Abstract
The large scale patterning of therapeutic proteins is a key to the efficient design, characterization, and production of biologics for cost effective, high throughput, and point-of-care detection and analysis system. We demonstrate an efficient method for protein deposition and adsorption on nanoporous silica substrates in specific patterns using a method called “micro-contact printing”. Multiple color-tagged proteins can be printed through sequential application of such micro-patterning technique. Two groups of experiments were performed. In the first group, the protein stamp was aligned precisely with the printing sites, where the stamp was applied multiple times. Optimal conditions were identified for protein transfer and adsorption using the pore size of 4 nm and thickness of 30 nm porous silica thin film. In the second group, we demonstrate the patterning of two-color rabbit immunoglobin labeled with fluorescein isothiocyanate and tetramethyl rhodamine iso-thiocyanate on porous silica substrates that have a pore size 4 nm, porosity 57% and thickness of the porous layer 30 nm. A pair of protein stamps, with corresponding alignment markings and coupled patterns, were aligned and used to produce a two-colored stamp pattern of proteins on porous silica. Different colored proteins can be applied to exemplify the diverse protein composition within a sample. This method of multicolor microcontact printing can be used to perform a fluorescence-based patterned enzyme-linked immunosorbent assay to detect the presence of various proteins within a sample.
Highlights
Microcontact printing is a method in which a stamp is ‘‘inked’’ with specific molecules with subsequent deposition of those molecules onto a substrate with wellcontrolled pattern
We demonstrate the patterning of two-color rabbit immunoglobin labeled with fluorescein isothiocyanate and tetramethyl rhodamine iso-thiocyanate on porous silica substrates that have a pore size 4 nm, porosity 57% and thickness of the porous layer 30 nm
We demonstrate the application of multicolor lCP for multiple antigen detection in immunoassays by performing a sandwich Enzyme-linked immunoabsorbent assay (ELISA) for the simultaneous detection of two common allergens, ovomucoid found in egg white, and casein found in milk
Summary
Microcontact printing (lCP) is a method in which a stamp is ‘‘inked’’ with specific molecules with subsequent deposition of those molecules onto a substrate with wellcontrolled pattern. There are many different types of ELISA, the sandwich ELISA technique described here will be most applicable to the investigation It involves affixing an amount of antigen to a surface via a capture antibody and washing the surface with a specific primary antibody. We demonstrate an efficient method for the deposition and adsorption of two different color-tagged proteins on a single nanoporous silica substrate through sequential application of the lCP technique. We demonstrate the application of multicolor lCP for multiple antigen detection in immunoassays by performing a sandwich ELISA for the simultaneous detection of two common allergens, ovomucoid found in egg white, and casein found in milk. The ability to miniaturize and microscale the immunoassay itself enables wider, more cost efficient, varieties of biomedical point of care applications
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
