Rahman H, Skillen A W, Channon S M, Ward M K, Kerr D N
Clin Chem. 1985 Dec;31(12):1969-73.
We describe methods for studying the binding of Al by protein in serum: ultrafiltration, gel filtration, and immuno-affinity chromatography. For ultrafiltration we used an Amicon YM10 cellophane membrane with a nominal cutoff of 10 000 Da to separate ultrafiltrable and non-ultrafiltrable Al. For gel filtration we used Sephacryl S-300, and for immuno-affinity chromatography we used anti-transferrin coupled to CNBr-activated Sepharose to identify the Al-binding protein. For 30 normal subjects 54% of the total Al in serum was non-ultrafiltrable; for 30 patients with chronic renal failure being treated by hemodialysis 67% was non-ultrafiltrable. In both groups transferrin was identified as the major Al-binding protein in the serum. Results of gel-filtration studies should be interpreted with caution: some gel media adsorb "free" Al, which can be subsequently taken up by transferrin or desferrioxamine passing through the column. We find affinity chromatography to be a specific and reliable method, suitable for use in quantitative studies.
超滤法、凝胶过滤法和免疫亲和色谱法。超滤时，我们使用标称截留分子量为10000道尔顿的Amicon YM10玻璃纸膜来分离可超滤和不可超滤的铝。凝胶过滤用的是Sephacryl S - 300，免疫亲和色谱则使用偶联到溴化氰活化琼脂糖上的抗转铁蛋白来鉴定铝结合蛋白。对于30名正常受试者，血清中总铝的54%不可超滤；对于30名接受血液透析治疗的慢性肾衰竭患者，67%不可超滤。在两组中，转铁蛋白均被鉴定为血清中的主要铝结合蛋白。凝胶过滤研究结果的解释应谨慎：一些凝胶介质会吸附“游离”铝，随后这些铝可能会被通过柱子的转铁蛋白或去铁胺摄取。我们发现亲和色谱是一种特异且可靠的方法，适用于定量研究。
