Biofunctionalized Nanoconjugates: Design, Production, and Characterization

Integrated service to create and optimize antibody-functionalized nanoconjugates for reliable diagnostics and nanomedicine.

Scientific Leader / leaders of the project: Prof. M. Pilar Marco (U2, CAbS) and Dr. Carlos Rodríguez (U12)
Coordinator of the project: Nuria Pascual (U2) and Susana Vilchez (U12)

Industrial gap/problem covered

Many diagnostic and nanomedicine projects fail to progress beyond the prototype stage because the recognition element and the nanostructured carrier are developed separately. In practice, antibody generation, hapten design, and immunoassay development require strong expertise in antigen selection and bioconjugation, while colloidal systems intended for diagnostics, drug delivery, or theranostics must be optimized for size, surface charge, stability, formulation robustness, and reproducibility. This gap becomes even more critical when the final product is an antibody-functionalized nanoprobe, a liposomal or micellar assay reagent, or a bioconjugate intended for point-of-care use. 

Description

This combined service offers an integrated workflow that connects the production of custom antibodies and immunoreagents (U2) with the physicochemical characterization of nanostructured liquids, dispersions, and nanoassemblies (U12). U2 contributes the design of haptens and peptides, antibody generation and purification, immunoassay development, and bioconjugation expertise. U12 adds advanced characterization of colloidal systems, including DLS/SLS, zeta potential, colloidal stability, phase behavior, surface/interfacial tension, viscosity, microscopy, and SEM. Together, both units provide a seamless route to develop, optimize, and validate antibody-based nanoprobes, nanodiagnostics, and nanoformulations with higher robustness and translational potential.

Examples and scientific publications implementing this Cutting-Edge Biomedical Solution:

1. Nanoplasmonic biosensor for acenocoumarol monitoring. NANBIOSIS reports a collaboration in which U2 produced highly specific polyclonal antibodies and the antibody-based biosensor was integrated into a gold nanostructure platform for plasma monitoring of acenocoumarol.
   • Related Publication: Peláez, E. C., Estevez, M.-C., Portela, A., Salvador, J.-P., Marco, M.-P., & Lechuga, L. M. (2018). Nanoplasmonic biosensor device for the monitoring of acenocoumarol therapeutic drug in plasma. Biosensors and Bioelectronics, 119, 149–155.
2. Characterization of nanomedicines / PLGA nanoparticles. The U12 infrastructure page describes the characterization of nanostructured systems and includes an example where DLS and microscopy were used to study MNP-loaded PLGA nanoparticles for theragnostic applications (Monge et al. 2020).
   • Related publication: Marta Monge Azemar, Cristina Fornaguera, Carmen Quero, Aurora Dols-Perez, Gabriela Calderó, Santiago Grijalvo, María-José García-Celma, Carlos Rodriguez-Abreu, Conxita Solans, “Functionalized PLGA nanoparticles prepared by nano-emulsion templating interact selectively with proteins involved in the transport through the blood-brain barrier” (2020). European Journal of Pharmaceutics and Biopharmaceutics.

Services involved:

The steps and services involved in this cutting-edge solution are the following:

1. Target definition, epitope/hapten selection and project design (U2): Selection of the biomarker, peptide epitope or hapten, and definition of the most appropriate antibody format and intended nanostructure.
2. Synthesis of haptens, immunogens and conjugates (U2): Preparation of peptide or small-molecule haptens, carrier conjugation, and pre-immunization quality control
3. Production and purification of monoclonal/polyclonal antibodies (U2): Hybridoma generation, antibody screening, purification, and selection of the best binders for the intended application
4. Functionalization of nanostructures with antibodies or immunoreagents (U2 + U12): Coupling of antibodies to nanoparticles, liposomes, micelles, emulsions or other colloidal systems, with iterative optimization of the conjugation conditions
5. Physicochemical characterization of the nanoformulation (U12-S08): Size distribution, zeta potential, CMC, phase behavior, solubilization, turbidity, density, viscosity, surface/interfacial tension, hydrophobicity, microscopy and SEM
6. Stability and performance optimization (U2+ U12): Assessment of colloidal stability, compatibility with assay conditions, and adjustment of formulation parameters until a robust and reproducible product is obtained

• U2
• U12

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CAbS (U2) is managed by the scientific team of the Nb4D group (Nb4D) with high expertise in hapten synthesis, conjugation and antibody production.We have an extensive track record of successful projects in developing polyclonal and monoclonal antibodies against small molecules and peptidesfor the development of diagnostic tools. These include, for example, the evaluation of carcinogenic processes, cardiovascular diseases, and infectious diseases.

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Our Unit 12 is located at the Institute of Advanced Chemistry of Catalonia (IQAC-CSIC), in Barcelona and it is coordinated by the Colloidal and Interfacial Chemistry Group, led by Dr. Carlos Rodríguez. The unit is focused on the characterization of nanostructures in liquids and liquid/liquid interfaces, such as micelles, liposomes, micro- and nano-emulsions, and nanoparticles. The unit features state-of-the-art instrumentation for the determination of various parameters such as size distribution and zeta potential (surface charge), critical micelle concentration (CMC), solubilization capacity, osmolarity, surface and interfacial tension, wettability, refractive index, turbidity, colloidal stability, density, viscosity and other rheological parameters (shear and elastic moduli, critical yield stress, etc.) The Unit 12 has implemented and maintains the ISO9001 certification forstandard quality control system.​