Cruschelli L, Clerico A, Penno G, Navalesi R, Giampietro O
Cattedra di Malattie del Metabolismo, Università degli Studi di Pisa, Italy.
Acta Diabetol Lat. 1990 Oct-Dec;27(4):349-56. doi: 10.1007/BF02580940.
Affinity chromatography by m-aminophenylboronic acid has been proposed for routine measurement of glycated albumin. We assayed glycated and non-glycated fractions of serum albumin (HSA) eluted by affinity chromatography columns by both a specific RIA method for the human serum albumin (HSA) and by a colorimetric method. Sixteen diabetic patients presented a significantly higher percentage of glycated-HSA than 7 control subjects with both methods, and a strong correlation was found between the values obtained with the two methods. RIA was able to detect a significant concentration of glycated-HSA in all normal subjects, while the colorimetric method was not. The accuracy of separation between the glycated and non-glycated fractions of albumin was tested using [14C]glucose as tracer. When [14C]glycated-HSA purified by Sephadex G25 filtration was chromatographed using the m-aminophenylboronic acid, only 5.3% of the total 14C-radioactivity present in the solution was recovered in the bound fraction, while 44.0% was eluted in non-protein-bound fraction and 54.7% was retained in the column. Our findings confirm that affinity chromatography by m-aminophenylboronic acid can be a useful tool in the monitoring of short glycemic control of diabetic patients. Our data also indicate that the affinity chromatography with m-aminophenylboronic acid does not accurately discriminate between glycated and non-glycated fraction of HSA.
有人提出用间氨基苯硼酸亲和色谱法来常规测定糖化白蛋白。我们通过一种针对人血清白蛋白（HSA）的特异性放射免疫分析法（RIA）和比色法，对亲和色谱柱洗脱的血清白蛋白（HSA）的糖化和非糖化部分进行了测定。16例糖尿病患者的糖化HSA百分比在两种方法下均显著高于7例对照受试者，且两种方法所得值之间存在强相关性。RIA能够检测到所有正常受试者中糖化HSA的显著浓度，而比色法不能。以[¹⁴C]葡萄糖为示踪剂，测试了白蛋白糖化和非糖化部分之间分离的准确性。当用Sephadex G25过滤纯化的[¹⁴C]糖化HSA通过间氨基苯硼酸进行色谱分析时，溶液中存在的总¹⁴C放射性中只有5.3%在结合部分中回收，而44.0%在非蛋白结合部分中洗脱，54.7%保留在柱中。我们的研究结果证实，间氨基苯硼酸亲和色谱法可作为监测糖尿病患者短期血糖控制的有用工具。我们的数据还表明，间氨基苯硼酸亲和色谱法不能准确区分HSA的糖化和非糖化部分。
