Not all protein-mediated single-wall carbon nanotube dispersions are equally bioactive

Abstract

Single-wall carbon nanotubes (SWCNTs) have been dispersed with proteins to increase biocompatibility and specificity, but examinations of dispersion parameters on functional cellular uptake are required for utilization of SWCNTs in biological applications. Here we correlate conditions of SWCNT dispersion with various proteins to uptake these SWCNTs in NIH-3T3 fibroblasts and J774A.1 macrophage-like cells. We varied protein types (bovine serum albumin – BSA, lysozyme – LSZ, and γ-globulins – γG), protein : SWCNT ratio and sonication time. Each protein created stable, high yield (∼25%) dispersions in water while preserving intrinsic SWCNT fluorescence, but SWCNT–LSZ flocculated in media and SWCNT–γG formed clusters in both water and media, drastically altering cellular internalization. Dispersion quality and yield improved with increased protein : SWCNT – without substantial effects from depletion attraction, even at 100 : 1 protein : SWCNT – and slightly increased internalized SWCNTs for both NIH-3T3 and J774A.1 cells. Longer sonication time (12 versus 2 h) improved the dispersion yield and quality but caused minor damage to SWCNTs and altered protein structure. Cell association of SWCNT–BSA was homogenous and unaltered by sonication time. Bulk assay showed that cell association of SWCNT–LSZ and SWCNT–γG was altered with 12 versus 2 h sonication, but imaging of individual cells showed that these differences are likely from precipitation of clusters of SWCNT–LSZ and SWCNT–γG in media onto cells. Hence, the quality of SWCNT–protein dispersions in water does not necessarily correlate with bulk cellular uptake, and quantification at the level of individual cells is required to determine delivery efficacy.