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Biomolecules production - Services

Biomolecules production – Services

U29-S02. Modification of oligonucleotides. (Remote) OUTSTANDING

Modification of oligonucleotides

This service is dedicated to the custom synthesis of oligonucleotide conjugates including oligonucleotides carrying lipids, amino acids, peptides or carbohydrates. In addition, they may include other modifications such as phosphorothioate linkages, 2’-O-methyl-RNA, 2’-O-MOE-RNA, 2′-F-RNA, Locked nucleic acids (LNA), modified nucleotides. The oligonucleotides will be prepared in 1 micromol scale. In most cases, the preparation of the conjugates will require the preparation of oligonucleotides carrying reactive groups such as; amino or thiol groups, which will be subjected to a post-synthetic modification. Purification and characterization will include reversed-phase HPLC analysis and mass spectrometry (MALDI-TOF).

Modification of oligonucleotides during and post synthesis to meet user requirements:

  • Conjugation of oligonucleotides with fluorophores (fluoresceine, Cy3, Ct5, etc..) and other types of small molecules such as biotine, lipids.
  • Conjugation to peptides.
  • Phosphorothioate linkages
  • Modified backbones such as locked nucleic acids (LNA), 2’-O-alkyl-RNA, etc..
  • Modified nucleobases: 2-aminopurine, 5-methyl-dC, etc…
  • 5’, 3’-modifications such as 5’-, 3’-amino, 5’-, 3’-thiol, etc..

Customer benefits

The service benefits from the 40-years’ experience of the researchers participating in the service, with hundreds of scientific communications on improving the methodology used for the synthesis of modified oligonucleotides. This includes fatty acid derivatives, amino acids, cell penetrating peptides and carbohydrates not available from other services. In addition, the service can develop customized solutions for molecules not addressed in the bibliography.

Target customer

The primary audience are research groups and companies working on gene therapy and gene inhibition in the early stages of preclinical development.

References

1) “Aptamer-peptide conjugates as a new strategy to modulate human α-thrombin binding affinity”. Aviñó, A. et al. Biochim. Biophys. Acta (General subjects), 1863, 1610-1630 (2019).
2) “Synthesis of oligonucleotides carrying amino-lipid groups at the 3’-end for RNA interference studies”. Grijalvo, S. et al. J. Org. Chem., 75, 6806-6813 (2010).
3) “Synthesis and evaluation of 3’ oleyl-oligonucleotide conjugates as potential cellular uptake enhancers”. Navarro, N. et al. SYNLETT, in press (2024). doi: 10.1055/s-0042-1751528.

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U29-S03. Special nucleotides for oligonucleotide synthesis. (Remote) OUTSTANDING

Special nucleotides for oligonucleotide synthesis

This service is dedicated to the custom synthesis of modified nucleotides such as nucleoside monophosphates or triphosphates as well as special phosphoramidites or functionalized solid supports functionalized with small molecules resistant to ammonia deprotection for the preparation of oligonucleotide conjugates.

Special nucleotides for oligonucleotide synthesis

For those services identified as outstanding, at least 20% of their capacity is open under competitive access. See Annex 1 of ACCESS PROTOCOL (provided by Nanbiosis) for details on % of openness for each service

Customer benefits

The service benefits from the 40-years’ experience of the researchers participating in the service with hundreds of scientific communications on improving the methodology used for the synthesis of modified oligonucleotides. This includes a range of modified nucleotides and terminal modifications not available in other services. In addition, the service can develop customized solutions for molecules not covered in the bibliography.

Target customer

The primary audience are research groups and companies involved in gene therapy, gene silencing and the development of nucleic acid-based diagnostic tools.

References

1) “Oligonucleotides containing 1’-aminomethyl or 1’-mercaptomethyl-2’-deoxy-D-ribofuranoses: Synthesis, purification, characterization and conjugation with fluorophores and lipids”. Martín-Nieves, V. et al. Bioconjugate Chem, 32, 350-366 (2021).
2) “Efficient bioactive oligonucleotide-protein conjugation for cell-targeted cancer therapy”. Aviñó, A. et al. Chemistry Open, 8, 382-387 (2019).
3) “Thioctic acid derivatives as building blocks to incorporate DNA oligonucleotides onto gold nanoparticles”. Pérez-Rentero, S. et al. Molecules, 19, 10495-10523 (2014).

