Are you struggling to find a source of custom monoclonal antibodies that selectively bind MHC-peptide complexes? ProteoGenix has the solution. Our team of antibody experts can make your anti-MHC-peptide complex antibody using a range of animal species and services to fit your needs, including custom phage display libraries, single B cell clones, or hybridoma cell lines. Whatever your antibody needs, ProteoGenix will combine its resources, services, and 20 years of expertise to deliver reliable anti-MHC-peptide complex antibodies that consistently perform.

Hybridoma Phage Display B-cell Screening
Species Mouse/Rat No Limitations Rabbit/Humanized Mice (Coming Soon)
Human antibodies No Yes No
Direct access to sequences No Yes No
Potential antigen immunogenicity Yes No (Naïve Library Only) Yes
Potential clone viability issue Yes No Yes
Antibody Sensitivity High Medium High
Budget + +++ ++
Lead Time +++ + ++

Antibody Phage Display

Antibody phage display involves collecting peripheral blood mononuclear cells (PBMCs) from immunized animal hosts (immune libraries) or unimmunized hosts (naïve libraries). Once PBMCs are collected, the antibody gene segments are converted into cDNA and cloned into a (bacteriophage equivalent of a plasmid) phagemid to make an antibody-phage fusion protein, exposing the antibody to the outer surface of the bacteriophage.

The target antigen is cloned and immobilized so it can bind the target antibody, a process called biopanning. Once bound, the DNA inside the bacteriophage (containing the antibody-related genes) is isolated, sequenced, and expressed to verify the antibody binds antigen with high affinity. Learn more about how using phage display library construction in less than 8 weeks.

Naive Library Workflow

Antigen procurement or design and production

  • Antigen provided by the customer
  • Or antigen design and production by ProteoGenix

Library screening and biopanning

  • Screening of naive or immune library against antigen.
  • 4-6 rounds of biopanning

ELISA screening of single phage binders

  • ELISA screening and validation until identification of at least 3-10 different binders.

DNA extraction & antibody sequencing

Immune Library Workflow

Antigen procurement or design and production

  • Antigen provided by the customer
  • Or antigen design and production by ProteoGenix

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 against antigen.
  • 4-6 rounds of biopanning

ELISA screening of single phage binders

  • ELISA screening and validation until identification of at least 3-10 different binders.

DNA extraction & antibody sequencing

Why Antibody Phage Display is Ideal for Making Human Therapeutic Antibodies

Antibody phage display is useful for generating monoclonal antibodies to target antigens that traditionally evade most antibody production methods. This includes cancer neoantigens, (such as MHC-peptide complexes), conserved antigens, and human antigens of critical importance. Therefore, if your biomedical research lab or pharmaceutical company is interested in fast-tracking human immunotherapy projects to treat cancer, autoimmunity, or other diseases, then antibody phage display technology is the appropriate method.

Phage Display Has No Antibody Species Limitations

Phage display technology has no species limitations. This means you can engineer custom phage display libraries in species ranging from humans to llamas. However, if you are interested in antibodies for clinical use, bypass the time-consuming process of antibody humanization by making your custom naïve library from human PBMCs. This convenience factor makes antibody phage display technology the preferred method to generate immunotherapeutic or diagnostic antibodies.

Phage Display Deliverables are Antibody DNA Sequences, Not Fragile Cell Lines

The other advantage antibody phage display offers over hybridoma and B-cell screening is the benefit of not having to keep fragile cell lines healthy and viable.

There are several ways labs can lose antibody-producing cell lines. The first is from Mycoplasma contamination. Mycoplasma is a slow-growing intracellular bacterium resistant to antibiotics found in cell medium. In fact, 15% to 35% of all cell lines tested are positive for Mycoplasma contamination. This type of biological contamination is notoriously difficult to treat, often destroying the entire cell stock.

Human error is another common reason laboratories lose cell lines. Stocks of cells must be stored long-term in liquid nitrogen. If frozen cell line stocks thaw because of a liquid nitrogen refill mistake then your precious stock of antibody-producing cells is gone.

Phage display overcomes these limitations because fragile cell clones are never produced. Instead, we identify three antibodies that bind your target anti-MHC-peptide complex and deliver the antibody sequences to you. The best part? You retain ownership of the antibody sequences. They are your intellectual property. Thus, you can modify the antibodies for therapeutic purposes or allow ProteoGenix to modify the antibodies for you.

ProteoGenix’s Pre-Built Human Phage Display Libraries

Convert your bench-side discovery into a life-saving bedside therapy using our pre-built human cancer or autoimmunity phage display libraries. ProteoGenix built the world’s first human cancer and human autoimmune phage display libraries to supercharge your immunotherapeutic projects, saving you time, money, and effort.

