Summary The adaptive immune system is thought to be a rich source of protein biomarkers, but diagnostically useful antibodies remain unknown for a large number of diseases. This is, in part, because the antigens that trigger an immune response in many diseases remain unknown. We present here a general and unbiased approach to the identification of diagnostically useful antibodies that avoids the requirement for antigen identification. This method involves the comparative screening of combinatorial libraries of unnatural, synthetic molecules against serum samples obtained from cases and controls. Molecules that retain far more IgG antibodies from the case samples than the controls are identified and subsequently tested as capture agents for diagnostically useful antibodies. The utility of this method is demonstrated using a mouse model for multiple sclerosis and via the identification of two candidate IgG biomarkers for Alzheimer's Disease.

Summary Several approaches have been developed for screening combinatorial libraries or collections of synthetic molecules for agonists or antagonists of protein function, each with its own advantages and limitations. In this report, we describe an experimental platform that seamlessly couples massively parallel bead-based screening of one bead one compound combinatorial libraries with microarray-based quantitative comparisons of the binding affinities of the many hits isolated from the bead library. Combined with other technical improvements, this technique allows the rapid identification of the best protein ligands in combinatorial libraries containing millions of compounds without the need for labor-intensive re-synthesis of the hits.

A rapid array-based protocol is presented by which a modest affinity protein-binding small molecule can be appended to a library of peptoids via Click chemistry. The array can then be screened for improved ligands that exhibit a higher affinity for the protein target.

A patient was found to have a positive direct antiglobulin test and thrombocytopenia while on a moderate dose of intravenous penicillin. Serological evaluation of the patient's red cells demonstrated that the positive antiglobulin test was due to antipenicillin antibody. This antibody also was demonstrated in the patient's serum. The patient's platelets had increased quantities of IgG; an eluate from her platelets gave positive test results with platelets treated with penicillin but not normal platelets. Her serum also reacted only with penicillin-treated platelets. Multiple absorptions of her serum with red cells treated with penicillin reduced reactivity against both fresh red cells and platelets treated with penicillin. This patient demonstrated the coexistence of drug-induced immune phenomena directed against both red cells and platelets.

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