@article {18455,
	title = {Following the enzymatic digestion of chondroitin sulfate by a simple GPC analysis},
	journal = {Analytica Chimica Acta},
	volume = {885},
	year = {2015},
	month = {2015-06-05 00:00:00},
	pages = {207-213},
	abstract = {<p>We describe the use of gel permeation chromatography (GPC) setup with four size exclusion columns for\&nbsp;analysis of enzymatically digested glycosaminoglycans (GAGs). This setup provides information about\&nbsp;the molecular weight (Mw) and concentration of all species (low and high Mw) present in the digests in a\&nbsp;single measurement. The data about the fraction with high Mw (often omitted in the analysis of GAG\&nbsp;digests) provide direct evidence about the mechanisms of action of the enzymes. We proved the\&nbsp;feasibility of this methodology by applying it to chondroitin sulfate (CS) substrates with different\&nbsp;molecular weight and sulfation pattern and using different enzymes (hyaluronidase and chondroitinase).\&nbsp;NMR analysis of the obtained digests fractionated by ultrafiltration confirmed the results obtained by\&nbsp;GPC setup and reveal further details about the degradation mechanisms: (i) both enzymes preferentially attack 4-sulfated chondroitin and (ii) additionally to the well documented endolytic activity of hyaluronidase we also observed a low lyase activity for this enzyme reflected in the detected minor exolytic breakage. Finally, we demonstrate that CS with medium molecular weight (12{\textendash}60 kDa) which is sulfated mainly at 6-position can be obtained in good yields by enzymatic digestion and following ultrafiltration.</p>},
	keywords = {chondroitin sulfate, Glycosaminoglycans, Hyaluronidase},
	doi = {10.1016/j.aca.2015.05.034},
	url = {http://ac.els-cdn.com/S0003267015007576/1-s2.0-S0003267015007576-main.pdf?_tid=3c6605e2-142a-11e5-a0b3-00000aacb360\&acdnat=1434460914_c372dbe283a7d23f6a6fa8f37e334d98},
	author = {Silva, C. and Novoa-Carballal, R. and Reis, R. L. and Pashkuleva, I.}
}
