Engineered Upconversion Nanoparticles for Resolving Protein Interactions inside Living Cells

Drees, Christoph; Raj, Abhay; Kurre, Rainer; Busch, Karin B.; Haase, Markus; Piehler, Jacob

doi:10.1002/anie.201603028

Cited by 101 publications

(72 citation statements)

References 72 publications

(19 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Sensitized fluorescence upon LRET from the UCNPs can only be detected when bait and prey interact (in)directly. As proof-of-concept, the indirect interaction between mitochondrial outer membrane (MOM) proteins Tom20 and Tom7 was successfully visualized using UCNP LRET (45). …”

Section: Nanobodies Used As Research Tool To Identify Protein–proteinmentioning

confidence: 99%

“…The nanobody was utilized as delocalization tool, thereby enriching bait and eventually prey at defined subcellular locations (F3H, FRET) (42, 44), or was used to target a reporter toward bait protein (LRET) (45). As such, the GFP nanobody emerges as a highly adaptable research tool to study protein–protein interactions.…”

Section: Nanobodies Used As Research Tool To Identify Protein–proteinmentioning

confidence: 99%

See 1 more Smart Citation

Nanobody Technology: A Versatile Toolkit for Microscopic Imaging, Protein–Protein Interaction Analysis, and Protein Function Exploration

2017

View full text Add to dashboard Cite

Over the last two decades, nanobodies or single-domain antibodies have found their way in research, diagnostics, and therapy. These antigen-binding fragments, derived from Camelid heavy chain only antibodies, possess remarkable characteristics that favor their use over conventional antibodies or fragments thereof, in selected areas of research. In this review, we assess the current status of nanobodies as research tools in diverse aspects of fundamental research. We discuss the use of nanobodies as detection reagents in fluorescence microscopy and focus on recent advances in super-resolution microscopy. Second, application of nanobody technology in investigating protein–protein interactions is reviewed, with emphasis on possible uses in mass spectrometry. Finally, we discuss the potential value of nanobodies in studying protein function, and we focus on their recently reported application in targeted protein degradation. Throughout the review, we highlight state-of-the-art engineering strategies that could expand nanobody versatility and we suggest future applications of the technology in the selected areas of fundamental research.

show abstract

Section: Nanobodies Used As Research Tool To Identify Protein–proteinmentioning

confidence: 99%

Section: Nanobodies Used As Research Tool To Identify Protein–proteinmentioning

confidence: 99%

Nanobody Technology: A Versatile Toolkit for Microscopic Imaging, Protein–Protein Interaction Analysis, and Protein Function Exploration

2017

View full text Add to dashboard Cite

show abstract

“…For nonpolar media, THG emission generated from the volume can be expected to exceed the surface-produced SHG by several orders of magnitude as a result of a higher number of dipoles involved in the former compared to the latter ( f R 1). In the opposite case where χ (2) does not diminish, SHG emission is predicted to be larger than or at least on the order of THG intensity, and therefore f R 1.…”

Section: Introducing a Figure Of Meritmentioning

confidence: 97%

“…The most prominent examples are upconversion nanocrystals [1], where light emission is achieved via sequential absorption to intermediate states [2] featuring sharp emission spectra and long-lived excited states. These exceptional properties have already led to a diversity of applications and a strong focus in the realm of bioapplications such as imaging, sensing, and cancer therapy [3][4][5][6][7][8].…”

Section: Introductionmentioning

confidence: 99%

Nonlinear Diffuse fs-Pulse Reflectometry of Harmonic Upconversion Nanoparticles

et al. 2017

View full text Add to dashboard Cite

Nonlinear diffuse femtosecond-pulse reflectometry is introduced as a powerful experimental tool for the unambiguous characterization of polar and non-polar point symmetry groups of harmonic upconversion nanoparticles. Using intense ultrashort 40 femtosecond laser pulses and an appropriate figure of merit (FOM), second and third harmonic emission serve for the structural characterization of polar Yb-doped lithium niobate and non-polar titanium dioxide nanoparticles. The tool is capable of differentiating these two samples by FOM values that differ by up to 13 orders of magnitude. The general applicability to harmonic upconversion nanoparticles over a broad range of intensities and wavelength spectrum, is discussed.

show abstract

“…Remarkably, TOM7 and TOM20 are spatially separated by TOM40. The capacity of this technique for reporting indirect longdistance interactions might be of interest to unravel cellular protein complexes [85]. …”

mentioning

confidence: 99%

Use, Applications and Mechanisms of Intracellular Actions of Camelid VHHs

Steels¹,

Bertier²,

Gettemans³

2018

Antibody Engineering

View full text Add to dashboard Cite

The discovery of heavy-chain-only antibodies (HCAbs) in camelids and sharks led to the rise of a new research field in which single-domain antibodies are used for various applications. Single-domain antibodies are the antigen-binding fragments derived from HCAbs showing several beneficial properties (e.g., small size, specificity, stability under extreme conditions, cost-effective production, and ease of engineering). Importantly, they are stable in reducing cytoplasmic environment, which allows their use as an intrabody to target a wide range of intracellular targets. In this chapter, we discuss both the therapeutic potential of camelid single-domain antibodies (nanobodies) and their use as a research tool with the main focus on its intracellular employment. Targeting intracellular proteins using nanobodies as a therapeutic per se is, up to now, limited due to its incapacity to traverse the cellular membrane. They can however serve as a stepping stone to small compound development, since they directly target a resident, endogenous protein, similar to how a conventional drug acts. In addition, nanobodies are highly adaptable tools and possess interesting properties for more fundamental research objectives like the elucidation of protein function, the tracking and visualization of endogenous proteins in an in vivo setting, and the assessment of protein-protein interactions.Keywords: VHH, single-domain antibody, nanobody, intrabody, therapy, research tool Nanobodies: a concise introductionIn 1993, Hamers-Casterman discovered the presence of heavy-chain-only antibodies in the sera of Camelidae and assessed that these antibodies are still capable of recognizing an extensive repertoire of antigens despite the absence of the light chain. Single-domain antibodies from camels are called nanobodies. They stated that this discovery could be of inestimable © 2018 The Author(s). Licensee InTech. This chapter is distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/3.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.value to the development and engineering of soluble V H domains or new immunological molecules for diagnostic, therapeutic, and biochemical purposes [1]. This discovery gave rise to a whole new research field in which single-domain antibodies are used for a wide range of applications. Some of these will be reviewed in the current chapter.The structural properties of conventional IgG antibodies are well known. These consist of two heavy-chain polypeptides and two light-chain polypeptides, each of which is folded into four and two domains, respectively. A variable domain is situated at the N-terminus of both chains (VH and VL) and, as the name suggests, its sequence diverges between IgG antibodies. Paired VH-VL domains make up the variable fragment (Fab) and are responsible for the recognition and binding of the target antigen. The sequence of the other domains is well conse...

show abstract

Engineered Upconversion Nanoparticles for Resolving Protein Interactions inside Living Cells

Cited by 101 publications

References 72 publications

Nanobody Technology: A Versatile Toolkit for Microscopic Imaging, Protein–Protein Interaction Analysis, and Protein Function Exploration

Nanobody Technology: A Versatile Toolkit for Microscopic Imaging, Protein–Protein Interaction Analysis, and Protein Function Exploration

Nonlinear Diffuse fs-Pulse Reflectometry of Harmonic Upconversion Nanoparticles

Use, Applications and Mechanisms of Intracellular Actions of Camelid VHHs

Contact Info

Product

Resources

About