空气样品洗脱液的测定。

Anne Renström, Susan Gordon
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引用次数: 2

摘要

经过空气取样和洗脱后,空气样品洗脱液与其他物质一起含有未知数量的过敏原。感兴趣的蛋白质可以用免疫测定法定量,这是敏感的,经济的,并且可以用于高通量。然而,空气样本中抗原或过敏原的含量可能很低,因此测定必须非常敏感和特异性。免疫测定法使用抗体来捕获和可视化所选抗原。高特异性和敏感性可以最好地通过使用纯化,表征和特异性抗体来实现。可以在各种各样的测定装置和试剂之间进行选择。本文所描述的方法是为测量空气传播的啮齿动物过敏原而开发的。它是一种利用多克隆抗体的非竞争性、双位点(三明治)EIA。检测系统采用生物素和链亲和素提高灵敏度,以辣根过氧化物酶为底物,以3,3',5,5'-四甲基联苯胺(TMB)快速显色,灵敏度高。
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Assay of air sample eluates.

After air sampling and elution, the air sample eluate contains an unknown amount of allergens together with other materials. The proteins of interest can be quantified using immunoassays, which are sensitive, economical, and can be used for high throughput. However, the amount of antigen or allergen in an air sample may be very low and consequently the assays must be very sensitive and specific. Immunoassays use antibodies both to capture and visualize the chosen antigen. High specificity and sensitivity can best be achieved by the use of purified, characterized, and specific antibodies. It is possible to choose between a wide variety of assay setups and reagents. The method described here has been developed for the measurement of airborne rodent allergens. It is a noncompetitive, two-site (sandwich) EIA that utilizes polyclonal antibodies. The detection system uses biotin and streptavidin for increased sensitivity and horseradish peroxidase as the substrate with 3,3',5,5'-tetramethylbenzidine (TMB) for rapid color development and high sensitivity.

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