| AVS 56th International Symposium & Exhibition | |
| Biomaterial Interfaces | Thursday Sessions |
| Session BI-ThP |
| Session: | Biomaterial Interfaces Poster Session II (Arrays, Sensing, Micro/Nanofabrication, SPM) |
| Presenter: | F. Liu, University of Utah |
| Authors: | F. Liu, University of Utah M. Dubey, University of Washington K. Emoto, Accelr8 Technology Corporation H. Takahashi, University of Utah D.W. Grainger, University of Utah D.G. Castner, University of Washington |
| Correspondent: | Click to Email |
Surface patterning is often used to immobilize bioactive molecules including proteins, oligonucleotides and small ligands, to localize surface reactions for bioassays and to provide desired cell and bacterial adhesion. This study reports extensive surface analysis of a commercial PEG-based surface chemistry with active ester (NHS)-activity in patterned films. The study followed sequential immobilization and masking reactions on photolithographic patterns used to immobilize peptides, proteins, and cultured cells to specific patterned regions of NHS-reactive or de-activated chemistry.[1] Biotin and peptide patterns were correlated to patterned reactive NHS surface chemistry using high-resolution time-of-flight secondary ion mass spectrometry (ToF-SIMS) for each species. Cell growth and patterning in 15-day serum cultures followed peptide patterns. In other patterned samples, mixed protein (streptavidin and HaloTag™) solutions produced spontaneous self-recognized, bound patterns on photolithographically surface-patterned affinity ligands for each (i.e., biotin and chloroalkane, respectively). The approach uses high-affinity protein-surface self-selection onto patterned PEG-NHS surfaces that exhibit intrinsically low non-specific adsorption background. Fluorescence images and ToF-SIMs imaging of the resulting protein surface selection from mixtures support highly specific interactions of proteins with their respective ligands patterned on the surface.[2] On-going work comparing ToF-SIMs imaging of antibody Fc and Fab fragments supports some ability to produce different whole antibody orientations on neighboring patterns spontaneously. Use of principal component analysis (PCA) helps to increase the ToF-SIMS image contrast and provide protein orientational details based on amino acid compositions.[3]
References:
[1]Takahashi, H., Emoto, K., Dubey, M., Castner, D. G., Grainger, D. W., Adv. Func. Matls. 2008. 18(14): p.2079-2088.
[2]Dubey M., Emoto K., Takahashi H., Castner D.G., Grainger, D.W., submitted, 2009.
[3] Dubey, M., Takahashi, H., Lew, F., Emoto, K., Castner, D.G., Grainger, D.W., submitted, 2009.