A simple method for the isolation of pregnant mare serum gonadotropin.

This research devised a straightforward method for purifying pregnant mare serum gonadotropin (PMSG), a hormone produced by pregnant mares, attaining an average specific activity of 15,800 IU/mg and yield rates of 50-80%. The purified PMSG was then scrutinized for biological activity, column electrophoresis, and ultracentrifugation.

Purification of Pregnant Mare Serum Gonadotropin

• In this research, the aim was to develop a simple process for the purification of pregnant mare serum gonadotropin (PMSG). PMSG is a hormone produced by the endometrium of pregnant mares and has both FSH-like (Follicle-stimulating hormone) and LH-like (Luteinizing hormone) activities, making it crucial in equine reproduction.
• The purification was achieved using pH fractionation, which involves separating the various components based on their pH levels. A substance known as metaphosphoric acid was used for this purpose. This was followed by alcohol precipitation, a common lab method used to concentrate or isolate proteins.

Gel-Filtration and Chromatography

• After isolation, the study further purified PMSG using gel-filtration on Sephadex G-100, a type of size exclusion chromatography. This type of chromatography separates molecules based on their size, allowing larger molecules to pass through the column faster than smaller ones.
• The study also used chromatography on sulfoethyl-Sephadex C-50, another filtration method. Here, the components are separated based on their different affinities for the chromatography resin used (sulfoethyl-Sephadex in this case).

Purity and Yield

• The final PMSG product obtained had an average specific activity of 15,800 IU/mg. This is a measurement that shows the potency or activity level of an enzyme or hormone per milligram of total protein.
• The technique demonstrated effective purification of PMSG, with an efficiency rate of 50-80%. This means that 50-80% of the PMSG that was initially present in the unfractionated serum was recovered in the purified product.

Characterization of the Purified Product

• Upon successful purification of PMSG, the study characterized the product with respect to its biological activity, which means the effect it had on a living organism or biological process.
• The research further analyzed the product using column electrophoresis and ultracentrifugation.
• Electrophoresis on columns of polyacrylamide allows for the separation of macromolecules like proteins based on their electrical charge and size.
• Ultracentrifugation is a process where samples are spun at very high speeds to separate particles based on their density and size.