Diverse human VH antibody fragments with bio-therapeutic properties from the Crescendo Mouse

Teng, Yumin; Young, Joyce L.; Edwards, Bryan; Hayes, Philip J.; Thompson, Lorraine; Johnston, C L; Edwards, C. R. W.; Sanders, Yun; Writer, Michele; Pinto, Débora; Zhang, Yanjing; Roode, Mila; Chovanec, Peter; Matheson, Louise S.; Corcoran, Anne E.; Fernández, Almudena; Montoliu, Lluı́s; Rossi, Brunella; Tosato, Valentina; Gjuračić, Krešimir; Nikitin, Dmitri; Bruschi, Carlo V.; McGuinness, Brian; Sandal, Thomas; Romanos, Michael A.

doi:10.1016/j.nbt.2019.10.003

Cited by 25 publications

(26 citation statements)

References 49 publications

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“…A key contribution of our study is the creation of Nb-producing mice. Previous work explored transgenic expression of 1-2 llama VHHs or human VHs in Igh -/- Igκ -/- Igλ -/- mice (Drabek et al, 2016; Janssens et al, 2006; Teng et al, 2020). In our model, the 30 VHHs replace the entire VH domain leading to physiological recombination and selection during B cell development.…”

Section: Discussionmentioning

confidence: 99%

Multimeric nanobodies from camelid engineered mice and llamas potently neutralize SARS-CoV-2 variants

Xu²,

Jung

et al. 2021

Preprint

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Since the start of the coronavirus disease-2019 (COVID-19) pandemic, severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) has caused more than 2 million deaths worldwide. Many vaccines have been deployed to date, but the continual evolution of the viral receptor-binding domain (RBD) has recently challenged their efficacy. In particular, SARS-CoV-2 variants originating in the U.K. (B.1.1.7), South Africa (B.1.351) and New York (B.1.526) have reduced neutralization activity from convalescent sera and compromised the efficacy of some antibody cocktails that received emergency use authorization. Whereas vaccines can be updated periodically to account for emerging variants, complementary strategies are urgently needed to avert viral escape. One potential alternative is the use of camelid VHHs (also known as nanobodies), which due to their small size can recognize protein domains that are often inaccessible to conventional antibodies. Here, we isolate anti-RBD nanobodies from llamas and nanomice we engineered to produce VHHs cloned from alpacas, dromedaries and camels. Through binding assays and cryo-electron microscopy, we identified two sets of highly neutralizing nanobodies. The first group expresses VHHs that circumvent RBD antigenic drift by recognizing a region outside the ACE2-binding site that is conserved in coronaviruses but is not typically targeted by monoclonal antibodies. The second group is almost exclusively focused to the RBD-ACE2 interface and fails to neutralize pseudoviruses carrying the E484K or N501Y substitutions. Notably however, they do neutralize the RBD variants when expressed as homotrimers, rivaling the most potent antibodies produced to date against SARS-CoV-2. These findings demonstrate that multivalent nanobodies overcome SARS-CoV-2 variant mutations through two separate mechanisms: enhanced avidity for the ACE2 binding domain, and recognition of conserved epitopes largely inaccessible to human antibodies. Therefore, while new SARS-CoV-2 mutants will continue to emerge, nanobodies represent promising tools to prevent COVID-19 mortality when vaccines are compromised.

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Section: Discussionmentioning

confidence: 99%

Multimeric nanobodies from camelid engineered mice and llamas potently neutralize SARS-CoV-2 variants

Xu²,

Jung

et al. 2021

Preprint

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“…However, the limitations of an ADC may be the incomplete penetration into cancer cells and solid tumor, due to the large size of the intact Ab (Xenaki et al, 2017). More recently, VH Abs from many species have emerged as a smaller targeting molecule (Kontermann et al, 2009;Hairul Bahara et al, 2016;Keyaerts et al 2016;Teng et al, 2020). The idea behind these studies was based upon the advantages of VH of its small size resulting in a good penetration into the desired cells (Arbabi Ghahroudi et al, 1997;Harmsen & De Haard, 2007) .…”

Section: Discussionmentioning

confidence: 99%

Development of a human antibody fragment directed against the alpha folate receptor as a promising molecule for targeted application

et al. 2021

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Alpha folate receptor (FRa) is currently under investigation as a target for the treatment of patients with non-small-cell lung cancer (NSCLC), since it is highly expressed in tumor cells but is largely absent in normal tissue. In this study, a novel human variable domain of a heavy-chain (VH) antibody fragment specific to FRa was enriched and selected by phage bio-planning. The positive phage clone (3A102 VH) specifically bound to FRa and also cross-reacted with FRb, as tested by ELISA. Clone 3A102 VH was then successfully expressed as a soluble protein in an E. coli shuffle strain. The obtained soluble 3A102 VH demonstrated a high affinity for FRa with affinity constants (K aff ) values around 7.77 ± 0.25 Â 10 7 M À1 , with specific binding against both FRa expressing NSCLC cells and NSCLC patient-derived primary cancer cells, as tested by cell ELISA. In addition, soluble 3A102 VH showed the potential desired property of a targeting molecule by being internalized into FRa-expressing cells, as observed by confocal microscopy. This study inspires the use of phage display to develop human VH antibody (Ab) fragments that might be well suited for drug targeted therapy of NSCLC and other FRapositive cancer cells.

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“…Apart from immunising a camelid, several groups have invested in generating a transgenic mouse producing HCAbs. Obviously, these transgenic mice are a good substitute for camelids in cases where the Ag is difficult to obtain, and indeed, good single‐domain antibodies (sdAbs) have been retrieved following immunising such mice [35,36].…”

Section: Generation Of Nanobody Librariesmentioning

confidence: 99%

A guide to: generation and design of nanobodies

2020

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Immune or naïve nanobody (Nb) repertoires are cloned from B lymphocytes of camelids. Alternatively, synthetic Nb libraries are generated in vitro by randomising codons forming antigen‐binding loops of a Nb scaffold. These libraries serve to retrieve target‐specific Nbs by phage display pannings, yeast surface display or other selection methods. After identification of the lead Nb and possible engineering efforts, multiple applications have been explored in research, diagnosis or therapy.

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Diverse human VH antibody fragments with bio-therapeutic properties from the Crescendo Mouse

Cited by 25 publications

References 49 publications

Multimeric nanobodies from camelid engineered mice and llamas potently neutralize SARS-CoV-2 variants

Multimeric nanobodies from camelid engineered mice and llamas potently neutralize SARS-CoV-2 variants

Development of a human antibody fragment directed against the alpha folate receptor as a promising molecule for targeted application

A guide to: generation and design of nanobodies

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