Single Step Purification of Ovine Luteinizing Hormone by Affinity Chromatography

• Published in: The Journal of biological chemistry Vol. 247; no. 20; pp. 6491 - 6498
• Main Author: Gospodarowicz, Denis
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 25.10.1972; American Society for Biochemistry and Molecular Biology
• Subjects: Amino Acids - analysis; Animals; Cattle; Chemical Phenomena; Chemistry; Chromatography, Affinity; Cyanogen Bromide; Electrophoresis, Disc; Fetus; Guanidines; Hexosamines - analysis; Horses; Immunodiffusion; Immunoelectrophoresis; Immunoglobulin G; Iodine Isotopes; Luteinizing Hormone - analysis; Luteinizing Hormone - blood; Luteinizing Hormone - isolation & purification; Pituitary Gland; Polysaccharides; Precipitin Tests; Rabbits - immunology; Radioimmunoassay; Sheep
• ISSN: 0021-9258; 1083-351X
• DOI: 10.1016/S0021-9258(19)44719-4

Affinity chromatography has been applied to the problem of specifically removing luteinizing hormone from sera used to supplement tissue culture media. Horse or fetal calf sera, passed through a column of an anti-luteinizing hormone immunoglobulin fraction linked covalently to agarose, were specifically depleted in their content of luteinizing hormone as seen by radioimmunoassay and by their effect on cultures of ovarian cells dependant on luteinizing hormone for their growth. Affinity chromatography has also been applied to the one step isolation of luteinizing hormone from a partially purified or crude extract of ovine pituitary glands. Partially purified extracts or crude extracts of pituitary glands, passed through a column of an anti-luteinizing hormone immunoglobulin fraction linked covalently to agarose, were depleted in their luteinizing hormone content. Elution of the column by 6 m guanidine HCl pH 1.5 resulted in the recovery of the hormone which was further characterized by chemical and biological analysis as well as its electrophoretic pattern. It was demonstrated that affinity chromatography can be used to isolate preparations of highly homogeneous ovine luteinizing hormone in one step.