The DOI matches its Crossref record; 96 of the 96 references it cites were found in a registry or an index.
Properties, production, and applications of camelid single-domain antibody fragments
Open accessAbstract
Camelids produce functional antibodies devoid of light chains of which the single N-terminal domain is fully capable of antigen binding. These single-domain antibody fragments (VHHs or Nanobodies®) have several advantages for biotechnological applications. They are well expressed in microorganisms and have a high stability and solubility. Furthermore, they are well suited for construction of larger molecules and selection systems such as phage, yeast, or ribosome display. This minireview offers an overview of (1) their properties as compared to conventional antibodies, (2) their production in microorganisms, with a focus on yeasts, and (3) their therapeutic applications.
Overview
The claims and statistics of this paper haven’t been checked yet.
Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.
Integrity signals
Registry
Cites
Statistics
Verification
Claims the paper makes
The claims and statistics of this paper haven’t been checked yet.
Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.
Claims other papers make about this one
Each is checked against this paper’s own text: does it say what the citing paper says it does?
The cited source says what the claim attributes to it.
In the citing paper
“Given their high-affinity, solubility and stability also in absence of the partner VL domain, the camelid VH domains (VHH) [4] and the shark VH domains called V-NAR (New Antigen Receptor) [5], are currently used as templates to generate highly efficient and stable new antibody fragments collectively termed nanobodies [6,7,8].”
From Evolution of Escherichia coli Expression System in Producing Antibody Recombinant Fragments (2020) · cites this paper as [4]
Self-check
Percentages, sums and reported differences re-computed from the paper’s own numbers.
The self-check needs the paper’s full text.
Citations
Cites
Naturally occurring antibodies devoid of light chains
Hamers-Casterman, C., Atarhouch, T., Muyldermans, S., Robinson, G., Hammers, C., Songa, E. B., Bendahman, N., & Hammers, R. (1993) · Nature · doi:10.1038/363446a0 · Cited by 3,241 · cited 4×
“Sequence analysis (Hamers-Casterman et al. 1993; Muyldermans et al. 1994; Vu et al. 1997; Harmsen et al. 2000) and elucidation of the crystal structure (Desmyter et al.”
BackgroundInfluentialRapid generation of functional human IgG antibodies derived from Fab-on-phage display libraries
Jostock, T., Vanhove, M., Brepoels, E., van Gool, R., Daukandt, M., Wehnert, A., van Hegelsom, R., Dransfield, D., Sexton, D., Devlin, M., et al. (2004) · Journal of Immunological Methods · doi:10.1016/j.jim.2004.03.014 · Cited by 2,572
Engineered antibody fragments and the rise of single domains
Holliger, P., & Hudson, P. J. (2005) · Nature Biotechnology · doi:10.1038/nbt1142 · Cited by 1,928 · cited 1×
“As compared to human VH domains, the VHH FRs show a high sequence homology of more than 80%, and their 3D structures can be superimposed (Muyldermans et al. 2001; Holliger and Hudson 2005).”
BackgroundWard, E. S., Güssow, D., Griffiths, A. D., Jones, P., & Winter, G. P. (1989) · Nature · doi:10.1038/341544a0 · Cited by 1,010 · cited 2×
“Although isolated antibody heavy (Utsumi and Karush 1964) and light chains (Yoo et al. 1967) can retain antigenbinding specificity, their affinity and solubility is often reduced (Ward et al. 1989).”
BackgroundGreenberg, A. S., Avila, D. W., Hughes, M. K., Hughes, A. L., McKinney, E. C., & Flajnik, M. F. (1995) · Nature · doi:10.1038/374168a0 · Cited by 783 · cited 2×
“Although single-domain antibodies later were also identified in particular cartilaginous fish (Greenberg et al. 1995), most research on the biotechnological application of singledomain antibodies was done using camelids because of Appl Microbiol Biotechnol (2007) 77:13–22”
BackgroundSelection and identification of single domain antibody fragments from camel heavy‐chain antibodies
Ghahroudi, M. A., Desmyter, A., Wyns, L., HAMERS, R., & Muyldermans, S. (1997) · FEBS Letters · doi:10.1016/s0014-5793(97)01062-4 · Cited by 775 · cited 1×
“Methods to isolate antigen-specific VHHs from immune ( Arbabi-Ghahroudi et al. 1997; Van der Linden et al. 2000a), nonimmune (Tanha et al. 2002; Yau et al. 2003; Verheesen et al. 2006), or semisynthetic (Goldman et al. 2006) libraries using phage, yeast, or ribosome display are now well established.”
