دورية أكاديمية
Immunoassays for Extracellular Vesicle Detection via Transmembrane Proteins Using Surface Plasmon Resonance Biosensors.
| العنوان: | Immunoassays for Extracellular Vesicle Detection via Transmembrane Proteins Using Surface Plasmon Resonance Biosensors. |
|---|---|
| المؤلفون: | Lopez Baltazar JM; Robert Frederick Smith School of Chemical and Biomolecular Engineering, Cornell University, Ithaca, New York 14853, United States., Gu W; Meinig School of Biomedical Engineering, Cornell University, Ithaca, New York 14853, United States., Bocková M; Robert Frederick Smith School of Chemical and Biomolecular Engineering, Cornell University, Ithaca, New York 14853, United States.; Institute of Photonics and Electronics, AS CR Chaberská57, Prague 182 51, Czech Republic., Yu Q; Robert Frederick Smith School of Chemical and Biomolecular Engineering, Cornell University, Ithaca, New York 14853, United States. |
| المصدر: | ACS sensors [ACS Sens] 2024 Jul 26; Vol. 9 (7), pp. 3594-3603. Date of Electronic Publication: 2024 Jun 24. |
| نوع المنشور: | Journal Article |
| اللغة: | English |
| بيانات الدورية: | Publisher: American Chemical Society Country of Publication: United States NLM ID: 101669031 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 2379-3694 (Electronic) Linking ISSN: 23793694 NLM ISO Abbreviation: ACS Sens Subsets: MEDLINE |
| أسماء مطبوعة: | Original Publication: Washington, DC : American Chemical Society, [2016]- |
| مواضيع طبية MeSH: | Surface Plasmon Resonance*/methods , Extracellular Vesicles*/chemistry, Animals ; Immunoassay/methods ; Mice ; Tetraspanin 28/analysis ; Tetraspanin 28/chemistry ; Tetraspanin 28/metabolism ; Antibodies, Immobilized/immunology ; Antibodies, Immobilized/chemistry ; Biosensing Techniques/methods ; Membrane Proteins/chemistry |
| مستخلص: | Extracellular vesicles (EVs) are preeminent carriers of biomarkers and have become the subject of intense biomedical research for medical diagnostics using biosensors. To create effective EV-based immunoassays, it is imperative to develop surface chemistry approaches with optimal EV detection targeting transmembrane protein biomarkers that are not affected by cell-to-cell variability. Here, we developed a series of immunoassays for the detection of EVs derived from mouse monocyte cells using surface plasmon resonance (SPR) biosensors. We chemically immobilized antibodies onto mixed self-assembled monolayers of oligo ethylene glycol (OEG) alkanethiolates with carboxylic and hydroxylic terminal groups. The effects of antibody clonality (monoclonal vs polyclonal) and antibody surface coverage in targeting EVs via CD81 tetraspanins were investigated. We determined binding kinetic parameters, establishing trends from steric hindrance effects and epitope recognition properties of antibodies. Our results indicate that a 40% surface coverage of polyclonal antibodies covalently linked onto a mixed SAM with 10% of terminated -COOH groups yields a promising approach for EV detection with a linear range of 1.9 × 10 8 -1.9 × 10 9 EVs/mL and a limit of detection of 5.9 × 10 6 EVs/mL. This optimal immunoassay exhibits a 1.92 nM equilibrium dissociation constant for bound EVs, suggesting a high binding affinity when CD81 is targeted. Our study provides important insights into surface chemistry development for EV detection targeted via transmembrane protein biomarkers using antibodies, which has promising applications for disease diagnostics. |
| فهرسة مساهمة: | Keywords: biomarkers; biosensors; extracellular vesicles; self-assembled monolayer; surface plasmon resonance; tetraspanin |
| المشرفين على المادة: | 0 (Tetraspanin 28) 0 (Antibodies, Immobilized) 0 (Membrane Proteins) |
| تواريخ الأحداث: | Date Created: 20240624 Date Completed: 20240726 Latest Revision: 20240726 |
| رمز التحديث: | 20240726 |
| DOI: | 10.1021/acssensors.4c00558 |
| PMID: | 38912608 |
| قاعدة البيانات: | MEDLINE |
Find this issue from the American Chemical Society | تدمد: | 2379-3694 |
|---|---|
| DOI: | 10.1021/acssensors.4c00558 |