A Safe Tetramethylbenzidine Buffer for Enzyme Assays

描述（由申请人提供）：本提案的总体目标是 验证SEDUM的无害四甲基联苯胺水溶液（TMB） 用于辣根过氧化物酶测定的制剂。当前安全 与TMB的所有商业制剂相关的问题是它们都含有 有机溶剂最终的TMB产物是缓冲水溶液， 含有15- 20%的有机溶剂。使用的几种溶剂是 标记为有毒物质，如二甲基甲酰胺、二氧六环和甲醇。 一些供应商使用二甲基亚砜，但与危险的 溶剂后SEDUM开发了一种安全的TMB水性制剂， 稳定TMB，然后将其加入到水性缓冲液中。比较 使用这种新配方与现有的TMB产品进行的研究已经产生了 使用商业ELISA诊断产品作为对照获得类似的数据。数据 表明这种新配方将满足货架期要求 4 ℃下18个月的寿命要求。SEDUM现在建议验证这一点 过程TMB稳定化条件需要优化。适当的 缓冲区需要最终确定，这将产生一个产品，提供一个安全的 试剂，具有较高的测定灵敏度和最终的长期稳定性 制造产品。

DESCRIPTION (provided by applicant):The overall objective of this proposal is to validate SEDUM's non-hazardous, aqueous tetramethylbenzidine (TMB) formulation for horseradish peroxidase enzyme assays. One of the current safety issues associated with all commercial formulations of TMB is they all contain organic solvents. The final TMB product is a buffered aqueous solution that contains 15-20 percent organic solvent. Several of the solvents used are labeled as toxic substances such as, dimethylformamide, dioxane, and methanol. Some of the suppliers use dimethylsulfoxide but in combination with a hazardous solvent. SEDUM has developed a safe, aqueous formulation of TMB based on first stabilizing the TMB and then adding it to an aqueous buffer. Comparative studies with this new formulation with existing TMB products has produced similar data using commercial ELISA diagnostic products as controls. Data has been generated that indicates that this new formulation will satisfy the shelf life requirement of 18 months at 4 C. SEDUM now proposes to validate this process. The TMB stabilizing conditions need to be optimized. The appropriate buffer needs to be finalized that will produce a product that offers a safe reagent, has high assay sensitivity and the long-term stability of the final manufactured product.