Next-Generation Techniques for Discovering Human Monoclonal Antibodies

Abstract

Abstract⎯Monoclonal antibodies have found wide applications in the treatment of cancer, as well as of autoimmune, infectious, and other diseases. Several dozen new antibodies are currently undergoing different stages of clinical trials, and some of them will soon be added to the list of immunotherapeutic drugs. Most of these antibodies have been generated using hybridoma technology or a phage display. In recent years, new methods of obtaining human monoclonal antibodies have been actively developing. These methods rely on sequencing immunoglobulin genes from B lymphocytes, as well as on the creation of antibody-secreting stable B-cell lines. The term next-generation antibody-discovery platforms has already been established in the literature to refer to these approaches. Our review focuses on describing the results obtained by these methods.

Considerations for the Design of Antibody-Based Therapeutics

“…118,123 As hybridoma technology functions in the context of an intact immune system, only certain antigens can be effectively targeted using this method. 124 A primary consideration is that antigens must be sufficiently immunogenic to elicit a humoral response in the host animal; thus, antibodies targeting endogenous proteins may be difficult to generate in vivo. On the other hand, it is often not feasible or ethical to inject mice with pathogens or toxins that may prove fatal before the generation of a robust B-cell response.…”

“…B-cell receptor sequencing (BCR seq) uses next-generation sequencing to read the variable region sequences of up to several million Ig variants. 124 This method is useful not only for the antibody discovery but also for detailed characterization of humoral immune responses. 125 While the peripheral blood compartment is the most accessible source of human B cells, most B cells are found in extravascular milieu such as the spleen, bone marrow, and lymph nodes and can only be accessed in laboratory animals.…”

“…126 The localization and time since antigen exposure also affect the functional phenotype of B cells; for example, plasmablasts are abundant in blood 1 week after antigen exposure, while memory B cells appear 1 to 2 weeks later. 124 The B-cell subset, in turn, affects properties such as messenger RNA (mRNA) levels and extent of affinity maturation. Thus, the choice to use mRNA or genomic DNA as the template for BCR seq depends whether information on mRNA expression or Bcell clonality is desired.…”

“…125 This method is especially powerful after immunization, when Ag-specific B cells make up a significant portion of the total B-cell pool. 124,126 For antibodies that have undergone significant somatic hypermutation, pairing based on similar levels of mutation accumulation is another approach that has produced viable antibodies. 126 The most reliable way to ensure physiological pairing of variable sequences, however, is to clone individual BCRs.…”

<p>The importance of early identification of infusion-related reactions to monoclonal antibodies</p>

“…Important recent advances in biomedical science have been associated with the development and clinical application of biological agents based on monoclonal antibodies (mAb) 1. These have been used as important tools in clinical practice and biotechnology in diagnosing and treating infectious, inflammatory, immunological and neoplastic diseases, and also for examining the host/pathogen interaction, and for detecting and quantifying diverse molecules 2…”