Reynoso G, Tuggey R, Hansen H, Fontelo P A, Konopka S, Miller J T
Am J Clin Pathol. 1981 Jun;75(6):786-93. doi: 10.1093/ajcp/75.6.786.
A ligand assay specific for cobalamin that uses mouse stomach as the source of intrinsic factor has been developed. When mouse stomach extract incubated with radiocobalamin is fractionated by gel chromatography, the radioactive complex elutes as a single peak with apparent molecular weight of 54,900. Formation of the complex is greater than 98% inhibited by human anti-intrinsic factor antibody. When the equivalent of 10,000 pg/ml of cobinamide is added to serum, the apparent cobalamin concentration detected averages 8.5 pg/ml. Correlation with the Lactobacillus leichmannii microbiologic assay results in the regression equation y = 0.97x + 20. In six patients who had megaloblastic anemia the serum cobalamin by the mouse intrinsic factor ligand assay ranged from 0 to 9 pg/ml. Because the primary source of intrinsic factor is free of R proteins, there is no need for extensive purification of the extract. The assay is sensitive, precise, and accurate, and no more difficult to perform than other conventional ligand assay procedures.
已开发出一种针对钴胺素的配体分析方法，该方法使用小鼠胃作为内因子的来源。当用放射性钴胺素孵育的小鼠胃提取物通过凝胶色谱法进行分离时，放射性复合物以单一峰的形式洗脱，表观分子量为54,900。人抗内因子抗体可抑制该复合物形成的98%以上。当向血清中添加相当于10,000 pg/ml的钴胺酰胺时，检测到的表观钴胺素浓度平均为8.5 pg/ml。与赖氏乳杆菌微生物学分析结果的相关性得出回归方程y = 0.97x + 20。在6例巨幼细胞贫血患者中，通过小鼠内因子配体分析测得的血清钴胺素浓度范围为0至9 pg/ml。由于内因子的主要来源不含R蛋白，因此无需对提取物进行广泛纯化。该分析方法灵敏、精确且准确，操作难度并不比其他传统配体分析方法更大。
