DNA immunization as a technology platform for monoclonal antibody induction

Abstract

To combat the threat of many emerging infectious diseases, DNA immunization offers a unique and powerful approach to the production of high-quality monoclonal antibodies (mAbs) against various pathogens. Compared with traditional protein-based immunization approaches, DNA immunization is efficient for testing novel immunogen designs, does not require the production or purification of proteins from a pathogen or the use of recombinant protein technology and is effective at generating mAbs against conformation-sensitive targets. Although significant progress in the use of DNA immunization to generate mAbs has been made over the last two decades, the literature does not contain an updated summary of this experience. The current review provides a comprehensive analysis of the literature, including our own work, describing the use of DNA immunization to produce highly functional mAbs, in particular, those against emerging infectious diseases. Critical factors such as immunogen design, delivery approach, immunization schedule, use of immune modulators and the role of final boost immunization are discussed in detail.

Antifungal Susceptibility Testing of Fusarium: A Practical Approach

“…Delayed initiation of antifungal therapy due to non-optimal diagnosis of mold infections are associated with increased mortality rates and may lead to excessive drug use in prophylaxis and therapy [ 1 , 6 ]. The Fusarium solani species complex contains many opportunistic species (e.g., F. falciforme , F. keratoplasticum and F. petroliphilum ) with high prevalence, but other Fusarium groups are also important, such as F. oxysporum , F. verticillioides and F. proliferatum , and susceptibility to antifungal agents varies between species [ 7 ]. Treatment strategies for fusariosis have been evaluated [ 8 ] which led to the European Society of Clinical Microbiology and Infectious Diseases (ESCMID) ESCMID and European Confederation of Medical Mycology (ECMM) guidelines for management of hyalohyphomycosis caused by Fusarium and other non-melanized fungi [ 9 ].…”

A review of antibody-based therapeutics targeting G protein-coupled receptors: an update

“…Recent developments designed to enhance the ability for antibody discovery and functional screening include increasing cell surface expression and methods for detection of GPCRexpressing cell lines [21,22]; access to new generation detergents such as calixarenes [23], GPCR stabilization for improved expression [24,25], enhanced thermostability or conformation [26], the implementation of lipoparticles [27], magnetic proteoliposomes [28] or virus-like particles [29], spherical-supported bilayer lipid membranes [30] and the generation of nanodiscs [31] have all added to the arsenal of antigen formats that can be applied not only in the generation of antibodies but also associated downstream discovery processes and compliment established tool reagents, such as whole cells or membranes over-expressing the target of interest. The effectiveness of DNA immunization expression constructs or GPCR-expressing cells can be enhanced using adjuvants [32,33] with inventive selection strategies [34] and affinity maturation cell-based approaches devised, for example, CHO cell display libraries of single-chain variable fragments The typical structure for each major GPCR family is presented where the blue horizontal lines represent the lipid bilayer of the cell membrane and numbers 1-7 each represents a GPCR helix or individual transmembrane (TM) domain. An example of each family member is provided alongside the respective endogenous ligand(s), e.g., in the case of CCR5, there are three main chemokine ligands.…”

Camelid Single-Domain Antibodies: Historical Perspective and Future Outlook

“…Immunization of large animals and heterogeneity in immune responses among individual outbred animals is another consideration which is important when alternative immunization techniques such as DNA immunization are applied. DNA immunization has had limited success in camelid and other large animals and reproducibility is often a major issue to be tackled ( 84 – 87 ). To overcome this limitation, transgenic mice bearing either a rearranged dromedary γ2a chain or hybrid llama/human antibody loci have been generated that produce a form of dromedary or human heavy chain antibodies ( 88 – 90 ).…”