Retention studies of DNA on anion-exchange monolith chromatography Binding site and elution behavior.
Linear gradient elution experiments were carried out on monolithic anion-exchange chromatography (AEC) with oligo-DNAs of various sizes (4-50mer, molecular weight M(W)=1200-15,000) and compositions in order to investigate the retention mechanism. The binding site (B) values as well as the peak salt elution concentration I(R) values were determined. The B values determined for the monolithic AEC were similar to the values for non-porous AEC and porous AEC. The B value increased linearly with the number of charges (bases) of single-strand DNA when M(W) is less than ca. 3600 (12mer). When M(W) is greater than 6000, the slope of B versus M(W) decreased, and became very small at M(W)>30,000. The I(R) value also increased linearly with M(W) for M(W)<6000, and slightly with M(W) for M(W)>10,000. It was shown that a very difficult separation of a single-strand 50mer poly(T) and a double-strand 50mer poly(A) and poly(T) was accomplished within 10 min by using a very shallow gradient at a high initial salt concentration (0.5M) and a high flow-velocity (2.7 cm/min).
Yamamoto
Shuichi
S
Department of Chemical Engineering, School of Engineering, Applied Medical Engineering Science Division, Graduate School of Medicine, Yamaguchi University, Tokiwadai, Ube 755-8611, Japan. shuichi@yamaguchi-u.ac.jp
Nakamura
Masashi
M
Tarmann
Christina
C
Jungbauer
Alois
A
D000327:Adsorption; D000837:Anion Exchange Resins / Q000737:chemistry*; D001665:Binding Sites; D002852:Chromatography, Ion Exchange / Q000295:instrumentation* / Q000379:methods*; D004247:DNA / Q000737:chemistry / Q000302:isolation & purification*; D015203:Reproducibility of Results
D000837:Anion Exchange Resins; D004247:DNA