Polymethacrylate Library Validation

     One of the project’s major goals is validation of the combinatorial and computational method using a library of 2000 polymethacrylates.  Combinatorial synthesis enables the production of immense polymer libraries and this project aims at testing the biological properties (protein attachment, cell attachment, and cell proliferation) of selected group of these polymers and then to use computational modeling to predict the biological properties of other polymers in the library. Following computational based predictions, biological experiments are completed to validate or disprove the modeling methods.  

     In this assay each polymer is spincoated on the surface of the glass coverslip and then seeded with NIH3T3 cells. After a four-hour incubation, the cell adherence is examined using Aqueous One (Promega) reaction, which is based on the principle of adsorbed cells reacting with the reagent and producing a signal at λ=490. Greater cell attachment leads to higher level of signal measured. The proliferation assay is also based on this principle, except that the cells are incubated for additional four days, after which the reaction is performed and optical density (OD) signal is read.  Protein attachment assays are conducted by adsorption of fibrinogen onto the surface of solvent-cast materials and quantified by immunodetection with a fluorescently labeled antibody directed against fibrinogen.

Project Leader: Jared Bushman, PhD
Funding Source: RESBIO - The National Resource for Polymeric Biomaterials via the National Institutes of Health                       (NIH grant EB001046)