Hybridoma technology a versatile method for isolation of monoclonal antibodies, its applicability across species, limitations, advancement and future perspectives

Abstract

The advancements in technology and manufacturing processes have allowed the development of new derivatives, biosimilar or advanced improved versions for approved antibodies each year for treatment regimen. There are more than 700 antibody-based molecules that are in different stages of phase I/II/ III clinical trials targeting new unique targets. To date, approximately more than 80 monoclonal antibodies (mAbs) have been approved. A total of 7 novel antibody therapeutics had been granted the first approval either in the United States or European Union in the year 2019, representing approximately 20% of the total number of approved drugs. Most of these licenced mAbs or their derivatives are either of hybridoma origin or their improvised engineered versions. Even with the recent development of high throughput mAb generation technologies, hybridoma is the most favoured method due to its indigenous nature to preserve natural cognate antibody pairing information and preserves innate functions of immune cells. The recent advent of antibody engineering technology has superseded the species level barriers and has shown success in isolation of hybridoma across phylogenetically distinct species. This has led to the isolation of monoclonal antibodies against human targets that are conserved and non-immunogenic in the rodent. In this review, we have discussed in detail about hybridoma technology, its expansion towards different animal species, the importance of antibodies isolated from different animal sources that are useful in biological applications, advantages, and limitations. This review also summarizes the challenges and recent progress associated with hybridoma development, and how it has been overcome in these years to provide new insights for the isolation of mAbs.

Immunomodulatory Potential of anti-idiotypic Antibodies for the Treatment of Autoimmune Diseases

“…Generally, these antibodies do not rely on external antigen or fragments, which is the bottleneck factor in the production of conventional vaccines [ 49 ]. Monoclonal antibodies can be produced from hybridoma-based technologies which can recognise a single epitope of an antigen [ 94 ]. For example, in 1995, Foon et al successfully generated a murine monoclonal anti-ID antibody which mimics a specific epitope on a tumour-associated antigen known as the carcinoembryogenic antigen using hybridoma-based technology [ 95 ].…”

Alternative Methods to Animal Use for Monoclonal Antibody Generation and Production

“…This methodology is based on the immunization of animals with the antigen of interest, followed by the fusion of B lymphocytes with myeloma cells, resulting in the formation of hybridomas. After cloning and selection, the antibody-secreting stable monoclonal cell lines were used to produce mAb-enriched ascitic liquid [8], a now-banned practice. For many years, mAbs, which have wide applicability in research, diagnosis, and treatment, were generated and produced with this methodology.…”

Directed evolution of conformation‐specific antibodies for sensitive detection of polypeptide aggregates in therapeutic drug formulations

“…Antibody‐based detection assays are well known to be highly sensitive and are attractive for detecting extremely low levels of polypeptide aggregates. However, immunization‐based generation of conformation‐specific antibodies against polypeptide aggregates is challenging for many reasons, including difficulties in controlling antigen presentation to the immune system, weakly immunogenic antigens in some cases and immunodominant epitopes in other cases, and the fact that most antibodies generated via this process recognize linear (nonconformational) epitopes (Kayed & Glabe, 2006; Lambert et al, 2009; Parray et al, 2020; Vaikath et al, 2019). To address these common challenges, we have used directed evolution methods to generate large antibody libraries and sorted them to isolate conformation‐specific antibodies against polypeptide aggregates of a therapeutic GLP‐1 agonist (liraglutide) used for treating diabetes and obesity (Croom & McCormack, 2009; Drucker et al, 2010).…”