DNA Immunization Introduction for Hybridoma Generation

Compared with traditional protein-based immunization strategies, DNA immunization does not require the production or purification of proteins from pathogens or the use of recombinant protein technology, and it can effectively generate monoclonal antibodies against conformational sensitive targets. Therefore, DNA immunization is effective for testing novel immunogen designs.

Optimal Design of Immunogen Inserts

If the protein is naturally expressed in membrane-associated forms, such as multitransmembrane G protein-coupled receptors (GPCRs) and ion channels, it is difficult to generate full-length protein immunogens by recombinant protein approaches using traditional immunization methods. However, the intact protein structure is essential for the induction of functional monoclonal antibodies (mAbs). Expression of the intact immunogen in vivo by DNA immunization seems most likely to induce mAbs with the desired biological activity. On the one hand, immunogen inserts expressing full-length sequences of target proteins are commonly used with good success for all types of proteins, especially transmembrane proteins. On the other hand, the gene sequence of the DNA vaccine immunogen insert can be easily edited to express the "designer proteins".

Delivery Approach

At present, there are two main ways to deliver plasmids obtained. One is to dissolve the DNA plasmid in various solutions for conventional needle injection. The solution can include additional facilitating agents, such as lipids and nanoparticles, to improve delivery efficacy. The second is delivery based on physical forces. Among them, the most representative method is the use of gene guns, which use "ballistic" forces to deliver DNA plasmids. Another physical method is the use of electroporation. In general, gene gun and electroporation delivery are more effective in eliciting higher antibody response levels than conventional intramuscular needle injections. In addition, in contrast to electroporation, the gene gun method requires only a few micrograms of DNA plasmid to achieve the same level of immune response elicited by electroporation (over 100 μg).

After the DNA immunization process is complete, the immune plasmid can be transferred into the cell line using DNA transfection reagents (such as DNA transfection reagents, mRNA transfection reagents, and siRNA transfection reagents) to verify the immune response and further screening.

More Details on DNA Immunization for Hybridoma Generation

Introduction to Gene Gun DNA Immunization for Hybridoma Generation
Introduction to Electroporation DNA Immunization for Hybridoma Generation
Introduction to Naked DNA Immunization for Hybridoma Generation
Introduction to Heterologous Immunization for Hybridoma Generation

DNA immunization provides a unique and effective approach for the production of high-quality monoclonal antibodies against a variety of pathogens to deal with the threat of many emerging infectious diseases. Creative Biolabs is willing to share our expertise and experience on DNA immunization to help you successfully complete your research project.

Reference

Mitra, S. Tomar, P. C., Hybridoma technology; advancements, clinical significance, and future aspects. J Genet Eng Biotechnol. 2021. 19(1): p. 159.

All listed services and products are For Research Use Only. Do Not use in any diagnostic or therapeutic applications.

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