Immunosensors for C-Reactive Protein Based on Ultrathin Films of Carboxylated Cellulose Nanofibrils.

C-reactive protein (CRP) is an acute phase protein that has been widely used as a predictor of cardiovascular diseases. We report herein the synthesis of immunosensors based on carboxylated cellulose nanofibrils (CNF) for CRP detection, as demonstrated by quartz crystal microgravimetry (QCM). QCM sensors carrying ultrathin films of carboxylated CNF were prepared by using two protocols: (i) spin coating of CNF on the sensors followed by carboxylation via in situ oxidation with 2,2,6,6-tetramethylpiperidine 1-oxyl and (ii) carboxymethylation of CNF in aqueous dispersion followed by spin coating deposition on the sensors. Protein A was conjugated to the carboxylated CNF via N-(3-(Dimethylamino)propyl)-N'-ethylcarbodiimide hydrochloride/N-hydroxysuccinimide and used as a ligand for oriented immobilization of anti C-reactive protein (anti-CRP). The different carboxyl group density of the two oxidized CNF surfaces influenced Protein A binding and, subsequently, the available immobilized anti-CRP molecules. The detection efficiency for CRP, specificity, and concentration range displayed by the carboxylated CNF-based immunosensors coupled with oriented and unoriented anti-CRP were determined and compared.