Obtaining an immunoaffinity monolithic material: poly(GMA-co-EDMA) functionalized with an HPV-derived peptide using a thiol–maleimide reaction

Abstract

Metabolites have great potential for the design of biomarkers, since their presence or absence provides valuable information about a biological system. In this context, polyclonal antibodies are important metabolites for diagnostic procedures, but in some pathologies, it has been found that these metabolites are present at low concentrations, so it could be difficult to detect them. In this investigation, an organic monolithic material of poly(GMA-co-EDMA) was functionalized with a peptide via Michael addition (thiol–maleimide) click chemistry. The peptide, covalently bound to the monolith, contains the SPINNTKPHEAR sequence derived from the human papilloma virus L1 protein. It was determined that the obtained monolithic support allows selectively isolating polyclonal antibodies against the SPINNTKPHEAR sequence, since they are retained on the chemical surface of the material by an immunoaffinity interaction. The monolithic material functionalization protocol reported here could be applied to incorporate any peptide with a terminal cysteine in order to recover a specific analyte. A new method was developed for isolating and pre-concentrating antibodies using monolithic materials, which could contribute to the improvement of disease detection strategies based on immunoaffinity interactions.