Purification of glycosaminoglycans from bovine follicular fluid

The follicular glycosaminoglycans, dermatan sulfate and heparan sulfate, are not available in sufficient quantities for detailed analytical studies or to use as reagents in cell cultures. The purpose of this study was to purify follicular glycosaminoglycans simultaneously from bovine follicular flui...

Full description

Saved in:
Bibliographic Details
Published in:Journal of dairy science 1987-09, Vol.70 (9), p.1913-1919
Main Authors: Bellin, M.E, Ax, R.L
Format: Article
Language:English
Subjects:
Animals
BODY FLUIDS
Body Fluids - analysis
BOVIN
CATTLE
Cattle - physiology
Chondroitin - analogs & derivatives
CHROMATOGRAPHIE
CHROMATOGRAPHY
CROMATOGRAFIA
dermatan sulfate
Dermatan Sulfate - isolation & purification
EPURATION
Female
GANADO BOVINO
Glycosaminoglycans - isolation & purification
heparan sulfate
Heparitin Sulfate - isolation & purification
LIQUIDE BIOLOGIQUE
LIQUIDOS CORPORALES
Ovarian Follicle - analysis
POLISACARIDOS
POLYHOLOSIDE
POLYSACCHARIDES
PURIFICACION
PURIFICATION
SULFATE
SULFATOS
SULPHATES
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:The follicular glycosaminoglycans, dermatan sulfate and heparan sulfate, are not available in sufficient quantities for detailed analytical studies or to use as reagents in cell cultures. The purpose of this study was to purify follicular glycosaminoglycans simultaneously from bovine follicular fluid. The use of a chloroform:methanol extraction of 10 ml of fluid resulted in recoveries of at least 90% of the glycosaminoglycans, otherwise an insoluble product resulted. Heparan sulfate and dermatan sulfate eluted at .11 and .46 M NaCl, respectively, when subjected to HPLC with an anion exchange column. Fluid from small follicles allowed to "age" in the ovaries exhibited marked reductions in the concentrations of glycosaminoglycans. Aging of the fluid from large follicles resulted in greater glycosaminoglycan concentrations and appearance of glycosaminoglycans reflecting other charge distributions. Known amounts of purified follicular fluid heparan sulfate and dermatan sulfate were applied to a gel filtration HPLC column, and corresponding integrated peak areas were plotted in relation to the glycosaminoglycan concentrations. Slopes of the lines for commercial chondroitin sulfate and follicular dermatan sulfate were not significantly different, but the slope of the line for follicular heparan sulfate was 13.5-fold greater than the slope for commercial heparin.
ISSN:0022-0302
1525-3198
DOI:10.3168/jds.S0022-0302(87)80231-X