A reference preparation of buffalo pituitary follicle stimulating hormone using lectin affinity chromatography.
An improved and cost effective method to isolate FSH from buffalo pituitary glands is described here. The buFSH activity was monitored throughout by a highly sensitive heterologous radioimmunoassay (sensitivity 0.2 ng oFSH/mL) and the in vivo biological activity of the final preparation was also established. A biologically active buFSH-enriched preparation with a moderate recovery (42%) was obtained. The yield of the final buFSH-enriched preparation was 26.5 mg/kg of buffalo pituitary gland. In SDS-PAGE, the purified buFSH resolved as a heterodimer of 30 kDa molecular size, with a 21 kDa presumptive alpha-subunit. This preparation was also characterized in terms of biological and some of its physicochemical properties. A high-titer antiserum to buFSH was also raised in rabbit using this preparation. The reagents generated, buFSH and buFSH-specific polyclonal antisera, have possible diagnostic and therapeutic usage for improvement of reproductive health of water buffaloes.
Chand
Hitendra S
HS
Hormone Research Laboratory, Department of Zoology, University of Delhi, Delhi, India.
Chaudhary
Rajesh
R
Muralidhar
K
K
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology / Q000302:isolation & purification; D015153:Blotting, Western; D002020:Buffaloes; D002846:Chromatography, Affinity / Q000379:methods; D002850:Chromatography, Gel; D005260:Female; D005640:Follicle Stimulating Hormone / Q000276:immunology / Q000302:isolation & purification* / Q000494:pharmacology; D007106:Immune Sera / Q000276:immunology / Q000302:isolation & purification; D008970:Molecular Weight; D009929:Organ Size / Q000187:drug effects; D010053:Ovary / Q000187:drug effects / Q000254:growth & development; D010902:Pituitary Gland / Q000737:chemistry*; D011817:Rabbits; D011863:Radioimmunoassay; D051381:Rats; D012685:Sepharose / Q000031:analogs & derivatives; D014611:Vaccination
D000911:Antibodies, Monoclonal; D007106:Immune Sera; C028326:concanavalin A-sepharose; D005640:Follicle Stimulating Hormone; D012685:Sepharose