Strategies for purification of four reproductive hormones from the same batch of buffalo (Bubalus bubalis) pituitaries

Abstract

The occurrence of different reproductive hormones like LH, FSH, TSH and prolactin, in different side fractions obtained during the extraction of buffalo pituitary glands either by the procedure of Papkoff et al. [Arch Biochem Biophys, 111 (1965) 431] or by that of Ellis [Endocrinology, 69 (1961) 554], was examined with the aid of antisera to respective heterologous hormones as well as bio-assays. Thus in the procedure of Papkoff et al., the SP-Sephadex fractions could be taken for purification of LH and TSH, while the acid pellet yielded prolactin. Further it was shown that 50% (NH4)2SO4 could be directly size fractionated and following cation exchange chromatography yields LH and TSH. FSH could be purified from 80% ammonium sulphate pellet. In another protocol of Ellis, differential extraction and chromatographic separation yielded all the four reproductive hormones. Some of the physico-chemical and immunobiological characteristics of these hormones are described.