(570f) Capture and Concentration of Pathogens in Chaotic Flows

Authors: 
Trujillo-de Santiago, G., Tecnológico de Monterrey
Prakash, G., Harvard-MIT
Risso, A., Brigham and Women's Hospital, Harvard Medical School
del Toro Runzer, C., Harvard-MIT
Hernández Medina, R., Centro de Biotecnología-FEMSA, Tecnológico de Monterrey, Escuela de Ingeniería y Ciencias
Alvarez, M. M., Centro de Biotecnología-FEMSA, Tecnológico de Monterrey, Escuela de Ingeniería y Ciencias
Khademhosseini, A., Massachusetts Institute of Technology
Infectious diseases of both viral and bacterial origin continue to be a health threat to millions of people in developed and underdeveloped countries. One successful treatment strategy for cases of sepsis or viral infections, such as Ebola disease and HIV, is the effective capture and removal of the pathogenic agent from the bloodstream (i.e., pathogen blood cleansing). We are presently developing filter-less technologies for the direct capture of bacteria (i.e., Escherichia coli) and eventually viruses (i.e., Ebola virus-like-particles) from the bloodstream. We use a portable/disposable system for the continuous capture of pathogens circulating through a microfluidic chamber. The system is based on the specific recognition of proteins on E. coli membranes; it integrates the use of (a) anti-E. coli polyclonal antibodies, (b) magnetic nanoparticles (MNP), (c) a microfluidic chaotic flow system, and (d) a neodymium magnet. Anti-E. coli antibodies are covalently immobilized within commercial magnetic nanoparticles to fabricate nanoparticles that will bind E. coli bacteria. Our experiments compare the performance of different immobilization strategies (amino-carboxylic covalent binding and streptavidin-biotin binding) and different magnetic nanoparticle sizes (range 30â??800 nm). The heart and distinctive feature of our system is a microfluidic chamber in which the E. coli binding particles and the bacteria are mixed by the action of a laminar chaotic flow produced by the alternating rotation of two cylinders. The intimate contact induced by this chaotic laminar flow promotes the capture of the bacteria by individual nanoparticles or nanoparticle clusters. The trapped E. coli are then concentrated by a simple magnet located downstream from the microchamber.

 This platform has key advantages over currently available methods, which are mostly based on the use of microfluidic channels or filtering membranes: (a) It is faster (overall capture times in the order of 1â??5 minutes); (b) It offers superior capture due to the intimate mixing induced by the chaotic flow; (c) It is easy to use, which reduces labor efforts and eliminates the need for dedicated and costly infrastructure.