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Transforming nanobodies into high-precision tools for protein function analysis.
- Source :
-
American journal of physiology. Cell physiology [Am J Physiol Cell Physiol] 2021 Feb 01; Vol. 320 (2), pp. C195-C215. Date of Electronic Publication: 2020 Dec 02. - Publication Year :
- 2021
-
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.
- Subjects :
- Animals
Antibodies immunology
Antibodies metabolism
Blood-Brain Barrier immunology
Blood-Brain Barrier metabolism
Cell Membrane immunology
Cytoplasm immunology
Humans
Immunotherapy, Adoptive trends
Intracellular Space immunology
Intracellular Space metabolism
Proteins physiology
Single-Domain Antibodies immunology
Cell Membrane metabolism
Cytoplasm metabolism
Immunotherapy, Adoptive methods
Single-Domain Antibodies metabolism
Subjects
Details
- Language :
- English
- ISSN :
- 1522-1563
- Volume :
- 320
- Issue :
- 2
- Database :
- MEDLINE
- Journal :
- American journal of physiology. Cell physiology
- Publication Type :
- Academic Journal
- Accession number :
- 33264078
- Full Text :
- https://doi.org/10.1152/ajpcell.00435.2020