U2. Custom Antibody Service (CAbS)

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

U2-S07. Antibody purification

Antibody purification

Antibody purification involves the selective enrichment or specific isolation of antibodies from various sources, such as serum (for polyclonal antibodies), or cell culture supernatant (for monoclonal antibodies).

The main methods used for antibody purification are:

• Physicochemical Fractionation: This method includes differential precipitation, size-exclusion, or ion exchange chromatography, based on size, charge, or other shared chemical characteristics It separates proteins that includes the immunoglobulins (antibodies) based on size, charge, or other shared chemical characteristics
• Class-Specific Affinity: In this approach, specific antibody classes (e.g., IgG) are bound to solid phases using immobilized biological ligands (such as proteins A or G). This method purifies all antibodies of the target class without considering antigen specificity.
• Antigen-Specific Affinity: Here, only antibodies in a sample that bind to a particular antigen are purified. Affinity chromatography is performed using a column packed with immobilized antigen (the same antigen used for immunization). Only antibodies that bind to the antigen are isolated. This method provides a higher yield of antigen-specific antibodies than Protein A/G affinity chromatography, though the total amount of recovered antibodies is lower.

Customer benefits

• High Purity: Antibody purification ensures that the isolated antibodies are of high purity, minimizing contamination by other proteins.
• Specificity: By using antigen-specific affinity purification, researchers obtain antibodies that specifically recognize their target antigen.
• Consistent Results: Purified antibodies lead to more consistent and reliable experimental results.
• Diagnostic and Therapeutic Applications: Purified antibodies are essential for diagnostic assays, therapeutic development, and research applications.
• Quality: We operate under ISO 9001 quality standards, ensuring consistent, reliable antibody production.

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:

• Giovanna Roncador; Pablo Engel; Lorena Maestre; et al; Alison H.Banham., Nuria Pascual 2016. The European antibody network’s practical guide to finding and validating suitable antibodies for research. mAbs. Taylor & Francis Online. 8-1, pp.27-36.
• 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

U2-S06. In vitro antibody production

In vitro antibody production

We perform the production of monoclonal antibodies in vitro.
Hybridoma cells are first expanded in culture flask or high density culture devices (for larger amounts of antibody) to get the optimal cell density needed for the production.
The classical medium of production includes fetal calf serum (ultralow IgG, FCS) but our expertise allows us to provide a completely customized service according to your needs, without using FCS.
Supernatant containing the monoclonal antibodies is then periodically collected until the expected quantity of antibodies is obtained.

Depending on the nature of the sample, the expected antibody concentrations are for :­

• Culture in flask: from 5 to 20 µg antibody/mL supernatant
• ­ Culture in high density device : up to 1 mg antibody/mL supernatant

Customer benefits

• Customization: The service can be tailored to meet the specific needs of each customer, allowing for personalized antibody production without using fetal calf serum (FCS).
• Scalability: The service can accommodate both flask cultures and high-density devices, providing flexibility for different antibody quantities.
• ISO 9001 Certification: We operate under ISO 9001:2015 quality standards, ensuring consistent and reliable antibody production.

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:

• Giovanna Roncador; Pablo Engel; Lorena Maestre; et al; Alison H.Banham., Nuria Pascual 2016. The European antibody network’s practical guide to finding and validating suitable antibodies for research. mAbs. Taylor & Francis Online. 8-1, pp.27-36
• 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

U2-S05. Monoclonal Antibody development

Monoclonal Antibody development

To develop customized monoclonal antibodies, we adapt our protocols at each critical stage of antibody production. This includes designing the antigen, immunization protocol, and screening design of hybridoma cell supernatants. The initial step involves immunizing mice or rats with a specific antigen. We can develop monoclonal antibodies against various antigen types, such as proteins, peptides, small molecules, or cells. The immunization process typically spans 10-12 weeks. The animal with the most robust immune response undergoes a final immunization to stimulate antibody-producing cells. Spleen cells are isolated and fused with immortal myeloma cell lines to create hybridoma cells. The chosen hybridoma clones are then subcloned to ensure cell stability and maintain their monoclonal characteristics. Finally, the isolated cells are expanded and cryopreserved.

Customer benefits

• Versatility: Monoclonal antibodies can be used in diagnostics, therapeutics, and research.
• Customization: Protocols can be adapted to meet specific needs
• Quality Assurance: Our processes adhere to the ISO 9001 quality requirements.

Target customer

• Organizations involved in research and development, pharmaceutical companies, academic institutions, and diagnostic laboratories benefit from monoclonal antibody development.
• Researchers studying diseases, drug targets, and immune responses rely on mAbs for their work.

Additional information

Selected references:

• Giovanna Roncador; Pablo Engel; Lorena Maestre; et al; Alison H.Banham., Nuria Pascual 2016. The European antibody network’s practical guide to finding and validating suitable antibodies for research. mAbs. Taylor & Francis Online. 8-1, pp.27-36
• 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

U2-S04. Polyclonal Antibody Production

Polyclonal Antibody Production

Firstly, your project is discussed with our team, we help you to make the right decisions and personalize the process according to your goals. For the production of polyclonal antibodies, we select New Zealand white rabbits. Additionally, we offer the development of polyclonal antibodies in mice and rats.
We can generate policlonal antibodies towards a variety of antigens including small molecules, peptides, proteins, and cells.
Our standard immunization protocol includes monthly inoculations with Freund’s adjuvant over a 6-month period. To assess immune response progress, blood samples are taken 10 days after the second inoculation to obtain serum and determine antibody titers using ELISA. Preimmune serum is collected before the first injection for controls. At the end of the process, complete blood collection from the animal is performed surgically under anesthesia, resulting in 50-70 mL of hyperimmune serum per animal. The service provides 5 mL of bleed serum from each test for screening in customer labs.
We can follow your personalized immunization program in addition to our well-established ones.

