RRC ID 70849
Author Gettemans J, De Dobbelaer B.
Title Transforming nanobodies into high-precision tools for protein function analysis.
Journal Am J Physiol Cell Physiol
Abstract Single-domain antibodies, derived from camelid heavy antibodies (nanobodies) or shark variable new antigen receptors, have attracted increasing attention in recent years due to their extremely versatile nature and the opportunities they offer for downstream modification. Discovered more than three decades ago, these 120-amino acid (∼15-kDa) antibody fragments are known to bind their target with high specificity and affinity. Key features of nanobodies that make them very attractive include their single-domain nature, small size, and affordable high-level expression in prokaryotes, and their cDNAs are routinely obtained in the process of their isolation. This facilitates and stimulates new experimental approaches. Hence, it allows researchers to formulate new answers to complex biomedical questions. Through elementary PCR-based technologies and chemical modification strategies, their primary structure can be altered almost at leisure while retaining their specificity and biological activity, transforming them into highly tailored tools that meet the increasing demands of current-day biomedical research. In this review, various aspects of camelid nanobodies are expounded, including intracellular delivery in recombinant format for manipulation of, i.e., cytoplasmic targets, their derivatization to improve nanobody orientation as a capturing device, approaches to reversibly bind their target, their potential as protein-silencing devices in cells, the development of strategies to transfer nanobodies through the blood-brain barrier and their application in CAR-T experimentation. We also discuss some of their disadvantages and conclude with future prospects.
Volume 320(2)
Pages C195-C215
Published 2021-2-1
DOI 10.1152/ajpcell.00435.2020
PMID 33264078
MeSH Animals Antibodies / immunology Antibodies / metabolism Blood-Brain Barrier / immunology Blood-Brain Barrier / metabolism Cell Membrane / immunology Cell Membrane / metabolism* Cytoplasm / immunology Cytoplasm / metabolism* Humans Immunotherapy, Adoptive / methods* Immunotherapy, Adoptive / trends Intracellular Space / immunology Intracellular Space / metabolism Proteins / physiology Single-Domain Antibodies / immunology Single-Domain Antibodies / metabolism*
IF 3.485
Resource
Drosophila