Label-free differentiation and quantification of ricin, abrin from their agglutinin biotoxins by surface plasmon resonance

Abstract

Here we describe an affinity molecule-directed surface plasmon resonance (SPR) immunosensor for a label-free, differentiation and quantification of ricin and abrin from their structural highly like agglutinin biotoxins. By an introduction of protein G as the affinity capturing molecule, we fulfilled a complete strategy contains (i) screening monoclonal antibodies to be paired in a sandwiched format, (ii) differentiate quantification from the agglutinin, (iii) ascertain of active from inactive biotoxin, and (iv) structural identification of captured biotoxins on a single chip. By the aid of an enrichment step from immunomagnetic beads, we could accurately measure ricin or abrin with a concentration lowered to 0.6 ng/mL (10 pM) in different complex matrices such as stevia, protein powder, and human plasma, with linear ranges of two or three orders of magnitude, and satisfied recovery. We then differentially quantified the mixed crude extracts from castor beans and jequirity peas, and real samples from the fourth OPCW biotoxin exercise to prove the practical availability. We further provided a SPR-mass spectrometric evidence directly obtained from Protein G affinity chip via a noncovalent molecule surface for the first time for definitely structural identification for crude extracts.

Keywords: Abrin; Agglutinin; Differentiation; Mass spectrometry; Ricin; Surface plasmon resonance.