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U29-S01. Synthesis of oligonucleotides and characterization (On-site & Remote). OUTSTANDING

Synthesis of oligonucleotides and characterization

This service is dedicated to the custom synthesis of modified oligonucleotides including phosphorothioate linkages, 2’-O-methyl-RNA, 2’-O-MOE-RNA, 2′-F-RNA, Locked nucleic acids (LNA), modified nucleotides and several others. The oligonucleotides will be prepared on a 1 micromol scale. Purification and characterization will include reversed-phase HPLC analysis and mass spectrometry (MALDI-TOF).

Synthesis of oligonucleotides at various different scales (100 microg to 5 mg) and purification using HPLC and/or desalting.

Customer benefits

The service benefits from the 40-years’ experience of the researchers involved in the service, with hundreds of scientific communications on improving the methodology used for the synthesis of modified oligonucleotides. This expertise includes a range of modified nucleotides and terminal modifications that are not available in other services. In addition, the service can develop customized solutions for molecules not covered in the bibliography.

Target customer

The primary audience are research groups and companies involved in mutagenesis, gene therapy,  gene inhibition, the development of nucleic acid-based diagnostic tools, or the study of nucleic acid structure and nucleic acid-protein interaction.

References

1) “Detection of SARS-CoV-2 virus by Triplex Enhanced Nucleic Acid Detection Assay (TENADA)”, Aviñó, A., et al. Int. J. Mol. Sci., 23, 15258 (2022).
2) “Properties of parallel tetramolecular G-quadruplex carrying N-acetylgalactosamine as potential enhancers for oligonucleotide delivery to hepatocytes”. Clua, A. et al. Molecules, 27, 3944, (2022).
3) “Chemical modifications in nucleic acids for therapeutic and diagnostic applications”. Fàbrega, C., Aviñó, A., Eritja, R. The Chemical Record, 22, e202100270 (2022).

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U28-S07. Transmission Electron Microscopy (TEM)

Transmission Electron Microscopy (TEM)

The Electron Microscopy Service offers access to transmission electron microscopy (TEM), cryo-TEM and electron tomography, high resolution scanning electron microscopy (SEM), and environmental SEM. A special emphasis has been made on high-end sample preparation techniques through cryo-immobilization. All the equipment has been configured to provide its best for biological and nanomedicine applications.

Customer benefits

We have SOPs and ISO9001 certification. We also have specialist technicians for the use of the equipment.

This service is essential to:

  • Cellular characterization: cell structures and ultrastructures visualization and its relationship to function are the most important contributions of EM to cell biology.
  • Tissue characterization: electron microscopy has a role in the characterization of interactions between cells and with other components inside tissues.
  • Biomaterials characterization: electron microscopy plays a double role in biomaterial characterization On one hand, it provides structural and compositional information on the materials engineered to be used inside biological systems, favouring its development On the other hand, it allows us to visualize their interactions Functionalized polymeric and metallic nanoparticles for drug delivery, dental implants, bone plates and cements, and artificial tissues are among the biomaterials that can benefit from the information provided by EM.
  • Macromolecular complexes characterization: structural biology can benefit from electron microscopy to determine 3D structures of macromolecular complexes.
  • Negative staining is also a very valuable tool for 2D-3D characterization.

Target customer

Any company or research group interested in:

  • Transmission electron microscopy of resin-embedded and non-embedded samples.
  • Cryo-fixation of samples at high pressure (High-Pressure Freezing).
  • Resin embedding of samples at room temperature (conventional) or low temperature (freeze-substitution) for ultramicrotomy.
  • Ultramicrotomy of resin-embedded sections (semithin and ultrathin sections).
  • Data and image processing.
  • Technical advice for users regarding the selection of protocols and procedures for experiments involving electron microscopy.