Customer benefits

• Scientific Consultation: We provide thorough consultation before the project begins.
• Customized Project Proposals: We tailor project proposals to meet your specific requirements.
• Flexible Workflow: Adjustments can be made to the workflow based on results and your specific requests.

Target customer

Organizations involved in research and development, particularly those seeking reliable and customized polyclonal antibody development services, will find our offering essential for their scientific endeavors.

Additional information

Selected references:

• 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.
• ­G. Colom, J.-P. Salvador, G. Acosta, M. Royo, M.-P. Marco. Competitive ELISA for N-Terminal pro-Brain Natriuretic Peptide (NT-proBNP) determination in human plasma. Analyst, 145, 6719-6727, 2020.

U2-S03. Preparation of Bioconjugates and Molecular Probes

Preparation of Bioconjugates and Molecular Probes

The preparation of bioconjugates and molecular probes involves a diverse array of techniques and strategies aimed at functionalizing biomolecules for various applications. This process encompasses the labeling of antibodies, haptenized proteins, and enzymes, as well as the biotinylation and fluorescent labeling of probes, and the conjugation of biomolecules with nanoparticles and other entities.

Customer benefits

• To develop antibodies against small molecules (preparation of immunization bioconjugate)
• Enhanced Functionality: Bioconjugation allows for specific binding of biomolecules, such as antibodies or peptides, to other entities like molecular probes or nanoparticles. This expands the applications and functionalities of biomolecules.
• Improved Sensitivity and Specificity: Bioconjugates can enhance the sensitivity and specificity of detection techniques, such as immunohistochemistry or flow cytometry.
• Diverse Applications: Bioconjugates find application in research, diagnostics, and therapy, benefiting research laboratories, hospitals, and pharmaceutical companies.

Target customer

• Researchers and Scientists: Satisfy the need to label biomolecules for research studies.
• Pharmaceutical and Biotechnology Companies: For creating therapeutic bioconjugates, imaging markers or bioconjugates for immunizations of non-immunogenic small molecules, between other.

Additional information

Selected references:

• 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.
• 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.
• J. Marrugo-Ramírez, M. Rodríguez-Núñez, M.-P Marco, M. Mir, J. Samitier. Kynurenic Acid Electrochemical Immunosensor: Blood-Based Diagnosis of Alzheimer’s Disease. Biosensors, 11(1), 20, 2021.
• E. J. Montagut, Ll. Vilaplana, M.T. Martin-Gómez, M.-P. Marco. A High Throughput Immunochemical Method to Assess 2-Heptyl-4-Quinolone Quorum Sensing Molecule as Potential Biomarker. ACS Infect. Dis., 6(12), 3237-3246, 2020.
• M. Broto, R. McCabe, R. Galve, M.-P. Marco. A high-specificity immunoassay for the therapeutic drug monitoring of ciclophosphamide. Analyst, 144, 5172-5178, 2019.

U2-S02. Hapten design and synthesis

Hapten design and synthesis

Haptens are small molecules capable of eliciting an immune response when conjugated to a carrier protein. They serve as antigens for antibody production. The service designs and produces tailored haptens to match the target analyte, ensuring the desired features of the resulting antibody.

This process includes:

• Selecting the most suitable position on a drug molecule to attach a linker, maximizing specificity, sensitivity, and immunogenicity of the immunogen
• Performing total synthesis to create the hapten with a linker when the parent drug molecule lacks a functional group suitable for conjugation synthesis.
• Designing and synthesizing the linker with the optimal length.

Customer benefits

Some specific advantages that customers gain by utilizing this service include:

• Adaptation of haptens to meet specific customer needs.
• Production of highly specific and  antibodies.
• Added value in research and development by obtaining personalized immunological tools.
• Essential for organizations involved in drug discovery, diagnostics, and therapy.

Target customer

Target customer Researchers, scientists, and professionals involved in research and development in various fields including food safety and environmental control, biomedicine, diagnostics, drug discovery, and therapeutic development.

Additional information

Selected references:

• 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.
• 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.
• J. Marrugo-Ramírez, M. Rodríguez-Núñez, M.-P Marco, M. Mir, J. Samitier. Kynurenic Acid Electrochemical Immunosensor: Blood-Based Diagnosis of Alzheimer’s Disease. Biosensors, 11(1), 20, 2021.
• E. J. Montagut, Ll. Vilaplana, M.T. Martin-Gómez, M.-P. Marco. A High Throughput Immunochemical Method to Assess 2-Heptyl-4-Quinolone Quorum Sensing Molecule as Potential Biomarker. ACS Infect. Dis., 6(12), 3237-3246, 2020.
• M. Broto, R. McCabe, R. Galve, M.-P. Marco. A high-specificity immunoassay for the therapeutic drug monitoring of ciclophosphamide. Analyst, 144, 5172-5178, 2019.