Cell-free measurements of brightness of fluorescently labeled antibodies.

Validation of imaging contrast agents, such as fluorescently labeled imaging antibodies, has been recognized as a critical challenge in clinical and preclinical studies. As the number of applications for imaging antibodies grows, these materials are increasingly being subjected to careful scrutiny. Antibody fluorescent brightness is one of the key parameters that is of critical importance. Direct measurements of the brightness with common spectroscopy methods are challenging, because the fluorescent properties of the imaging antibodies are highly sensitive to the methods of conjugation, degree of labeling, and contamination with free dyes. Traditional methods rely on cell-based assays that lack reproducibility and accuracy. In this manuscript, we present a novel and general approach for measuring the brightness using antibody-avid polystyrene beads and flow cytometry. As compared to a cell-based method, the described technique is rapid, quantitative, and highly reproducible. The proposed method requires less than ten microgram of sample and is applicable for optimizing synthetic conjugation procedures, testing commercial imaging antibodies, and performing high-throughput validation of conjugation procedures.

Temperature-dependent shape-responsive fluorescent nanospheres for image-guided drug delivery.

Temperature-responsive nanoparticles used in conjunction with hyperthermia promise to provide synergistic effects for increasing drug efficacy. We propose a near-infared (NIR) fluorescent system based on a upper critical solution temperature (UCST) polymer, ISP2, integrated with a NIR fluorescent dye HITC for in vivo tracking. The system forms a nanoparticle that increases its volume as temperature increases, similar to the expansion of a Hoberman sphere. The nanospheres nearly doubled in size, from 80 nm to 140 nm, during a temperature increase from 40°C to 60°C.