a mature protein about 145 amino acids, and capable of modulating body weight;
• (b) immunizing a host animal with the OB polypeptide fragment-carrier protein conjugate of step (a) admixed with an adjuvant;
• and
• (c) obtaining antibody from the immunized host animal.

A method for preparing an antibody specific to an OB polypeptide, comprising:

• (a) conjugating to a carrier protein an OB polypeptide comprising:
• i) the amino acid sequence set out in SEQ ID NO:
• 2;
• ii) the amino acid sequence set out in amino acids 22-167 of SEQ ID NO:
• 2;
• iii) the amino acid sequence set out in SEQ ID NO:
• 4;
• or
• iv) the amino acid sequence set out in amino acids 22-167 of SEQ ID NO:
• 4;
• (b) immunizing a host animal with the OB polypeptide fragment-carrier protein conjugate of step (a) admixed with an adjuvant;
• and
• (c) obtaining antibody from the immunized host animal.

A method for preparing an antibody specific to an OB polypeptide, comprising:

• (a) conjugating to a carrier protein an OB polypeptide comprising:
• i) the amino acid sequence set out in SEQ ID NO:
• 5;
• ii) the amino acid sequence set out in amino acids 22-166 of SEQ ID NO:
• 5;
• iii) the amino acid sequence set out in SEQ ID NO:
• 6;
• or
• iv) the amino acid sequence set out in amino acids 22-166 of SEQ ID NO:
• 6;
• (b) immunizing a host animal with the OB polypeptide fragment-carrier protein conjugate of step (a) admixed with an adjuvant;
• and
• (c) obtaining antibody from the immunized host animal.

A method for preparing an antibody specific to an OB polypeptide, comprising:

• (a) conjugating to a carrier protein an immunogenic fragment of an OB polypeptide;
• (b) immunizing a host animal with the OB polypeptide fragment-carrier protein conjugate of step (a) admixed with an adjuvant;
• and
• (c) obtaining antibody from the immunized host animal.

The method of according to claim wherein the immunogenic fragment is selected from the group of the polypeptide of SEQ ID NO:

• 18, the polypeptide of SEQ ID NO:
• 19, the polypeptide of SEQ ID NO:
• 20, and the polypeptide of SEQ ID NO:
• 21.

A method for preparing an antibody specific to an OB polypeptide, comprising:

• (a) conjugating to a carrier protein an OB polypeptide comprising amino acids 22-167 of SEQ ID NO:
• 4 wherein one or more amino acids selected from the group consisting of amino acids 53, 56, 71, 85, 89, 92, 95, 98, 110, 118, 121, 122, 126, 127, 128, 129, 132, 139, 157, 159, 163 and 166 is substituted with a conserved amino acid;
• (b) immunizing a host animal with the OB polypeptide fragment-carrier protein conjugate of step (a) admixed with an adjuvant;
• and
• (c) obtaining antibody from the immunized host animal.

A method for preparing an antibody specific to an OB polypeptide, comprising:

• (a) conjugating to a carrier protein an OB polypeptide comprising amino acids 22-167 of SEQ ID NO:
• 4 wherein one or more amino acids selected from the group consisting of amino acids 53, 56, 71, 85, 89, 92, 95, 98, 110, 118, 121, 122, 126, 127, 128, 129, 132, 139, 157, 159, 163 and 166 is substituted with the particular amino acid present at the corresponding position in SEQ ID NO:
• 2;
• (b) immunizing a host animal with the OB polypeptide fragment-carrier protein conjugate of step (a) admixed with an adjuvant;
• and
• (c) obtaining antibody from the immunized host animal.

A method for preparing an antibody specific to an OB polypeptide, comprising:

• (a) conjugating to a carrier protein an OB polypeptide having 83 percent or greater amino acid sequence identity to the OB polypeptide amino acid sequence set out in SEQ ID NOS:
• 2, 4, 5 or 6;
• (b) immunizing a host animal with the OB polypeptide fragment-carrier protein conjugate of step (a) admixed with an adjuvant;
• and
• (c) obtaining antibody from the immunized host animal.

A test kit for determining the presence or amount of OB polypeptide comprising:

• (a) a predetermined amount of at least one labeled antibody to an OB polypeptide obtained by the direct or indirect attachment of an antibody to an OB polypeptide to a detectable label;
• (b) other reagents;
• and
• (c) directions for use of said kit.

A test kit for determining the presence or amount of OB polypeptide comprising:

• (a) a labeled component which has been obtained by coupling an OB polypeptide to a detectable label;
• (b) one or more additional immunochemically reactive components of which at least one component is an antibody to an OB polypeptide;
• and
• (c) directions for the performance of a protocol for the detection and/or determination of one or more components of an immunochemical reaction between OB polypeptide and an antibody to an OB polypeptide.