NeutrobodyPlex—monitoring SARS‐CoV‐2 neutralizing immune responses using nanobodies

Wagner, Teresa R.; Ostertag, E.; Kaiser, Philipp; Gramlich, Marius; Ruétalo, Natalia; Junker, Daniel; Haering, Julia; Traenkle, Bjoern; Becker, Matthias; Dulovic, Alex; Schweizer, H; Nueske, Stefan; Scholz, Armin M.; Zeck, Anne; Schenke‐Layland, Katja; Nelde, Annika; Strengert, Monika; Walz, Juliane S.; Zocher, Georg; Stehle, Thilo; Schindler, Michael; Schneiderhan‐Marra, Nicole; Rothbauer, Ulrich

doi:10.15252/embr.202052325

Cited by 48 publications

(50 citation statements)

References 77 publications

(156 reference statements)

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“…Wagner et al [ 42 ]. have identified eleven nanobodies isolated from an immunized Alpaca with high-affinity against RBD.…”

Section: Resultsmentioning

confidence: 99%

“…To investigate whether bipNb NM1267 can be a suitable nanobady for monitoring the presence and appearance of neutralizing antibodies in patients' serum, the NeutrobodyPlex method was applied. For this purpose, RBD, S1, S2, and Spike of SARSCoV-2 were incubated with NM1267 and also another well-specified nanobody as a control which targets the interface between RBD and ACE2 resulted in selection of NM1267 as a potent nanobody to detected neutralizing antibodies against RBD [ 42 ].…”

Section: Resultsmentioning

confidence: 99%

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Outlook of therapeutic and diagnostic competency of nanobodies against SARS-CoV-2: A systematic review

Aria

Mahmoodi

Ghaheh

et al. 2022

Analytical Biochemistry

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Purpose The newly emerged coronavirus (SARS-CoV-2) continues to infect humans, and no completely efficient treatment has yet been found. Antibody therapy is one way to control infection caused by COVID-19, but the use of classical antibodies has many disadvantages. Heavy chain antibodies (HCAbs) are single-domain antibodies derived from the Camelidae family. The variable part of these antibodies (Nanobodies or VHH) has interesting properties such as small size, identify criptic epitopes, stability in harsh conditions, good tissue permeability and cost-effective production causing nanobodies have become a good candidate in the treatment and diagnosis of viral infections. Methods Totally 157 records (up to November 10, 2021), were recognized to be reviewed in this study. 62 studies were removed after first step screening due to their deviation from inclusion criteria. The remaining 95 studies were reviewed in details. After removing articles that were not in the study area, 45 remaining studies met the inclusion criteria and were qualified to be included in the systematic review. Results In this systematic review, the application of nanobodies in the treatment and detection of COVID-19 infection was reviewed. The results of this study showed that extensive and sufficient studies have been performed in the field of production of nanobodies against SARS-CoV-2 virus and the obtained nanobodies have a great potential for use in patients infected with SARS-CoV-2 virus. Conclusion According to the obtained results, it was found that nanobodies can be used effectively in the treatment and diagnosis of SARS-CoV-2 virus.

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“…Wagner et al [ 42 ]. have identified eleven nanobodies isolated from an immunized Alpaca with high-affinity against RBD.…”

Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Outlook of therapeutic and diagnostic competency of nanobodies against SARS-CoV-2: A systematic review

Aria

Mahmoodi

Ghaheh

et al. 2022

Analytical Biochemistry

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“…Extensive research has demonstrated that nanobodies play a significant role in several inflammatory diseases. [44][45][46] In related experiments, nanobodies specific to TLR4 treatment significantly decreased mRNA and protein levels of proinflammatory cytokines in NR8383 cells. 47 Consistently with that research, our in vitro tests showed that compared with negative nanobody treatment, specific nanobody treatment significantly suppressed mRNA and protein expression of IL1β and IL6 in HCECs stimulated by A. fumigatus.…”

Section: Effects Of Nanobodies On Proliferation Of Hcecsmentioning

confidence: 91%

Preparation of a Nanobody Specific to Dectin 1 and Its Anti-inflammatory Effects on Fungal Keratitis

