Influence of bovine LH tracer quality on levels of LH in GnRH-treated cows.

Chromatography of 125I-bovine LH (LER-1716-2 and USDA-I-1) by means of anion exchange high performance liquid chromatography (HPLC) revealed two main peaks of radioactivity regardless as to whether or not the tracer was initially purified on cellulose CF11. The content of radioactivity in the first peak tended to increase as the storage time of the bLH preparation, either before or after iodination, increased. The first peak of radioactivity after HPLC fractionation either with or without cellulose adsorption consisted of material with low binding ability to bLH antiserum (6.9% +/- 0.5 and 13.0% +/- 1.0, respectively) and high binding ability to ovine LH alpha antiserum (51.0% +/- 2.7 and 35.2% +/- 3.6, respectively). The average ratio of alpha-subunit immuno-reactivity to 125I-bLH immunoreactivity in this material was 7.4 +/- 0.1 and 2.7 +/- 0.2, respectively (P less than 0.001). Peaks in 125I-bLH radioactivity and 125I-bLH immunoreactivity had different elution times. Radioimmunoassays with tracers obtained from fractions derived from the first radioactive peak after HPLC chromatography (i.e. 125I-bLH-LER-1716-2) both with and without cellulose adsorption, yielded significantly lower mean plasma LH levels in GnRH-treated cows compared with the control tracer routinely purified only on cellulose CF11 (e.g. 5.7 vs. 8.2 micrograms/; 4.6 vs. 8.2 micrograms/l). Plasma LH levels in GnRH-treated cows were significantly (P less than 0.001) lower as measured by radioimmunoassay utilizing 125I-USDA-blH-I-1 tracers than by radioimmunoassays utilizing 125I-blH-LER-1716-2 tracers (i.e. either Y = 0.17 + 0.75X or Y = 1.18 + 0.60X).