Autoreactive antibodies raised by self derived de novo peptides can identify unrelated antigens on protein microarrays. Are autoantibodies really autoantibodies?

The development of arrays of human proteins has been a huge boon to the search for autoantibody diagnostics. Typically, slides with thousands of recombinant human proteins arrayed in an addressable fashion are incubated with sera from diseased or normal people. If an antibody binds a protein more in the diseased than in the normal cohort it is considered an autoantibody response. It is usually presumed that the autoantibody was elicited by the protein bound on the array. However, our studies using human protein and random peptide arrays indicate that antibody specificity may not be as high as commonly thought. Therefore we have tested the assumption of the source of autoantibodies. One test was to generate antibodies to two totally random peptides and bind these antibodies to a human protein array. One of the antibodies generated bound two human proteins. A second test was to generate an antibody to a frameshift peptide occurring in cancers. This antibody also bound several proteins on the array. We conclude that one should be cautious about assuming a particular autoantibody target on an array which elicited the original immune response.

► Does serum reactivity on proteome arrays correspond to the antigen?
► Proteome arrays were probed with sera raised against random and frameshift peptides.
► ELISA confirmed random peptides bound proteins identified on the array.
► ELISA confirmed a frameshift peptide bound a protein identified on the array.
► Multiple information sources are needed to associate immunogens and autoantibodies.