Myint T, Hoshi S, Ookawara T, Miyazawa N, Suzuki K, Taniguchi N
Department of Biochemistry, Osaka University Medical School, Japan.
Biochim Biophys Acta. 1995 Oct 17;1272(2):73-9. doi: 10.1016/0925-4439(95)00067-e.
A polyclonal antibody specific for the Amadori compound, a product of an early stage of the Maillard reaction, was raised in rabbits by immunization with hexitol-lysine (1-glucitol-lysine or 1-mannitol-lysine) coupled with various carrier proteins. The affinity purified antibody has a high titre and preferentially recognizes the glucose adduct, in the presence of sodium borohydride, as judged on enzyme-linked immunosorbent assay as well as immunoblot analysis. The glycated proteins (Amadori products) in various tissues of normal and streptozotocin-induced diabetic rats were examined by immunoblot analysis. In diabetic conditions, kidney, liver, lens, brain and lung proteins are more susceptible to glycation than other tissue proteins. Heart, spleen, adrenal gland and muscle proteins exhibit similar extents of glycation in both normal and diabetic conditions. This is the first demonstration of a specific antibody against the Amadori compound being raised with a synthetic compound, and of the tissue distribution of glycated proteins in normal and diabetic conditions. The antibody was very useful for in vitro and in vivo experiments on the Maillard reaction.
通过用与各种载体蛋白偶联的己糖醇赖氨酸（1-葡糖醇赖氨酸或1-甘露糖醇赖氨酸）免疫家兔，制备了一种针对美拉德反应早期产物阿马多里化合物的多克隆抗体。经亲和纯化的抗体具有高滴度，在硼氢化钠存在下，通过酶联免疫吸附测定和免疫印迹分析判断，该抗体优先识别葡萄糖加合物。通过免疫印迹分析检测了正常大鼠和链脲佐菌素诱导的糖尿病大鼠各种组织中的糖化蛋白（阿马多里产物）。在糖尿病状态下，肾脏、肝脏、晶状体、大脑和肺组织中的蛋白质比其他组织蛋白更容易发生糖化。在正常和糖尿病状态下，心脏、脾脏、肾上腺和肌肉蛋白的糖化程度相似。这是首次证明用合成化合物制备出针对阿马多里化合物的特异性抗体，以及首次证明正常和糖尿病状态下糖化蛋白的组织分布情况。该抗体对美拉德反应的体外和体内实验非常有用。
