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Particle-Mediated DNA Immunization Service

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As a well-known antibody-related service provider, Creative Biolabs is able to provide our worldwide clients the particle-mediated DNA immunization service through the gene gun approach to generate high-quality antibodies against in vivo expressed antigens. Combine with our comprehensive antibody production platform, the particle-mediated DNA immunization method can be employed to a variety of host animals to induce a good immune response with only a small amount of antigen-encoding DNA.

The workflow of a gene gun. Step 1 the gene gun apparatus is ready to fire. Step 2 helium fills the chamber and pressure builds against the rupture disk. Step 3 the pressure eventually reaches the point where the rupture disk breaks, and the resulting burst of helium propels the DNA/gold-coated macrocarrier ('Plastic Disk') into the stopping screen. Step 4 when the macrocarrier hits the stopping screen, the DNA-coated gold particles are propelled through the screen and into the target cells.

DNA Immunization via Gene Gun

The particle-mediated DNA transfer, also refers to gene gun, biolistic particle delivery, biolistics or particle bombardment, is one the commonly used methods contribute to manipulating the gene of organisms. In terms of designed device (the gene gun), the specific “payloads” [metal microparticles (gold or titanium) coated with exogenous DNA/RNAs (e.g. plasmid)] are endowed with high-velocity acceleration to be delivered into the cells of living tissues. This technique has been confirmed the reliability to transform nucleus, organelles (such as plastids), and almost any type of cells. It has now become a versatile method for various genetic engineering researches, disease studies, and a common tool for labeling subsets of cells in the cultured tissue. While for the immunology and antibody production field, gene gun is also a powerful immunization strategy, especially for the delivery of DNA vaccines.

Compared with the classical electroporation and direct intramuscular injection of purified DNA in saline, intradermal immunization via gene gun is a more efficient method, which can induce similar immune response with significantly reduced requirement of immunized plasmid DNA. Particle-mediated DNA immunization is a non-viral transfection method which can simply and efficiently transfect interested DNA/RNAs into the epidermis. Through particle bombardment, keratinocytes (the main cell type in the epidermis) are the predominant transfected cells, while about 5 % blasted targets are Langerhans cells. Langerhans cells are dendritic cells (antigen-presenting cells, APCs) of the skin and mucosa, thus the exogenous proteins (antigens) expressed by the transfect DNAs can be recognized as foreign invaders, which are then processed and presented to alert the immune system and trigger the immune response. Generally, particle-mediated DNA immunization induces Th2-type responses and produces IgG1. Such direct penetration of antigen-presenting cells dramatically increased the interested antigens recognition in the draining lymph nodes resulting in only small amounts of DNA are sufficient to elicit a good immune response.

Antibody Discovery via Gene Gun-mediated DNA Immunization

On the base of gene gun DNA immunization technique, Creative Biolabs can provide a variety of antibody discovery services to our global clients. In addition to the most common polyclonal antibody production, Creative Biolabs can also generate monoclonal antibody with high specificity and affinity via phage display, B-cell sorting or hybridoma technologies. Furthermore, there are a series of host species available at Creative Biolabs to be used for immunization and novel antibody development as well. With years of experience in the antibody discovery realm, our scientists are confident in generating the high-quality antibodies to meet our customers’ specific requirements.

GFP-expressing cells after immunization with plasmid DNA encoding GFP. Left side, expression was observed 24 h after immunization in the mucosa at magnifications of ×540. Right side, expression was observed 24 h after immunization in the dendritic cell in the skin at magnifications of ×270. Fig. 1 GFP-expressing cells after immunization with plasmid DNA encoding GFP. (Loehr et al. 2000)

Comprehensive Antibody Production Platforms of Creative Biolabs


In addition to the gene gun method, DNA immunization services via other techniques are also available at Creative Biolabs, which including but not limited to:


Reference

  1. Loehr, B., Willson, P. and Babiuk, L. (2000) 'Gene gun-mediated DNA immunization primes development of mucosal immunity against bovine herpesvirus 1 in cattle', Journal of virology, 74(13), 6077-6086.



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