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《Chemical Research In Chinese Universities》 2000-12
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Coupling of DNA to Bead Type Cellulose as Immunoadsorbent

KONG De Ling *, DAI Jun, CHEN Chang Zhi, YU Yao Ting(Institute for Molecular Biology, Bioactive Materials Key Laboratory of Ministry of Education, Nankai University, Tianjin 300071, China) [WT5HZ]  
In order to remove the anti DNA antibody from the blood of patients with systematic lupus erythematosus(SLE), DNA immunoadsorbent was prepared by coupling calf thymus DNA to epichlorohydrin activated cellulose beads. The methods and conditions for activation of carrier beads, DNA coupling and adsorption with patient's plasma were investigated. The activation level was found to be dependent on the NaOH concentration, amount of offered epichlorohydrin and the molar ratio between NaOH and epichlorohydrin. One milliliter of cellulose beads was activated by incubation with 0.5 mL of epichlorohydrin and 2.0 mL of 2.0 ? ?.0 mol/L NaOH solution at 40 ℃ for 2 h, and the activation level was 60_100 μmol/mL beads. DNA coupling was found to be strongly dependent on temperature and pH. The optimal coupling conditions were 0.1 mol/L Tris HCl buffer, pH 7.9 ? ?.0 and 70—90 ℃. High activation level, high DNA concentration and long reaction time were also required. Under the optimal condition, 2.0 ? ?.0 mg of DNA could be coupled to 1.0 mL of cellulose beads. In vitro adsorption tests showed that incubation of 1.0 mL of adsorbent with 3.0 mL of plasma could remove 40%—70% of anti DNA antibody from the plasma. [WT5HZ]
【Fund】: 国家自然科学基金! (批准号 :2 96740 14 )资助
【CateGory Index】: O636;R318
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