A High-Affinity Gold-Binding Camel Antibody: Antibody Engineering for One-Pot Functionalization of Gold Nanoparticles as Biointerface Molecules

Abstract

Antibodies, with their high affinity and specificity, are widely utilized in the field of protein engineering, medicinal chemistry, and nanotechnology applications, and our recent studies have demonstrated the recognition and binding of antibody for the surface on inorganic material. In this study, we generated a high-affinity gold-binding antibody fragment by a combination of peptide-grafting and phage-display techniques and showed the availability of the material-binding fragment for one-pot functionalization of nanoparticles as interface molecules. After a gold-binding peptide sequence was grafted into one of the complementarity determining regions of a single variable domain of a heavy-chain camel antibody, a combinatorial library approach raised by 20 times the affinity of the peptide-grafted fragment. The high-affinity gold-binding fragment (E32) spontaneously adsorbed on gold nanoparticles, and consequently the nanoparticles formed a stable dispersion in a high-ionic-strength solution. Multivalent and bispecific antibodies constructed on the E32 platform by means of fusion technology functionalized gold nanoparticles in one pot, and these functionalized nanoparticles could be used to obtain surface plasmon resonance scattering images of cancer cells and to spontaneously link two different nanomaterials. Here, we propose the bispecific antibodies as convenient interface molecules in the nanosized world.

Solid-binding peptides: smart tools for nanobiotechnology

“…Unfortunately, only a handful of antibodies that are able to recognise materials have been generated owing to their low immunogenicity [25]. A novel approach to the production of material-binding antibodies using SBPs has been established [26]. SBPs were grafted onto antibody fragments through genetic engineering to form a fusion protein, followed by phage display and selection.…”

“…The antibodies then were subjected to directed evolution to improve their binding avidity and specificity. Recently, these antibodies have been used to mediate the self-assembly of hybrid nanomaterials and the functionalisation of immunosensors [26,27]. Even though antibodies offer a structural scaffold for SBPs that can improve their stability and binding avidity, this approach is complex and suffers from a lack of versatility because it requires the specific modification of antibodies.…”

Peptide-functionalized iron oxide magnetic nanoparticle for gold mining

“…Among them, gold-binding peptides were the first group of peptides to be selected and characterized (So et al 2009a ; So et al 2009b ). Because of the intensive study in the gold-peptide interaction, particularly in terms of binding kinetics and free energy, researchers can understand, engineer, and control peptide-material interactions and exploit these to tailor novel materials and systems for practical applications (Hnilova et al 2008 ; Hattori et al 2012 ; Niide et al 2013 ; Seker et al 2014 ).…”

Site-selective orientated immobilization of antibodies and conjugates for immunodiagnostics development

“…[150,151,188] Hattori and colleagues reported the development of GBP fused antibodies to achieve one-step antibody immobilization. [189] Moreover, antibodies that can specifically bind to gold surface have been developed. [190,191] These antibodies were further constructed as bispecific fragments consisting of fragments of two different monoclonal antibodies that can recognize gold and the analyte proteins.…”