The glycoprotein hormones are heterodimeric proteins that share a common alpha subunit and have unique beta subunits that confer receptor selectivity. One member of this family, follicle-stimulating hormone (FSH), is secreted by the pituitary and is involved in the control of male and female reproduction. Herein, we describe the construction of baculoviruses for glutathione-S-transferase (GST) fusions of the human FSH (hFSH) subunits and their expression in insect cells, either alone or with the complementary non-fused FSH subunits (FSHalpha or FSHbeta). Only the GST-BV-hFSHalpha monomer and the GST-BV-hFSHalpha/BV-hFSHbeta (GST-BV-hFSH) heterodimer were efficiently secreted into the culture supernatant. The hybrid molecule, GST-BV-hFSH, was affinity purified in one step, and demonstrated activity in receptor-radioligand binding assays and in a cAMP accumulation assay. The use of GST-BV-hFSHalpha provides a novel and efficient method for purifying and studying members of the glycoprotein hormone family derived from the culture supernatant or subcellular fractions of the cell. (C) 2004 Elsevier Inc. All rights reserved.