Catalase-coupled gold nanoparticles: Comparison between the carbodiimide and biotin–streptavidin methods

Abstract

The use of proteins for therapeutic applications requires the protein to maintain sufficient activity for the period of in vivo treatment. Many proteins exhibit a short half-life in vivo and, thus, require delivery systems for them to be applied as therapeutics. The relative biocompatibility and the ability to form functionalized bioconjugates via simple chemistry make gold nanoparticles excellent candidates as protein delivery systems. Herein, two protocols for coupling proteins to gold nanoparticles were compared. In the first, the strong biomolecular binding between biotin and streptavidin was used to couple catalase to the surface of gold nanoparticles. In the second protocol, the formation of an amide bond between carboxylic acid coated gold nanoparticles and free surface amines of catalase using carbodiimide chemistry was performed. The stability and kinetics of the different steps involved in these protocols were studied using UV-Visible spectroscopy, dynamic light scattering, and transmission electron microscopy. The addition of mercaptoundecanoic acid in conjugation with (N-(6-(biotinamido)hexyl)-3′-(2′-pyridyldithio)-propionamide increased the stability of biotinylated gold nanoparticles. Although the carbodiimide chemistry based bioconjugation approach exhibited a decrease in catalase activity, the carbodiimide chemistry based bioconjugation approach resulted in more active catalase per gold nanoparticle compared to that of mercaptoundecanoic acid stabilized biotinylated gold nanoparticles. Both coupling protocols resulted in gold nanoparticles loaded with active catalase. Thus, these gold nanoparticle systems and coupling protocols represent promising methods for the application of gold nanoparticles for protein delivery.

Specific Enzyme Immobilization Approaches and Their Application with Nanomaterials

“…Chirra et al [25] made catalase biotinylation via succinimide chemistry. Au nanoparticles were first coated with biotin, and streptavidin was added as mediator for the conjugation of biotinylated catalase and biotin-coated Au nanoparticles, which can be used as delivery systems for therapeutic purpose.…”

A Simplified Method to Attach Antibodies on Liposomes by Biotin-Streptavidin Affinity for Rapid and Economical Screening of Targeted Liposomes

“…2 3 5-7 18-21 In addition, more recently the biotin STREP system was been investigated for use on other nanoparticle types, as for example for coupling of catalase on the surface of gold nanoparticles, 22 or for DNA-detection by magnetic nanoparticles. 23 Herein, a simple one-step biotin-STREP method for attachment of mAbs on LIPs was developed.…”

Shell-Nanostructured Materials for Biopharmacy and Biomedicine

“…With the advent of numerous tools, techniques, and concepts related to nanotechnology, in combination with the inherent property of gold to form functionalized bioconjugates via simple chemistry, gold has found importance in various biodiagnostic and therapeutic applications [25]. Herein, we detail the progress made in the functionalization of gold surfaces, both planar and particulates, at the nanoscale for diagnostic and therapeutic Surface modification of gold particles with stabilizing agents can be achieved by many methods.…”