Serum protein electrophoresis (SPE) is widely used to quickly alert clinicians to a range of disorders. SPE separates serum proteins into characteristic bands that can reveal abnormalities such as monoclonal gammopathies, which are detected by analyzing the gamma peak of the SPE. Abnormal profiles in a SPE usually triggers a series of more specific immunoassays to confirm a diagnosis.<br/>Here we develop dual detection high-speed capillary electrophoresis (HSCE) for simultaneous SPE and immunoassay measurements. Backscatter interferometry (BSI) detects changes in refractive index, providing a universal signal for SPE measurements. The same excitation beam also excites fluorescence which is detected on a second channel. This enables SPE profiles and the detection of fluorescently tagged antigen-antibody complexes in the same detection volume, simultaneously. Separation speed is important since the antibody-antigen complex tends to dissociate during the separation process and the analysis must be performed before a significant portion of the complex is dissociated. The ultra-thin wall of the capillary enables high field strengths without Joule heating and the short capillary length leads to rapid separations.<br/>The dual detection HSCE approach is validated on serum samples spiked with fluorescein as an antigen. Samples incubated with an anti-fluorescein antibody are separated to generate simultaneous SPE and immunoassay profiles. Progress towards developing integrated analysis capabilities for the detection of disease will be presented.

Development of simultaneous refractive index and fluorescence detection platform using high-speed capillary electrophoresis

ACS Fall 2021

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