Method development for enrichment of autoantibodies from human plasma

University essay from KTH/Skolan för kemi, bioteknologi och hälsa (CBH)

Abstract: Antibodies are naturally occurring in humans, with the function to protect the body from pathogens. Occasionally, antibodies towards the body’s own proteins are produced. These so called autoantibodies are present in healthy individuals but are also highly associated with diseases with autoimmune involvement. Research on autoantibodies in healthy individuals as well as in patients is important to gain knowledge and facilitate prognostics, diagnostics and treatment. However, a method for purification of these antibodies has not previously been described. In the present project, an enrichment procedure of circulating autoantibodies found in human plasma is described. Twenty protein fragments previously known to be highly reactive were attached to magnetic microbeads, enabling autoantibodies from eight human plasma sample pools to be captured. The six antigens with highest shown reactivity were chosen for elution procedure. Using pH alterations and heat treatments, a successful elution and enrichment procedure was developed. With analysis of the eluted autoantibodies, it can be established that the enrichment was successful on multiple sample pools. In the scaled-up procedure, autoantibodies could be enriched in all positive antigen-sample combinations. Concentration measurements indicated amounts of up to 0.23 mg antibodies per ml eluate. This implies sufficient concentrations for further applications of the enriched autoantibodies.

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