Our Pre-built Antibody Phage Display Library for use in Cancer Immunotherapy

The process of a healthy cell transforming into a cancer cell is associated with genetic instability that results in the expression of new antigens, termed neoantigens. These cancer neoantigens are presented by MHC-I causing T-cell activation and cancer cell death. However, when a cancer cell evades T-cell detection, uncontrolled cell division occurs triggering the formation if a malignant tumor.

Adaptive antibody responses are mounted against cancer neoantigens, including the cancer cell peptides presented by MHC I. The antibodies that selectively recognize MHC I-cancer peptide complexes are termed TCR-like antibodies.

TCR-like antibodies hold promising immunotherapeutic potential because they selectively bind cancer cells. Therefore, ProteoGenix captured the antibody repertoire from cancer patients in the form of an antibody phage display library to help scientists accelerate their therapeutic and diagnostic cancer projects (read more about TCR-like antibodies below). Therefore, if you discovered a novel MHC-antigen complex, ProteoGenix can generate your clinical monoclonal antibodies from start to finish using our pre-built human cancer library.

Our Pre-built Antibody Phage Display Library for use in Autoimmune Immunotherapy

One autoimmune disease risk factor is the expression of specific MHC II “risk” alleles by autoreactive antigen-presenting cells (autoAPCs). These MHC II alleles allow the escape of self-tolerance by inadvertently stimulating T-cells when presenting self-antigens.

Although rare, the adaptive immune system can recognize autoantigen presented by MHC II as foreign and build an antibody repertoire to target the MHC II-autoantigen complexes. As in cancer treatment, these TCR-like antibodies have the potential to selectively target and destroy harmful cells including autoreactive APCs.

Therefore, ProteoGenix developed the world’s first naïve human autoimmunity phage display library to help scientists accelerate the discovery of novel anti-MHC-autoantigen complexes for therapeutic and diagnostic applications. Once we identify monoclonal antibody clones that bind your MHC II-autoantigen complex, ProteoGenix can convert it into an antibody-drug complex antibody (ADC) or a bispecific antibody. Therefore, if you discovered a novel MHC-autoantigen complex, ProteoGenix can generate immunotherapeutic monoclonal antibodies from start to finish using our human autoimmune library.

Non-human Antibody Phage Display Libraries

ProteoGenix also offers non-human pre-built naïve antibody phage display libraries. If you are interested in making a custom anti-MHC-peptide complex antibody targeting a non-human MHC then choose from one of the following libraries:

  • Rabbit naive antibody phage display library
  • Camelid naïve antibody phage display library
  • Dog naïve antibody phage display library.

Don’t see a prebuilt library compatible with your research?
Let us make your custom library. Custom libraries give you unlimited access to an array of antibodies that target different MHC-peptide epitopes. This service is perfect for those who know they will need a constant source of antibodies to target yet-to-be-identified MHC-peptide complexes in their disease model.

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 anti-MHC-peptide complex antibody for diagnostic applications such as ELISA, flow cytometry, or clinical imaging.

Hybridoma Cell Line Generation

Generating a hybridoma cell line is a great way to make anti-MHC antibodies that bind MHC-peptide complexes with high affinity. The first step in producing a hybridoma cell line involves immunizing mice or rats with the purified target 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 antigen 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

  • Antigen provided by the customer
  • Or antigen design and production by ProteoGenix

Immunization

  • Mice are immunized with purified antigen

Cell Fusion

  • Splenocytes are collected from 2 mice 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

B-cell screening is a three-step process that produces high-affinity monoclonal antibodies. The first step is a 6- to 8-week process that starts with immunizing rodents with purified antigen assessed by SDS-PAGE. The immunization process has 4-6 rounds of injections using an optimized immunization protocol.

The second step is a 2-3 weeks process involving B-cell sorting and screening. This is achieved by isolating lymphocytes from the PBMC compartment and the spleen of immunized rodents. Lastly, the resulting B-cells are cultured in vitro and the supernatants are assessed for the presence of antibodies that bind the target antigen by ELISA. The resulting best positive antibodies are sequenced cloned, and expressed in XtenCHO cells. Antibody supernatants are further screened by ELISA to confirm the presence 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

  • Antigen provided by the customer
  • Or antigen design and production by ProteoGenix

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

The Challenges of Making an Anti-MHC Peptide Complex Antibody

Designing monoclonal antibodies that selectively bind MHC-peptide complexes, known as TCR-like antibodies, is a difficult task that few companies attempt. However, our antibody experts use their 25 years of average experience to design and troubleshoot the most daunting antibody eng