Antibodies and peptide antigens for producing antibodies having a selective binding specificity to bioactive intact parathyroid hormone (PTH) 1-84
View Patent ↗Peptide antigens corresponding to amino acid residues 2-12, 1-12, 2-15 and 1-15 of parathyroid hormone (PTH), antibodies having an affinity to such peptide antigens and methods of producing the same. Such antigens, antibodies and methods producing the same according to the present invention are useful in determining bioactive intact PTH levels in serum, plasma, and/or cell culture media. Such antibodies further possess a high degree of species cross-reactivity, but substantially mitigated cross-reactivity to non-whole PTH peptide fragments and little to no recognition of the first amino acid residue of PTH.
1. A method for producing an antibody to the N-terminal portion of (1-84) PTH useful in the determination of intact PTH 1-84 levels in a biological sample, the method comprising the steps:
a) administering a first peptide antigen to a host animal to induce antibody production against said first peptide antigen in said host animal, said first peptide antigen being selected from the group consisting of SEQ ID NO. 3, SEQ ID NO. 4, SEQ ID NO. 5, SEQ ID NO. 6, (1-34) PTH and (1-84) PTH;
b) monitoring antibody titer produced by said administration of said first antigen to said host animal;
c) extracting antisera produced in said host animal by said administration of said first peptide antigen;
d) isolating and selecting at least one antibody from said antisera extracted in step c) by affinity chromatography utilizing a second peptide antigen selected from the group consisting of SEQ ID NO. 3, SEQ ID NO. 4, SEQ ID NO. 5, and SEQ ID NO.6;
e) removing said at least one antibody isolated in step d) having specificity for a peptide antigen selected from the group consisting of (4-34) PTH, (5-34) PTH, (4-84) PTH and (5-84) PTH; and
f) isolating said antibody in steps d) and e) having a binding affinity for at most the first three or four amino acid residues of the N-terminal portion of PTH and wherein the isolated antibody has no binding affinity for any amino acid residues at or beyond the fifth amino acid residues of the N-terminal of PTH.
2. The method of claim 1 wherein in step a), said host animal is selected from the group consisting of a mouse and a rabbit.
3. The method of claim 1 wherein in step a), said host animal comprises at least one goat.
4. The method of claim 1 wherein in step a), said (1-34) PTH is from a species selected from the group consisting of a human, a rat, a mouse, a bovine, a dog, a pig, a cat, and a monkey.
5. The method of claim 1 wherein in step a), said (1-84) PTH is from a species selected from the group consisting of a human, a rat, a mouse, a bovine, a dog, a pig, a cat, and a monkey.