Allo A-Sepharose 4B亲和层析分离糖缺失转铁蛋白(CDT)

Isolation of Carbohydrate-Deficient Transferring(CDT) by Allo-A Sepharose 4B Affinity Chromatography

目的建立Allo-A Sepharose亲和层析技术分离血清中糖缺失转铁蛋白（CDT）和转铁蛋白（Tf）。方法Allo-A与的偶联反应在0．1retool／LNaHCO3（含0．5mmol／L NaCl）内进行；Allo-A Sepharose4B亲和层析技术用于分离血清CDT和＇If；ELISA用于定量CDT和Tf0结果Allo-A Sepharose4B亲和层析图谱显示2个洗脱峰，CDT存在于第一洗脱峰，Tf存在于第二洗脱峰。ELISA测定结果显示，在5例正常血清中，CDT水平为4．53～4．76（CV=1．8～9．7％），Tf水平为255．3～524．9（CV=5．1～10．7％）；在5例患者血清中，CDT水平为3．04～4．20（CV：0．8～11．6％），Tf水平为6．05～34．73（CV=0．7～11．1％）。表明Allo-A Sepharose4B亲和层析技术在分离CDT和Tf中，具有较理想的可重复性。结论Allo—ASepharose4B亲和层析技术可用于CDT和Tf的分离，如果能够获得CDT参比血清，可进一步评价该技术的灵敏度。

Objective To establish Allo - A Sepharose 4B affinity chromatography for isolating CDT and Tf from serum. Methods Coupling reaction of Allo - A with CNBr - activated Sepharose 4B was conducted in 0. lmmol/L NaHCO3 containing 0. 5mmol/L NaC1. Allo - A Sepharose 4B affinity chromatography was used to isolate CDT and Tf from serum. Tf - ELISA was used to determine the levels of the isolated CDT and Tf. Results Elution profiles showed two major peaks. CDT was found in the first elution peak and Tf in the second peak. In the sera from 5 healthy volunteers, the level of CDT ranged 4. 53 ～4. 76（ CV = 1.8 ～9. 7% ）, and the level of Tf ranged 255.3 ～524. 9（ CV =5.1 ～ 10. 7% ） ;in the sera from 5 patients, the level of CDT ranged 3.04～ 4. 20 （ CV =0. 8 ～ 11.6% ）, and the level of Tf ranged 6. 05 ～34. 73 （ CV = 0. 7 ～ 11.1% ）. These findings indicated that Allo - A Sepharose 4B affinity chromatography exhibited a good reproducibility when used in isolation of CDT and Tf. Conclusion Allo - A Sepharose 4B affinity chromatography is a acceptable method for isolation of CDT and Tf. Its sensitivity remains to be evaluated, conditioned by the availability of a panel of CDT reference sera.