ISSN:
0138-4988
Keywords:
Life Sciences
;
Life Sciences (general)
Source:
Wiley InterScience Backfile Collection 1832-2000
Topics:
Process Engineering, Biotechnology, Nutrition Technology
Notes:
Supernatants of a melanoma cell line were chromatographed on five affinity supports to evaluate and compare purification protocols for isolation of tissue-type plasminogen activator (t-Pa).t-PA was effectively adsorbed quantitatively from cell culture supernatants by lysine-Sepharose, concanavalin A-Sepharose, zinc-chelate-Sepharose and monoclonal antibody-Sepharose. The major part of t-PA was effectively adsorbed also by heparin-Sepharose. About 20 per cent of t-PA activity present in the culture medium however was not bound to the latter support, which indicates a heterogeneity of melanoma t-PA. The heterogeneity is not due to carbohydrate variants I and II since both vriants were adsorbed and eluted from all the affinity supports investigated.Highly enriched t-PA preparations were obtained by sequential chromatography on two supports. t-PA isolated by monoclonal antibody affinity chromatography and subsequently purified by gel filtration was of highest purity. Other useful combinations were chromatography on concanavalin A-Sepharose or lysine-Sepharose followed by chromatography on heparin-Sepharose, or zinc-chelate-Sepharose followed by chromatography on lysine-Sepharose.
Additional Material:
1 Tab.
Type of Medium:
Electronic Resource
URL:
http://dx.doi.org/10.1002/abio.370100409
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