Quantitation of a pregnancy associated globulin in the rat by two-site binding enzyme immunoassay.

An indirect two-site binding enzyme immunoassay has been developed to quantify the pregnancy associated globulin (PAG) in rats. Anti-PAG from guinea pig is adsorbed to solid phase and binds serum-PAG, which then reacts with its remaining free determinants with anti-PAG from rabbit. Sheep anti-rabbit IgG, labeled with horse radish peroxidase (HRP), binds to the rabbit anti-PAG antibody and the enzymic activity of the solid phase is measured by incubation with the appropriate chromogenic substrate. The optical density is directly related to the quantity of PAG to be measured. Reproducible results are obtained in the range from 5 to 200 micrograms/l. The test detects 12 fmoles of PAG. Female rats show a 1000-fold higher serum level of PAG than male rats. Strain differences of the distribution of the PAG serum concentration were only found between male rats.