Single-Domain Antibodies or Nanobodies: A Class of Next-Generation Antibodies

Khodabakhsh, Farnaz; Behdani, Mahdi; Rami, Abbas; Kazemi‐Lomedasht, Fatemeh

doi:10.1080/08830185.2018.1526932

Cited by 89 publications

(50 citation statements)

References 45 publications

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“…To visualize endogenous proteins in living cells, FP‐tagged nanobodies (chromobodies) provide several advantages. Nanobodies are the smallest antibody structure with binding capacity . Chromobodies are expressed as FP‐targeting endogenous epitopes compatible with live‐cell imaging and super‐resolution approaches .…”

Section: General Considerations When Imaging Admentioning

confidence: 99%

Imaging the adenovirus infection cycle

Pied

Wodrich

2019

FEBS Letters

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Edited by Urs GreberIncoming adenoviruses seize control of cytosolic transport mechanisms to relocate their genome from the cell periphery to specialized sites in the nucleoplasm. The nucleus is the site for viral gene expression, genome replication, and the production of progeny for the next round of infection. By taking control of the cell, adenoviruses also suppress cell-autonomous immunity responses. To succeed in their production cycle, adenoviruses rely on wellcoordinated steps, facilitated by interactions between viral proteins and cellular factors. Interactions between virus and host can impose remarkable morphological changes in the infected cell. Imaging adenoviruses has tremendously influenced how we delineate individual steps in the viral life cycle, because it allowed the development of specific optical markers to label these morphological changes in space and time. As technology advances, innovative imaging techniques and novel tools for specimen labeling keep uncovering previously unseen facets of adenovirus biology emphasizing why imaging adenoviruses is as attractive today as it was in the past. This review will summarize past achievements and present developments in adenovirus imaging centered on fluorescence microscopy approaches.Adenoviruses (Ads) are nonenveloped icosahedral viruses with a diameter of~90 nm with slight structural differences between genotypes. The majority of the Ad capsid shell is composed of 240 hexon trimers, and each of the 12 vertices of the icosahedron is occupied by a penton. Pentons are involved in cell attachment and receptor recognition and are composed of the pentameric penton base from which the trimeric fiber molecule extends. Minor capsid proteins IIIa, VI, VIII, and IX are embedded in the capsid and contribute to capsid stability. Protein VI is located at the inner surface of the capsid, and biochemical data suggest that it may connect the capsid to the core containing the viral genome via protein V. The genome is ã 36-kb double-stranded linear DNA molecule with the terminal protein (TP) covalently bound to each 5 0end. Adenovirus genomes are highly condensed and organized into chromatin by several hundred copies of Abbreviations ADP, adenovirus death protein

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Section: General Considerations When Imaging Admentioning

confidence: 99%

Imaging the adenovirus infection cycle

Pied

Wodrich

2019

FEBS Letters

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“…Nanobodies are VHHs from the camelid family, which lacks the light chain. Currently, a variety of nanobodies have entered the clinical research stage (35)(36)(37). Compared with traditional antibodies, nanobodies have the advantages of low MW, high affinity, high stability, low immunogenicity, and strong penetrability.…”

Section: Discussionmentioning

confidence: 99%

Targeting RyR2 with a phosphorylation site–specific nanobody reverses dysfunction of failing cardiomyocytes in rats