Liu¹,

Sui²,

Li³

et al. 2022

IJN

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To prepare a nanobody specific to dectin 1 and verify its specificity and anti-inflammatory effects on Aspergillus fumigatus keratitis. Methods: The nanobody was selected from a high-quality shark-antibody library constructed with phage-display technology. The nanobody was developed in the expression systems of Escherichia coli. Indirect ELISA was used to determine the specificity of the nanobody to recombinant dectin 1 protein. The potential of the nanobody to be recognized and expressed on the surfaces of cells and corneas was detected by immunofluorescence, and its anti-inflammatory effect on A. fumigatus keratitis was further verified. After infection with A. fumigatus, eyes of C57B L/6 mice were treated with nanobodies. Human corneal epithelial cells (HCECs) were pretreated with nanobodies and then incubated with A. fumigatus. Clinical scores and slit-lamp photography were used to assess disease response in mouse corneas. RT-PCR and ELISA were used to evaluate mRNA and protein expression of IL1β and IL6 in both mouse corneas and HCECs. Results: The nanobody was successfully expressed through microbial system and showed specific high-affinity binding to recombinant dectin 1. Furthermore, it exhibited specific binding to dectin 1 expressed on the surfaces of cells and recognized dectin 1 in mouse corneas. Importantly, it reduced clinical scores of A. fumigatus keratitis in mice compared with a PBS-treatment group. In addition, it decreased mRNA and protein expression of IL1β and IL6 in infected corneas and HCECs stimulated with A. fumigatus. Conclusion: These results suggest that this nanobody can bring about anti-inflammatory effects. This highlights the potential of these nanobodies as innovative therapeutic agents in A. fumigatus.

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“…CD4-specific Nbs were expressed and purified as previously published ( 71 , 75 ). Extracellular fragment of hCD4 comprising domains 1–4 of hCD4 and a C-terminal His6-tag was expressed in Expi293 cells according to the manufacturer’s protocol (TFS).…”

Section: Methodsmentioning

confidence: 99%

“…A peptic peptide list was generated in a preliminary LC-MS/MS experiment as described previously ( 75 ). For data-based search, no enzyme selectivity was applied; furthermore, identified peptides were manually evaluated to exclude peptides originated through cleavage after arginine, histidine, lysine, proline, and the residue after proline ( 79 ).…”

Section: Methodsmentioning

confidence: 99%

Single-Domain Antibodies for Targeting, Detection, and In Vivo Imaging of Human CD4+ Cells

et al. 2021

Self Cite

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The advancement of new immunotherapies necessitates appropriate probes to monitor the presence and distribution of distinct immune cell populations. Considering the key role of CD4+ cells in regulating immunological processes, we generated novel single-domain antibodies [nanobodies (Nbs)] that specifically recognize human CD4. After in-depth analysis of their binding properties, recognized epitopes, and effects on T-cell proliferation, activation, and cytokine release, we selected CD4-specific Nbs that did not interfere with crucial T-cell processes in vitro and converted them into immune tracers for noninvasive molecular imaging. By optical imaging, we demonstrated the ability of a high-affinity CD4-Nb to specifically visualize CD4+ cells in vivo using a xenograft model. Furthermore, quantitative high-resolution immune positron emission tomography (immunoPET)/MR of a human CD4 knock-in mouse model showed rapid accumulation of 64Cu-radiolabeled CD4-Nb1 in CD4+ T cell-rich tissues. We propose that the CD4-Nbs presented here could serve as versatile probes for stratifying patients and monitoring individual immune responses during personalized immunotherapy in both cancer and inflammatory diseases.

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NeutrobodyPlex—monitoring SARS‐CoV‐2 neutralizing immune responses using nanobodies

Cited by 48 publications

References 77 publications

Outlook of therapeutic and diagnostic competency of nanobodies against SARS-CoV-2: A systematic review

Outlook of therapeutic and diagnostic competency of nanobodies against SARS-CoV-2: A systematic review

Preparation of a Nanobody Specific to Dectin 1 and Its Anti-inflammatory Effects on Fungal Keratitis

Single-Domain Antibodies for Targeting, Detection, and In Vivo Imaging of Human CD4+ Cells

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