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Quantitation of free chemiluminescent acridinium-9-carboxamide salts in bioconjugates John C. Gebler*; M Adamczyk; K Shreder; J Wu D-9NM AP-20, 100 Abbott Park Rd., Abbott Park, IL 60064-6016, USA      *(john.gebler@abbott.com) Acridinium-9-carboxamide salts are effective as chemiluminescent labeling reagents for use in immunoassays. During the conjugation process, the acridinium active ester undergoes a competitive hydrolysis to form the free acid, which may noncovalently bind to the protein or other assay components and thus, negatively assay performance. Therefore, the detection and quantitation of the residual label is an important aspect for the characterization of these chemiluminescent bioconjugates. A sensitive and robust liquid chromatography electrospray mass spectrometry method has been developed for the quantitative determination of noncovalently bound acridinium free acid in bioconjugates. The lower level of quantitation was 50 nM with a linear concentration range of 50-1000 nM. The method is specific, reproducible, and provides a new aspect for chemiluminescent conjugate characterization. [Poster: gebler.johnc.31792]

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