ISOELECTRIC FOCUSING PROCEDURES FOR ALLERGENIC EXTRACTS

等电聚焦已被证明是一种有用的工具 过敏性提取物。这种类型的电泳法可作为 用免疫印迹法检测提取物的免疫活性 过程，该过程被动地传输聚焦摘录并标识 产品中的IGE绑定组件。这两个配套的方法有 多种用途。提交标准化的提取物的身份可以 在免疫球蛋白E结合方面与参考提取物进行检查和比较 活性以及染色的IEF模式。最佳提取方法， 在产生的IgE结合蛋白方面，可以用这些来确定 有两种方法。过敏性疾病患者血清中IgE结合的差异 可对个人进行调查，并可选择血清用于测试池 使用这些方法，从而确保IgE结合的公平表示 在定量测试中的活跃性。另一个改编，使用SELECT 在免疫印迹中抑制IgE结合的成分，有助于 交叉反应性IgE结合组分的鉴定。准备中的IEF， 使用一种新的仪器，允许分离单个组件 从过敏原提取物中分离非IgE结合和IgE结合 提供足够数量的部件，以供分析和进一步测试。 在IEF之后，这些成分被进行二维电泳 以作进一步检查。IEF和免疫印迹已经成为一个关键 过敏性提取物稳定性研究评价中的因素。 用IEF作为初步筛查方法，丢失明确的IEF 条带模式通常表示某些过敏原活性丧失。 在对IEF进行评估之后，可以进行定量测试以确认 过敏原活性丧失。IEF也已被证明在 关于演示的标准化猫提取物的评价 批次之间的一致性，以及与以下相关的成分差异 致敏源材料。有人提议，IEF将成为一个 与FELD一起使用的标准化CAT提取物的发布标准 I分析和蛋白质分析已经是必需的。

Isoelectric Focusing has proven to be a useful tool in the evaluation of allergenic extracts. This type of electrophoresis serves as a basis for examining the immunological activity of the extract, using a blotting procedure that passively transfers the focused extracts and identifies the IgE binding components in the products. The two companion methods have multiple uses. The identity of extracts submitted for standardization can be examined and compared to reference extracts with respect to IgE binding activity as well as stained IEF patterns. The optimal extraction method, in terms of IgE binding proteins generated, may be determined with these two methods. Differences in IgE binding in the sera of allergic individuals may be surveyed, and sera may be selected for testing pools with these methods, thereby assuring a fair representation of IgE binding activity in quantitative tests. A further adaptation, using select components to inhibit IgE binding in the immunoblotting, aids in the identification of cross reactive IgE binding components. Preparative IEF, using a new apparatus, has permitted the isolation of single components from allergenic extracts, separating non-IgE binding and IgE binding components in sufficient quantities for analysis and further testing. Following IEF, these components are being subjected to 2-D electrophoresis for further examination. IEF and immunoblotting has become a critical factor in the evaluation of stability studies for allergenic extracts. Using IEF as the preliminary screening method, the loss of well defined IEF banding patterns frequently indicates some loss of allergenic activity. Following evaluation of IEF, quantitative tests can be performed to confirm loss of allergenic activity. IEF has also proven to be useful in the evaluation of standardized cat extracts, with respect to demonstrating consistency between lots, and compositional differences related to allergenic source material. It has been proposed that IEF will become a release criteria for standardized cat extracts, in conjunction with Fel d I assays and protein assays that are already required.