Analysis of the KHRI-3 Antigen in Guinea Pig Inner Ear and Lung Tissues
Braunz, Andrea E.
Autoimmune Sensorineural Hearing Loss (AISNHL) is a quickly progressing hearing loss that results from an immune response in one's body. In an effort to further understand and more effectively treat AISNHL, a monoclonal antibody (KHRI-3) and a polyclonal antibody (rabbit anti-CTL2) were developed, using the antigen found in AISNHL patients, for research using guinea pigs as a model. When the KHRI-3 antigen is probed with the KHRI-3 or the rabbit anti-CTL2 antibody after immunoprecipitation and SDS-PAGE, a doublet of approximately 68 kDa and 72 kDa is seen in Western Blots. Probing homogenates of guinea pig organs (brain, tongue, colon, and lung) with either antibody resulted in a single band with a molecular weight of -70 kDa To investigate why the inner ear extracts appear as a doublet, we performed several experiments dealing with the glycosylation on the KHRI-3 antigen. We hypothesized that sialic acid residues accounted for the upper band of the doublet in the inner ear antigen. However, the experiments demonstrated that sialic acid residues are found on the KHRI-3 antigen collected from the inner ear as well as the lung, revealing that the upper band in the inner ear is not due to sialic acid residues. Further research is needed to determine why the inner ear extracts appear as a doublet.
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