Selective Adhesion of Antibodies on Patterned Polyelectrolytes within Microfluidic Networks

Darwin R. Reyes,a S. Patricia Becerra,b Laurie Locascio,c Michael Gaitana

aSemiconductor Electronics Division, Electronics and Electrical Engineering Laboratory, National Institute of Standards and Technology, Gaithersburg, MD

bNational Eye Institute, National Institutes of Health, Bethesda, MD

cAnalytical Chemistry Division, Chemical Science and Technology Laboratory, National Institute of Standards and Technology, Gaithersburg, MD

The micropatterning of antibodies in small and specific areas within microfluidic networks was performed. Polyelectrolyte multilayers (PEMs), consisting of alternating layers of polycations and polyanions adsorbed on a charged surface, were used to modify the functionality of discrete regions of poly(dimethylsiloxane) substrates. These altered regions provide a surface where antibodies can interact and ultimately anchor. Atomic force microscopy (AFM) characterization showed that the thickness of the patterned PEMs obtained with our method is within the range of the methods previously used. By using a constant flow system, fluorescent-labeled antibodies (Rabbit IgG and Goat Anti-Rabbit IgG) were transported throughout the channels and selectively anchored on the PEMs regions. The use of blocking agents, to avoid the adhesion of antibodies on areas not patterned, was not necessary. It was observed that the antibodies adhered better on poly(styrenesulfonic acid) surfaces when compared to poly(allylamine hydrochloride) and polyethylenimine surfaces. Antigen/antibody immunoassay type binding was attempted and binding between the complex antibody/antigen was observed. This patterning method required less than ten minutes to attach the antibodies to the surface.

 

 

Presenting Author’s information:

Darwin R. Reyes

Semiconductor Electronics Division

Electronics and Electrical Engineering Laboratory

Room B308, Technology Bldg. (225)

Mail Stop 8120

Tel.: (301) 975-5466

Fax: (301) 948-4081

darwin.reyes@nist.gov

Not a member of Sigma Xi

Category:

Biology and Biotechnology

Chemistry