Research Progress and Applications of Multivalent, Multispecific and Modified Nanobodies for Disease Treatment

Abstract

Recombinant antibodies such as nanobodies are progressively demonstrating to be a valid alternative to conventional monoclonal antibodies also for clinical applications. Furthermore, they do not solely represent a substitute for monoclonal antibodies but their unique features allow expanding the applications of biotherapeutics and changes the pattern of disease treatment. Nanobodies possess the double advantage of being small and simple to engineer. This combination has promoted extremely diversified approaches to design nanobody-based constructs suitable for particular applications. Both the format geometry possibilities and the functionalization strategies have been widely explored to provide macromolecules with better efficacy with respect to single nanobodies or their combination. Nanobody multimers and nanobody-derived reagents were developed to image and contrast several cancer diseases and have shown their effectiveness in animal models. Their capacity to block more independent signaling pathways simultaneously is considered a critical advantage to avoid tumor resistance, whereas the mass of these multimeric compounds still remains significantly smaller than that of an IgG, enabling deeper penetration in solid tumors. When applied to CAR-T cell therapy, nanobodies can effectively improve the specificity by targeting multiple epitopes and consequently reduce the side effects. This represents a great potential in treating malignant lymphomas, acute myeloid leukemia, acute lymphoblastic leukemia, multiple myeloma and solid tumors. Apart from cancer treatment, multispecific drugs and imaging reagents built with nanobody blocks have demonstrated their value also for detecting and tackling neurodegenerative, autoimmune, metabolic, and infectious diseases and as antidotes for toxins. In particular, multi-paratopic nanobody-based constructs have been developed recently as drugs for passive immunization against SARS-CoV-2 with the goal of impairing variant survival due to resistance to antibodies targeting single epitopes. Given the enormous research activity in the field, it can be expected that more and more multimeric nanobody molecules will undergo late clinical trials in the next future.Systematic Review Registration

Single-Domain Antibodies as Therapeutics for Respiratory RNA Virus Infections

“…Importantly, these cryptic domains are usually highly conserved and induce a low immune response in humans. Secondly, sdAb can be easily engineered to be multivalent to improve neutralizing potency and breadth [22]. Notably, by inhaled delivery, sdAbs can be directly delivered to the lung as the main infectious tissue in respiratory diseases [25].…”

“…In addition, sdAbs can be easily engineered to be multivalent or multispecific, improving their binding affinity and breath [22] and prolonging half-life in vivo [23]. Lauren et al reported four sdAbs, designed as SD36, SD38, SD83, and SD84, which could bind to highly conserved epitopes of the influenza A and B virus hemagglutinins (HAs), respectively.…”

Isolation of SARS-CoV-2-blocking recombinant antibody fragments and characterisation of their binding to variant spike proteins

“…In addition, the ability of antibodies to block or outcompete specific binding interactions Frontiers in Nanotechnology frontiersin.org has led to their use in neutralising viruses and toxins (Cheedarla and Hanna, 2019). As the classical hybridoma technology developed with murine B cells (Köhler and Milstein, 1975) has proven difficult to adapt to the generation of human monoclonal antibodies, however, recombinant approaches have been widely developed to create human antibodies, or derived antibody fragments (Wronska et al, 2016), which can be used in vivo in applications such as drug delivery and neutralisation, and in vitro methodologies like immunodiagnosis (Raeisi et al, 2022;Wang et al, 2022).…”

Design of nanobody-based bispecific constructs by in silico affinity maturation and umbrella sampling simulations

“…In this perspective, the development of bispecific antibodies targeting simultaneously both antigens might be considered a logic development, similarly to what already positively tested in the case of other diseases [17] . Since such dual-activity molecules are technically difficult to prepare with IgG, recombinant antibody fragments represent a convenient alternative to produce multi-specific binders the realization of which requires extensive engineering [18] . The availability of a bifunctional molecule, instead of two independent binders, might also simplify the drug characterization since it will halve the work and avoid dealing with unpredictable interferences between the two single molecules.…”