References

  1. Gómez-González E, González-Mancebo D, Núñez NO, Caro C, García-Martín ML, Becerro AI, Ocaña M. Lanthanide vanadate-based trimodal probes for near-infrared luminescent bioimaging, high-field magnetic resonance imaging, and X-ray computed tomography. J Colloid Interface Sci. 2023 Sep 15;646:721-731. doi: 10.1016/j.jcis.2023.05.078. Epub 2023 May 18. PMID: 37229990.
  2. Caro C, Guzzi C, Moral-Sánchez I, Urbano-Gámez JD, Beltrán AM, García-Martín ML. Smart Design of ZnFe and ZnFe@Fe Nanoparticles for MRI-Tracked Magnetic Hyperthermia Therapy: Challenging Classical Theories of Nanoparticles Growth and Nanomagnetism. Adv Healthc Mater. 2024 Feb 2:e2304044. doi: 10.1002/adhm.202304044. Epub ahead of print. PMID: 38303644.

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U3-S04. Peptide libraries (Remote) OUTSTANDING

Peptide libraries.

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U3-S03. Special amino acids for peptidomimetics synthesis (Remote) OUTSTANDING

Special amino acids for peptidomimetics synthesis (Remote) OUTSTANDING

– Synthesis of N-alkyl amino acids and other special amino acids
– Synthesis of peptoids (N-alkylglycine oligomers).
– Synthesis of β-peptide, γ- peptides, β,γ-peptides and α,β-peptides
– Synthesis of peptidomimetics
– Synthesis of hybrid-heterocycle- peptides

Customer benefits

  • Extensive experience in the synthesis of peptide oligomers from non-natural amino acids such as β-peptides, γ-peptides, β,γ-peptides and α,β-peptides.
  • Experience with peptides and peptide mimetics in solid phase and in solution.
  • Development of various strategies for large-scale production (hundreds of mg) of peptidomimetics.

Target customer

  • Research groups (drug delivery, molecular biology, pharmacology, nanotechnology, biotechnology)
  • Companies (biotech and pharma companies).

References

  • Hybrid cyclobutane/proline-containing peptidomimetics: the conformational constraint influences their cell-penetration ability. Illa, Ona ; Ospina, Jimena; Sanchez-Aparicio, Jose-Emilio; Pulido, Ximena ; Abengozar, Maria Angeles; Gaztelumendi, Nerea; Carbajo, Daniel; Nogues, Carme; Rivas, Luis ; Marechal, Jean-Didier; Royo, Miriam ; Ortuño, Rosa M. International Journal of Molecular Sciences (2021), 22, 5092.
  • Chiral cyclobutane-containing cell-penetrating peptides as selective vectors for anti-Leishmania drug delivery systems. Illa, Ona ; Olivares, Jose-Antonio; Gaztelumendi, Nerea; Martinez-Castro, Laura; Ospina, Jimena; Abengozar, Maria-Angeles; Sciortino, Giuseppe ; Marechal, Jean-Didier; Nogues, Carme; Royo, Miriam; Rivas, Luis; Ortuno, Rosa M. International Journal of Molecular Sciences (2020), 21, 7502.
  • A solid-phase combinatorial approach for indoloquinolizidine-peptides with high affinity at D1 and D2 dopamine receptors. Molero, Anabel; Vendrell, Marc; Bonaventura, Jordi; Zachmann, Julian; Lopez, Laura; Pardo, Leonardo; Lluis, Carme; Cortes, Antoni; Albericio, Fernando; Casado, Vicent; Royo, Miriam. European Journal of Medicinal Chemistry (2015), 97, 173-180.
  • Efficient γ-amino-proline-derived cell penetrating peptide-superparamagnetic iron oxide nanoparticle conjugates via aniline-catalyzed oxime chemistry as bimodal imaging nanoagents. Cavalli, Silvia; Carbajo, Daniel; Acosta, Milena; Lope-Piedrafita, Silvia; Candiota, Ana Paula; Arus, Carles; Royo, Miriam; Albericio, Fernando. Chemical Communications (2012), 48, 5322-5324.

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U2-S08. Inmunochemical methods development

Inmunochemical methods development

The service offers custom tailored immunoassay development. It involves designing and optimizing assays for detecting specific analytes (such as proteins, antibodies, or other biomolecules) in biological samples. The service includes identifying necessary reagents, establishing assay parameters, and validating the assay’s performance.

Customer benefits

  • Tailored Solutions: By utilizing our custom tailored immunoassay development service, customers gain access to highly specialized expertise and resources, resulting in the creation of assays optimized for their unique requirements.
  • Enhanced Sensitivity: The service ensures optimal sensitivity, allowing for the detection of analytes at low concentrations.
  • Time and Cost Savings: Outsourcing immunoassay development to experts saves time and resources compared to in-house development.
  • Quality Assurance: Rigorous validation ensures reliable results, enhancing confidence in research outcomes.

