| This dissertation describes the development of covalent and noncovalent labeling schemes for the study of bioaffinity interactions via capillary electrophoresis with near infrared laser induced fluorescence detection. Specifically, noncovalent labeling schemes are used a novel method for the determination of drug binding constants via a near infrared dye displacement assay. Covalent labeling schemes are used for capillary electrophoretic immunoassays with near infrared laser induced fluorescence detection.; Typical binding studies done by capillary electrophoresis involve multiple runs to arrive at a binding constant. Relatively large amounts of reagent are consumed. Additionally, if visible laser induced fluorescence detection is used, sample autofluorescence is an issue. For these reasons, a competitive dye displacement assay for the determination of drug binding constants toward subdomain IIIA of human serum albumin was developed. Noncovalent labeling schemes are used for the assay. The assay allows for determination of binding constants in a single run. Additionally, the assay is fast, sensitive, and amenable to full automation.; Immunoassays are analytical techniques used in a wide variety of fields including environmental, clinical, and pharmaceutical chemistry. Immunoassays rely on the highly specific interaction between an antibody and antigen. Despite their wide use, immunoassays are not without their disadvantages. The technique is tedious and suffers from poor reproducibility. For these reasons, a capillary electrophoresis based immunoassay was developed. Two systems were studied; insulin and human serum albumin. The near infrared dye NN382 was covalently attached to both of the molecules. The capillary electrophoresis based immunoassay combines the high separation efficiency of capillary electrophoresis, the high specificity of immunoassays, and the high sensitivity of near infrared laser induced fluorescence detection. |
