Quantification of the Binding Affinity of a Specific Hydroxyapatite Binding Peptide

Published

January 27, 2010

Author(s)

Michael C. Weiger, Jung Jin Park, Marc D. Roy, Christopher M. Stafford, Alamgir Karim, Matthew L. Becker

Abstract

The genesis of bone and teeth results from highly coordinated processes which involve several cell types, proteins which direct the nucleation and crystallization of inorganic hydroxyapatite (HA). Recent studies have shown that engineered peptide sequences can either promote the nucleation process, control HA microstructure or even inhibit HA mineralization. Using phage display technology, a short peptide was identified that binds to crystalline HA and to HA-containing domains of human teeth with chemical and morphological specificity. However, the binding affinity and specific amino acids that significantly contribute to this interaction require further investigation. In this study, we employ a microfluidic chip based surface plasmon resonance imaging (SPRi) technique to quantitatively measure peptide affinity. We show the 4 layer fabrication and characterization of a novel HA SPR sensor supported on a glass substrate. We find the peptide (SVSVGMKPSPRPGGGK) binds with relatively high affinity (KD = 14.1 μM ± 3.8) to HA. The independently measured amino acid fragment SVSV seems to impart a significant contribution to this interaction while the MKPSP fragment may provide a conformational dependent component that enhances the peptides affinity but by itself shows little specificity in the current context. These data show that together, the two moieties promote a stronger synergistic binding interactions to HA than the simple combination of the individual components.

Citation

Biomaterials

Volume

31

Issue

11

Pub Type

Journals

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Keywords

Surface plasmon resonance imaging, hydroxyapatite, microfluidics, peptides

Citation

Weiger, M. , , J. , Roy, M. , Stafford, C. , Karim, A. and Becker, M. (2010), Quantification of the Binding Affinity of a Specific Hydroxyapatite Binding Peptide, Biomaterials, [online], https://tsapps.nist.gov/publication/get_pdf.cfm?pub_id=903862 (Accessed October 15, 2024)