Target customer

  • Research Institutions: Universities, research centers, and laboratories engaged in biomedical research.
  • Biotech Companies: Organizations involved in drug discovery, diagnostics, and therapeutic development.
  • Pharmaceutical Companies: Those working on antibody-based therapies or vaccines.
  • Healthcare Providers: Hospitals and clinics conducting specialized research.
  • Food & Environmental Quality Control Companies: Entities involved in:
    • Food Contaminant Detection: Developing antibody-based
      tests for identifying foodborne pathogens and toxins.
    • Environmental Contaminant Detection: Creating antibody-based assays for detecting environmental pollutants in air, water, and soil.

Additional information

Selected references:

  • G. Colom, A. Hernández-Albors, J. Barallat, A. Galan, A. Bayes-Genis, J.-P. Salvador, M.-P. Marco. A multiplexed immunochemical microarray for the determination of cardiovascular disease biomarkers. Microchim. Acta, 191, 54, 2023.
  • C. Adrover-Jaume, A. Clemente, B. Rodríguez-Urretavizcaya, L. Vilaplana, M.-Pilar Marco, E. Rojo-Molinero, A. Oliver, R. de la Rica. A paper biosensor for overcoming matrix effects interfering with the detection of sputum pyocyanin with competitive immunoassays. Microchim acta, 190, 441, 2023.
  • E. Montagut, J. Raya, M.-T. Martín Gómez, L. Vilaplana, B. Rodríguez- Urretavizcaya, M.-P. Marco. An Immunochemical Approach to detect the Quorum Sensing-Regulated Virulence Factor 2-Heptyl-4- Quinoline N-Oxide (HQNO) produced by Pseudomonas aeruginosa Clinical Isolates. Microbiol. Spect., 10(4), 1-12, 2022.
  • E. Montagut, G. Acosta, F. Albericio, M. Royo, G. Godoy-Tena, A. Lacoma, C. Prat, J.-P. Salvador, M.-P. Marco. Direct Quantitative Immunochemical Analysis of the Autoinducer Peptide IV (AIP-IV) for Diagnosing and Stratifying Staphylococcus aureus infections. ACS Infect. Dis., 8(3), 645-656, 2022.
  • B. Rodriguez-Urretavizcaya, N. Pascual, C. Pastells, M. T. Martin-Gomez, Ll. Vilaplana, M.-P. Marco. Diagnostic and Stratification of Pseudomonas aeruginosa Infected Patients by Immunochemical Quantitative Determination of Pyocyanin from Clinical Bacterial Isolates. Frontiers in Cell. Infect. Microbiol., 11, 786929, 2021. DOI: 10.3389/fcimb.2021.786929
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U2-S07. Antibody purification (Remote) OUTSTANDING

The purification of the antibodies includes the isolation of the antibody from the serum (polyclonal antibodies) or culture supernatant from hybridoma cell lines (monoclonal antibodies). The services offers previous advise on the different possibilities for purification according to the needs of the user.

The methods of purification that we offer vary from crude methods (precipitation of the proteins from the sample including any antibody present) to general purification (purification by affinity of certain classes of antibodies without taking account of the specificity to the antigen) to specific (purification by affinity of the antibodies which join specifically to a definite antigen).

  • Ammonium sulfate precipitation
  • Protein A, protein G affinity purification
  • Specific antigen affinity purification
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U2-S06. Hybridoma cell culture antibody production (Remote) OUTSTANDING

Hybridoma cell culture in ultralow serum medium. We use CELLine™  Bioreactor Technologys to produce monoclonal antibodies.

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U2-S05. Monoclonal Antibody development (Remote) OUTSTANDING

The mAb development contains the following phases:

  1. Immunisation of mice, titre determination in the antiserum.
  2. Cultivation of the myeloma cell line, cell fusion, selection in HAT medium and screening.
  3. Cloning , screening and cultivation of positive cultures. Further cloning and obtaining stable clones.
  4. Expansion and Cryo conservation of the selected clones. Delivery of cell culture supernatants and frozen hybridoma cells.
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