DNA immunization services and technologies have been developed and optimized at ProteoGenix in order to offer a successful alternative for custom polyclonal or monoclonal antibody generation against difficult-to-express targets but also for early DNA vaccine development studies. There are several other strong advantages of genetic immunization for antibody production over other antibody development methods.

Hybridoma development via genetic immunization Polyclonal antibody generation by genetic immunization Immune antibody library via DNA immunization
  • Codon optimization
  • Gene Synthesis
  • Plasmid preparation
  • Animal injections
  • Fusion and subcloning
  • Antibody production
  • Codon optimization
  • Gene Synthesis
  • Plasmid preparation
  • Animal injections
  • Serum collection
  • Antibody purification
  • Codon optimization
  • Gene Synthesis
  • Plasmid preparation
  • Animal injections
  • Cells Isolation & RNA extraction
  • DNA synthesis
  • VH-VL amplification & cloning
  • Phage library creation of at least 108
  • QC / sequencing
Strating Material Antigen sequence Antigen sequence Antigen sequence
Deliverable Culture supernatants Protein A/G purified serum Phage library
Lead Time 3 to 6 months 2 to 3 months 3 to 5 months
Starting from 5 800 840 25 500

Services comparisons – DNA Immunization advantages

  • Native protein: Antigen shows proper Post Translational Modifications (PTM) that triggers conformational epitopes leading to higher affinity antibodies, crucial for many applications and especially most therapeutic antibody developments.
  • Save time: No need for antigen production.
  • Cost less: No need to produce and purify large amounts of antigen.
  • No contaminant injected: Co purified proteins, protein purification tags, HCP
  • High antibody titer: Smaller injections and steady presentation to immune system favors high titers and therefore higher affinity antibodies.
  • Applications: Antibodies will work in a broader range of application as the antigen presents linear epitopes but also conformational epitopes.
  • No Antigen needed: Antigens are sometimes difficult to express or purify and to find in catalogs because they are toxic, insoluble, unstable or membrane bound. When produced in recombinant way, it can be different from the native form especially depending on the protein expression system used and therefore lead to irrelevant antibodies.

ProteoGenix’ team of account managers is available to study each customer project in order to define the most suitable strategy to develop antibodies meeting customers’ specifications in terms of antibody performances, deliverables, budget and lead time. Our wide range of technologies and solutions allows us to design optimal strategies no matter the constraints we have to face.

For DNA immunization programs more specifically, we possess an unrivaled experience in all steps of the process including codon optimization, gene synthesis, plasmid preparation, animal injection, hybridoma development and antibody purification. This experience allows us to provide integrated antibody production services by genetic immunization delivering antibodies of strong affinity at very affordable prices in a short period of time.

How does our genetic immunization service work?

What material are antibodies screened and selected on?

  • Either you have very small amounts of purified protein or we can transfect cells with the plasmid and test the antibodies on these transfected cells directly or protein produced and purified from these cells. We can also perform control screenings on negative cell lines or antigens which shouldn’t be detected.

Is this methos suited for all types of antigen species?

  • DNA immunization is perfectly suited for proteins from mammalian origin, but not for prokaryotic proteins. Indeed, animal in situ expression of prokaryotic antigens may lead to wrong post translational modifications creating a protein structure far different from the wild type prokaryotic version. So we recommend producing a recombinant protein for prokaryotic antigens for better success rates. All eukaryotic species other than mammalian species should be assessed case by case to define the best immunization strategy between DNA immunization and recombinant protein immunization.

In which cell type is the antigen transfected?

  • It strongly depends on the injection site. For instance, hydrodynamic tail vein (HTV) delivery predominantly triggers transfection in hepatocytes (but not in Kupffer cells) and in smaller amounts in the spleen, lung, and cardiac muscle whereas hydrodynamic limb vein (HLV) delivery predominantly triggers DNA transfection in skeletal muscles.

Who should prepare the plasmid to inject?

  • To make sure to use the best cDNA, vector and the best preparation for injection, we strongly recommend letting ProteoGenix take care of it. As a leader in gene synthesis, our prices are among the most competitive on the market and our services among the fastest of the industry. Integrating all steps of the service provides you the best guarantees of success, the fastest and most efficient way to complete the service, avoids unnecessary logistic costs and reduces unnecessary inputs between you and the lab.

Are nuclear antigens and transmembrane antigens good targets?

  • Nuclear antigens are not suitable as the produced protein may stay predominantly in the nucleus and will not be made available to the immune system unless we use some tricks to try to secrete it.
    However, there is no problem with transmembrane proteins as they will trigger an immune response.

Genetic immunization services applications

DNA immunization principle

The principle of genetic immunization for antibody production is to inject to animals a plasmid DNA coding for the targeted antigen instead of injecting the target itself. Our proprietary plasmid that also serves as an adjuvant will transfect animal cells and conduct to in situ expression of the antigen, that will therefore be presented to the immune system in a form usually much closer to the native form of that antigen. The animal then responds normally to the protein target it has expressed in vivo. DNA immunization is therefore an outstanding tool to develop polyclonal and monoclonal antibodies against certain types of targets or to study DNA vaccine candidates at early stage. After years of research ProteoGenix has established high performing SOPs to develop high affinity antibodies through genetic immunization services.