Orientation-Controlled Bioconjugation of Antibodies to Silver Nanoparticles
| Autor: | Leilani Smith, Nicole E. Pollok, Charlie Rabin, Richard M. Crooks |
|---|---|
| Rok vydání: | 2019 |
| Předmět: |
Silver
Biomedical Engineering Metal Nanoparticles Pharmaceutical Science Bioengineering 02 engineering and technology Orientation (graph theory) Conjugated system 01 natural sciences Antibodies Article Immunoglobulin G Silver nanoparticle Drug Stability Sulfhydryl Compounds Pharmacology Bioconjugation biology 010405 organic chemistry Chemistry Organic Chemistry 021001 nanoscience & nanotechnology Combinatorial chemistry 0104 chemical sciences Cross-Linking Reagents biology.protein Antibody 0210 nano-technology Biotechnology |
| Zdroj: | Bioconjugate Chemistry. 30:3078-3086 |
| ISSN: | 1520-4812 1043-1802 |
| DOI: | 10.1021/acs.bioconjchem.9b00737 |
| Popis: | Here we report on the use of heterobifunctional cross-linkers (HBCLs) to control the number, orientation, and activity of immunoglobulin G antibodies (Abs) conjugated to silver nanoparticles (AgNPs). A hydrazone conjugation method resulted in exclusive modification of the polysaccharide chains present on the fragment crystallizable region of the Abs, leaving the antigen-binding region accessible. Two HBCLs, each having a hydrazide terminal group, were synthesized and tested for effectiveness. The two HBCLs differed in two respects, however: (1) either a thiol or a dithiolane group was used for attachment to the AgNP; and (2) the spacer arm was either a PEG chain or an alkyl chain. Both cross-linkers immobilized 5 ± 1 Abs on the surface of each 20 nm-diameter AgNP. Electrochemical results, obtained using a half-metalloimmunoassay, proved that Abs conjugated to AgNPs via either of the two HBCLs were 4 times more active than those conjugated by the more common physisorption technique. This finding confirmed that the HBCLs exerted orientational control over the Abs. We also demonstrated that the AgNP-HBCL-Ab conjugates were stable and active for at least two weeks. Finally, we found that the stability of the HBCLs themselves was related to the nature of their spacer arms. Specifically, the results showed that the HBCL having the alkyl chain is chemically stable for at least 90 days, making it the preferred cross-linker for bioassays. |
| Databáze: | OpenAIRE |
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