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Synthesis of fluorescein labelled steroid hormone-albumin conjugates for the fluorescent histochemical detection of hormone receptors

1980; Pergamon Press; Volume: 13; Issue: 8 Linguagem: Inglês

10.1016/0022-4731(80)90177-6

1878-2353

Eric Gaetjens, Louis P. Pertschuk,

Monoclonal and Polyclonal Antibodies Research

Estradiol 17β-hemisuccinate, testosterone 17β-hemisuccinate, and progesterone 11α-hemisuccinate were covalently coupled to bovine serum albumin in a stoichiometry of 4–9 mol of hormone per mol of albumin by the isobutylchlorocarbonate mixed anhydride technique. The albumin-hormone conjugates were then labelled with 4–5 mol of fluorescein per mol of albumin by reaction with fluorescein isothiocyanate at pH 9.5 and 0°C for 7 h. Chromatography of the conjugates on Sephadex G-25 under dissociating conditions showed that the preparations contained less than 1% free hormone or fluorescein. These conjugates are currently being utilized for the fluorescent histochemical detection of estrogen, androgen, and progesterone receptors in cancer tissue specimens.