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 isolated antibody having an affinity to the N-terminal portion of PTH (1-84), the method comprising the steps:
a) administering rat (1-34) PTH peptide to a host animal to induce antibody production against said rat (1-34) PTH peptide in said host animal;
b) monitoring antibody titer produced by said administration of said rat (1-34) PTH peptide to said host animal;
c) extracting antisera produced in said host animal by said administration of said rat (1-34) PTH peptide; and
d) isolating and selecting said antibody from said antisera extracted in step c) by affinity chromatography utilizing a peptide antigen selected from the group consisting of SEQ ID NO. 3 and SEQ ID NO. 4.
2. The method of claim 1 further including the following step:
e) attaching a detectable moiety to the antibody isolated and selected in step d).
3. The method of claim 2 wherein said detectable moiety is selected from the group consisting of a radioactive tracer, a fluorescent tracer, an enzymatic tracer and a dye tracer.
4. The method of claim 1 wherein the host animal is a goat.
5. A method for producing an isolated antibody having an affinity to the N-terminal portion of PTH (1-84), the method comprising the steps:
a) administering human (1-34) PTH peptide to a host animal to induce antibody production against said human (1-34) PTH peptide in said host animal;
b) monitoring antibody titer produced by said administration of said human (1-34) PTH peptide to said host animal;
c) extracting antisera produced in said host animal by said administration of said human (1-34) PTH peptide; and
d) isolating and selecting said antibody from said antisera extracted in step c) by affinity chromatography utilizing a peptide antigen selected from the group consisting of SEQ ID NO. 5 and SEQ ID NO. 6.
6. The method of claim 5 further including the following step:
e) attaching a detectable moiety to the antibody isolated and selected in step d).
7. The method of claim 6 wherein said detectable moiety is selected from the group consisting of a radioactive tracer, a fluorescent tracer, an enzymatic tracer and a dye tracer.
8. The method of claim 5 wherein the host animal is a goat.