Are you struggling to find reliable antibodies against G-protein-coupled receptors (GPCRs) for your scientific or clinical projects? At ProteoGenix, we understand the challenges of developing high-quality GPCR antibodies. That’s why we use advanced techniques and rigorous quality control testing to ensure the reproducibility and specificity of every GPCR antibody we customize. During the whole custom antibody development process, from antigen design and production to antibody generation and purification, our team of experts works closely with you through predefined project milestones to ensure that your antibody is built on time and on budget. Contact us today to see how our custom monoclonal antibodies can help you achieve your research goals.

Find the Perfect Method for Making your custom GPCR Antibody

ProteoGenix uses one of three different antibody engineering strategies to make custom monoclonal antibodies that bind GPCR antibodies. These include creating hybridoma cell lines, isolating anti-GPCR-specific B-cells, and screening for specific anti-GPCR antibodies using phage display. ProteoGenix can assist you in determining which method is best suited to your project and budget.

Antibody Phage Display

Antibody phage display is a method that involves obtaining peripheral blood mononuclear cells (PBMCs) from either immunized animal hosts (immune libraries) or unimmunized hosts (naïve libraries). After PBMC collection, the antibody gene segments are converted into cDNA and cloned into a phagemid to create an antibody-phage fusion protein, which displays the antibody on the outer surface of the bacteriophage.
Next, the target antigen is cloned and immobilized to a surface so the antibody on the phage surface can bind the immobilized antigen, a process called biopanning. Once the antibody binds to the antigen, the DNA inside the bacteriophage, containing the antibody-related genes, is extracted, sequenced, and expressed to confirm its efficacy. Learn more about our antibody phage display service.

Naive Library Workflow

Antigen procurement or design and production

  • Our experts immunize animal hosts using GPCRs in micelles, nanodiscs, or whole cells that express your GPCR of interest.

Library screening and biopanning

  • Screening of naive or immune library for antigen binders

ELISA screening of single phage binders

  • Further validate binders by ELISA screening until at least 3-10 different binders have been identified.

DNA extraction & antibody sequencing

Immune Library Workflow

Antigen procurement or design and production

  • Our experts immunize animal hosts using GPCRs in micelles, nanodiscs, or whole cells that express your GPCR of interest.

Immune library construction

  • PBMC isolation
  • RNA extraction and cDNA synthesis
  • VH and VL PCR amplification
  • Library construction and quality control

Library screening and biopanning

  • Screening of naive or immune library for antigen binders

ELISA screening of single phage binders

  • Further validate binders by ELISA screening until at least 3-10 different binders have been identified.

DNA extraction & antibody sequencing

Why Antibody Phage Display is Ideal for Making Human Therapeutic Antibodies

Antibody phage display is a valuable technique for producing monoclonal antibodies that target cytotoxic antigens, rare antigens (or epitopes), or antigens with low immunogenicity. In addition, antibody phage display is particularly useful for making custom antibodies used clinically because of ProteoGenix’s flexible cancer and autoimmune human phage display libraries. These new libraries offer our clients the flexibility to quickly, identify unique human antibodies that bind their GPCR of interest cost-effectively.
If your biomedical research lab or pharmaceutical company aims to expedite human immunotherapy projects aimed at treating cancer, autoimmunity, or other diseases, then antibody phage display technology is the ideal choice.

Phage Display Has No Antibody Species Limitations

Antibody phage display is a powerful technique for custom antibody generation because there is no species limitation. Whether you’re interested in making unique anti-GPCR antibodies to perform in a specific biomedical research experiment or you need to build complex immunotherapy, ProteoGenix can meet your needs. Our team can immunize rabbits, mice, and even camelids to generate custom anti-GPCR antibodies that target your favorite GPCR with precision. We can also screen your GPCR of interest using our pre-built naive human autoimmune and cancer libraries to quickly identify antibodies that bind rare GPCR epitopes. Our comprehensive approach ensures that you receive high-quality, fully customized antibodies for your specific research needs.

