Purification of a highly modified RNA-aptamer. Effect of complete denaturation during chromatography on product recovery and specific activity.
J Chromatogr B Biomed Sci Appl
; 726(1-2): 237-47, 1999 Apr 16.
Article
de En
| MEDLINE
| ID: mdl-10348191
ABSTRACT
To evaluate RNA-aptamers as potential drug candidates, efficient and scaleable purification protocols are needed. Because aptamers are highly structured and rigid molecules, denaturation during the purification process is a critical aspect to obtain a pure and active product. A two-step chromatographic procedure was developed to purify a synthetic anti-VEGF aptamer at the preparative scale. A reversed-phase chromatographic step was optimized with a highly hydrophobic ion pairing reagent, followed by ion-exchange chromatography in which heat and a chaotropic salt were used. Because of the presence of 2'-modified ribose, denaturation conditions had to be optimized in both chromatographic steps to achieve a fully active molecule.
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Collection:
01-internacional
Base de données:
MEDLINE
Sujet principal:
ARN
/
Chromatographie en phase liquide à haute performance
/
Chromatographie d'échange d'ions
/
Dénaturation d'acide nucléique
Langue:
En
Journal:
J Chromatogr B Biomed Sci Appl
Sujet du journal:
QUIMICA CLINICA
Année:
1999
Type de document:
Article
Pays d'affiliation:
États-Unis d'Amérique