| Autor: |
Khan MA; Department of Chemical and Materials Engineering, University of Kentucky, Lexington, KY 40506-0046, USA., Ghanim RW; Department of Chemical and Materials Engineering, University of Kentucky, Lexington, KY 40506-0046, USA., Kiser MR; Department of Chemical and Materials Engineering, University of Kentucky, Lexington, KY 40506-0046, USA., Moradipour M; Department of Chemical and Materials Engineering, University of Kentucky, Lexington, KY 40506-0046, USA., Rogers DT; Naprogenix Inc., UK-ASTeCC, Lexington, KY 40506-0286, USA., Littleton JM; Naprogenix Inc., UK-ASTeCC, Lexington, KY 40506-0286, USA., Bradley LH; Department of Neuroscience, University of Kentucky, Lexington, KY 40536-0298, USA., Lynn BC; Department of Chemistry, University of Kentucky, Lexington, KY 40506-0055, USA., Rankin SE; Department of Chemical and Materials Engineering, University of Kentucky, Lexington, KY 40506-0046, USA., Knutson BL; Department of Chemical and Materials Engineering, University of Kentucky, Lexington, KY 40506-0046, USA. |
| Abstrakt: |
Successful strategies for the attachment of oligopeptides to mesoporous silica with pores large enough to load biomolecules should utilize the high surface area of pores to provide an accessible, protective environment. A two-step oligopeptide functionalization strategy is examined here using diazirine-based heterobifunctional linkers. Mesoporous silica nanoparticles (MSNPs) with average pore diameter of ~8 nm and surface area of ~730 m 2 /g were synthesized and amine-functionalized. Tetrapeptides Gly-Gly-Gly-Gly (GGGG) and Arg-Ser-Ser-Val (RSSV), and a peptide comprised of four copies of RSSV (4RSSV), were covalently attached via their N-terminus to the amine groups on the particle surface by a heterobifunctional linker, sulfo-succinimidyl 6-(4,4'-azipentanamido)hexanoate (sulfo-NHS-LC-diazirine, or SNLD). SNLD consists of an amine-reactive NHS ester group and UV-activable diazirine group, providing precise control over the sequence of attachment steps. Attachment efficiency of RSSV was measured using fluorescein isothiocyanate (FITC)-tagged RSSV (RSSV-FITC). TGA analysis shows similar efficiency (0.29, 0.31 and 0.26 mol peptide/mol amine, respectively) for 4G, RSSV and 4RSSV, suggesting a generalizable method of peptide conjugation. The technique developed here for the conjugation of peptides to MSNPs provides for their attachment in pores and can be translated to selective peptide-based separation and concentration of therapeutics from aqueous process and waste streams. |