Phage Display Delivers Antibody DNA Sequences, That You Own

Once we discover at least three different antibodies that bind your GPCR with precision, we will deliver the antibody DNA (cDNA) sequences to you. These sequences become your intellectual property (IP) giving your institution or company the following benefits:

  • The right to patent the antibody.
  • Access to potential revenue streams from licensing agreements, royalties, or other financial arrangements.
  • Greater control over the antibody’s development, production, and distribution.
  • Protecting the antibody from being copied or exploited by competitors and provide legal recourse in case of infringement.
  • Enhancing your scientific reputation and credibility as a leader in the field. This can help to attract funding, partnerships, and help strengthen patent applications related to the custom antibody.

ProteoGenix Can Therapeutically Modify Your Phage Display Antibodies

We can adapt your monoclonal phage display antibody into a bispecific antibody or conjugate it to cytotoxic drugs (ADC antibody) to target diseased tissue. We can also adapt your custom GPCR antibody for diagnostic applications such as ELISA, flow cytometry, or clinical imaging.

Hybridoma Cell Line Generation

Generating your custom anti-GPCR antibody by making a hybridoma cell line is a great way to make anti-GPCR antibodies that bind with high affinity. The first step in producing a hybridoma cell line involves immunizing mice or rats with the GPCR antigen. Next, we collect splenocytes from the immunized mice or rats and fuse the appropriate B-cells with a myeloma cell line.
We select the hybrid cells by screening the supernatant for antibodies that bind the target antigen by ELISA. Lastly, we subject the positive binders to limited dilutions to isolate individual cells and expanded them into colonies. We then screen each colony and further verify their potential to bind antigens by ELISA. The entire process takes 10 weeks from start to finish. Read more about ProteoGenix’s hybridoma technology.

Hybridoma Cell Line Generation Workflow

Antigen procurement or design and production

  • Our experts immunize animal hosts using GPCRs in micelles, nanodiscs, or whole cells that express your GPCR of interest.

Immunization

  • Mice or rats are immunized with purified antigen

Cell Fusion

  • Splenocytes are collected from 2 mice (or rats) to produce 2 fusions from a myeloma cell line.

Hybridoma Selection and Screening (Polyclonal Stage)

  • Hybrid cell selection (HAT selection) culture supernatant screening vs target antigen (ELISA screening).

Isolation of Cell Clones

  • Monoclones are isolated by limited dilution. Expansion and screening of the monoclones by ELISA or in target application.

B-cell Screening

Our B-cell screening process follows a rigorous three-step procedure that ensures the production of monoclonal antibodies with exceptional affinity. To begin, we immunize rodents with purified antigens, which we carefully assess using SDS-PAGE. This process lasts for 6 to 8 weeks and involves our optimized immunization protocol, which consists of 4 to 6 rounds of injections.
The next step involves a 2–3-week process of sorting and screening B-cells, where lymphocytes are isolated from the PBMC compartment and spleen of immunized rodents. These B-cells are then cultured in vitro, and the supernatants are analyzed via ELISA to identify the presence of antibodies that bind to the target antigen. The top-performing antibodies are then sequenced, cloned, and expressed in XtenCHO cells, followed by further ELISA screening to confirm the production of high-quality monoclonal antibodies. Learn more about ProteoGenix’s B-cell screening and isolation methods.

B-cell Screening Workflow

Antigen procurement or design and production

  • Our experts immunize animal hosts using GPCRs in micelles, nanodiscs, or whole cells that express your GPCR of interest.

Immunization

  • Rabbits are immunized using 4-6 rounds of optimized immunizations

FACS Sorting + ELISA screening

  • Antigen-specific B-cell sorting,
    B-cell culture, and supernatant screening by ELISA

Positive Clones Sequenced and Expressed

  • Clones with highest binding affinity are selected and antibodies are transiently expressed using high performance XtenCHO cells

Screening of the antibodies produced