Enzyme immunoassay of testosterone in plasma, with use of polyethylene glycol to separate antibody-bound and free hormone.

We describe an enzyme immunoassay for testosterone in which we use a testosterone-3-(carboxymethyl)oxime horseradish peroxidase conjugate as the label and an antiserum, raised in rabbits, to testosterone-3-(carboxymethyl)oxime-bovine serum albumin conjugate. Polyethylene glycol (Carbowax 6000) is used to separate antibody-bound and free steroid. The assay has a sensitivity of 12 pg/assay tube and satisfies the usual criteria of specificity, precision and accuracy. The results agree well with those obtained with a comparable radioimmunoassay.