Unusual gold nanoparticle-antibody interactions

The formation of an antibody (Ab) protein corona surrounding gold nanoparticles (AuNPs) is a crucial step in the design of immunological assays. The Ab corona stabilizes AuNPs, preventing their aggregation even at high ionic strength, and can be achieved by simply mixing Abs and AuNPs. In this paper, we report the unusual interactions between AuNPs and the antibody against L1 Cell Adhesion Molecule (L1CAM) purified from rabbits. We have observed that at low ionic strength, the addition of a wide range of concentrations of rabbit monoclonal Abs against L1CAM protein immediately causes the coagulation of citrate-capped gold nanoparticles. This finding is surprising since the addition of proteins to colloidal gold usually forms a stable protein corona. The combination of extinction spectra, dynamic light scattering (DLS), and transmission electron microscopy (TEM) measurements reveals the presence of small clusters of AuNPs coated by the antibodies, as well as micron-sized antibody aggregates. Furthermore, static light scattering measurements demonstrate that Ab self-interactions are attractive (with a negative second virial coefficient, B2) and induce very slow Ab self-aggregation over several months. Overall, these results indicate that, at low ionic strength, the presence of AuNPs enhances Ab-Ab interactions, leading to their rapid aggregation. Simultaneously, the self-aggregation of the antibodies coating the AuNPs results in the formation of nanoparticle clusters. The addition of NaCl to increase the ionic strength fully reverses the coagulation of AuNPs (the Ab-coated AuNPs repel each other) and dissolves the Ab aggregates (the Ab interactions become repulsive, with a positive B2). The AuNPs-induced enhancement of the aggregation process can be explained by considering that the highly favorable binding of Abs on the gold surface compensates for the entropic penalty associated with Ab-Ab aggregation. The phenomenon we observed is specific to anti-L1CAM purified from rabbits and aligns with very old reports on AuNP coagulation induced specifically by the immunoglobulins present in the cerebrospinal fluid of patients suffering from neurosyphilis or multiple sclerosis (C. Lange Zeitschr. Chemotherap., 1912, 1, 44). It is reasonable to hypothesize that other antibodies exhibit this unusual behavior, so this work may aid in the interpretation of “anomalous” results that might otherwise be attributed to errors in fine-tuning AuNPs-Abs conjugation protocols.