et al. 2019

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Chronic PKA phosphorylation of ryanodine receptor 2 (RyR2) has been shown to increase diastolic sarcoplasmic reticulum (SR) Ca2+ leakage and lead to cardiac dysfunction. We hypothesize that intracellular gene delivery of an RyR2‐targeting phosphorylation site—specific nanobody could preserve the contractility of the failing myocardium. In the present study, we acquired RyR2‐specific nanobodies from a phage display library that were variable domains of Camelidae heavy chain—only antibodies. One of the nanobodies, AR185, inhibited RyR2 phosphorylation in vitro and was chosen for further investigation. We investigated the potential of adeno‐associated virus (AAV)9—mediated cardiac expression of AR185 to combat postischemic heart failure (HF). AAV gene delivery elevated the intracellular expression of the AR185 protein in a rat model of ischemic HF, and this treatment normalized the systolic and diastolic dysfunction of the failing myocardium in vivo by reversing myocardial Ca2+ handling. Furthermore, AR185 gene transfer to failing cardiomyocytes reduced the frequency of SR calcium leaks, thereby restoring the attenuated intracellular calcium transients and SR calcium load. Moreover, AR185 gene transfer inhibited the PKA‐mediated phosphorylation of RyR2 in failing cardiomyocytes. Our results provide preclinical experimental evidence that the cardiac expression of RyR2 nanobodies with AAV9 vectors is a promising therapeutic strategy for HF.—Li, T., Shen, Y., Lin, F., Fu, W., Liu, S., Wang, C, Liang, J., Fan, X., Ye, X., Tang, Y., Ding, M., Yang, Y., Lei, C., Hu, S. Targeting RyR2 with a phosphorylation site–specific nanobody reverses dysfunction of failing cardiomyocytes in rats. FASEB J. 33, 7467–7478 (2019). http://www.fasebj.org

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“…Biotherapeutics, in particular, therapeutic antibodies, provide an important mode to target a number of disease states, and have become well accepted in the market place (Cruz & Kayser, ; Khodabakhsh, Behdani, Rami, & Kazemi‐Lomedasht, ). Importantly, monoclonal antibodies (mAbs) are excellent drug candidates due to their high antigen specificity, favorable PK properties compared with small molecules, and engineering‐friendly modular structure (Chames, Van Regenmortel, Weiss, & Baty, ; Reichert, ).…”

Section: Introductionmentioning

confidence: 99%

Validation of a de‐immunization strategy for monoclonal antibodies using cynomolgus macaque as a surrogate for human

Kovalova

Boyles

Wen

et al. 2020

Biopharm & Drug Disp

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The immunogenicity of biotherapeutics presents a major challenge during the clinical development of new protein drugs including monoclonal antibodies. To address this, multiple humanization and de-immunization techniques that employ in silico algorithms and in vitro test systems have been proposed and implemented. However, the success of these approaches has been variable and to date, the ability of these techniques to predict immunogenicity has not been systematically tested in humans or other primates. This study tested whether antibody humanization and deimmunization strategies reduce the risk of anti-drug antibody (ADA) development using cynomolgus macaque as a surrogate for human. First human-cyno chimeric antibodies were constructed by grafting the variable domains of the adalimumab and golimumab monoclonal antibodies onto cynomolgus macaque IgG1 and Igκ constant domains followed by framework germlining to cyno to reduce the xenogenic content.Next, B and T cell epitopes and aggregation-prone regions were identified using common in silico methods to select domains with an ADA risk for additional modification. The resultant engineered antibodies had a comparable affinity for TNFα, demonstrated similar biophysical properties, and exhibited significantly reduced ADA levels in cynomolgus macaque compared with the parental antibodies, with a corresponding improvement in the pharmacokinetic profile. Notably, plasma concentrations of the engineered antibodies were quantifiable through 504 hours (chimeric) and 840 hours (germlined/de-immunized), compared with only 336 hours (adalimumab) or 336-672 hours (golimumab). The results point to the significant value in the investment in these engineering strategies as an important guide for monoclonal antibody optimization that can contribute to improved clinical outcomes.

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Single-Domain Antibodies or Nanobodies: A Class of Next-Generation Antibodies

Cited by 89 publications

References 45 publications

Imaging the adenovirus infection cycle

Imaging the adenovirus infection cycle

Targeting RyR2 with a phosphorylation site–specific nanobody reverses dysfunction of failing cardiomyocytes in rats

Validation of a de‐immunization strategy for monoclonal antibodies using cynomolgus macaque as a surrogate for human

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