BackgroundTargeting and tracing antigens in live cells with fluorescent nanobodies
Rothbauer, U., Zolghadr, K., Tillib, S. V., Nowak, D., Schermelleh, L., Gahl, A., Backmann, N., Conrath, K., Muyldermans, S., Cardoso, M. C., et al. (2006) · Nature Methods · doi:10.1038/nmeth953 · Cited by 715 · cited 1×
“Because of their stability, VHHs are especially suited for intrabody production because this requires expression in the reducing environment of the cytoplasm (Gueorguieva et al. 2006; Rothbauer et al. 2006).”
BackgroundMolecular basis for the preferential cleft recognition by dromedary heavy-chain antibodies
Genst, E. D., Silence, K., Decanniere, K., Conrath, K., Loris, R., Kinne, J., Muyldermans, S., & Wyns, L. (2006) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.0505379103 · Cited by 681 · cited 4×
“Occasionally, VHHs with affinity constants (KD) as low as 100 pM are isolated (Saerens et al. 2004; De Genst et al. 2006; Harmsen et al. 2006), which equals the affinity ceiling proposed for natural antibodies (Sundberg and Mariuzza 2002).”
BackgroundInfluentialSingle‐domain antibody fragments with high conformational stability
Dumoulin, M., Conrath, K., Meirhaeghe, A. V., Meersman, F., Heremans, K., Frenken, L. G. J., Muyldermans, S., Wyns, L., & Matagne, A. (2002) · Protein Science · doi:10.1110/ps.34602 · Cited by 641 · cited 4×
“VHHs are especially suited for oral immunotherapy because of their resistance against extremes of pH and the capacity to bind to the target at high concentrations of chaotropic agents (Dumoulin et al. 2002, 2003).”
BackgroundInfluentialStructure-Immunogenicity Relationships of Therapeutic Proteins
Hermeling, S., Crommelin, D. J. A., Schellekens, H., & Jiskoot, W. (2004) · Pharmaceutical Research · doi:10.1023/b:pham.0000029275.41323.a6 · Cited by 617 · cited 1×
“This could rely on their high sequence homology to conventional VH domains and on their high stability because aggregation of proteins is known to increase immunogenicity (Hermeling et al. 2004).”
Background
Flagged first, then by citations
Research works printed but not linked: 1
- Estell D (2006) Adapting industry practices for the rapid, large-scale manufacture of pharmaceutical proteins. Bridge 36:39–44
Cited by
Sorted by citations
Items 301–400 of 809
van Lith, S. A. M., van den Brand, D., Wallbrecher, R., van Duijnhoven, S. M. J., Brock, R. E., & Leenders, W. P. J. (2017) · ChemBioChem · doi:10.1002/cbic.201700444 · Cited by 28
Shahapurmath, S., Sharannavar, B. R., & Koli, R. (2025) · Medicine in Drug Discovery · doi:10.1016/j.medidd.2025.100218 · Cited by 29
Expansion of the aptamer library from a “natural soup” to an “unnatural soup”
Uzawa, T., Tada, S., Wang, W., & Ito, Y. (2013) · Chemical Communications · doi:10.1039/c2cc36348h · Cited by 27
Hambach, J., Fumey, W., Stähler, T., Gebhardt, A. J., Adam, G., Weisel, K. C., Koch‐Nolte, F., & Bannas, P. (2022) · Frontiers in Immunology · doi:10.3389/fimmu.2022.838406 · Cited by 31
Shriver‐Lake, L. C., Zabetakis, D., Goldman, E. R., & Anderson, G. P. (2017) · Toxicon · doi:10.1016/j.toxicon.2017.06.002 · Cited by 27
Oligoclonal selection of nanobodies targeting vascular endothelial growth factor
Ahadi, M., Ghasemian, H., Behdani, M., & Kazemi‐Lomedasht, F. (2018) · Journal of Immunotoxicology · doi:10.1080/1547691x.2018.1526234 · Cited by 27
“These findings, and the fact that single domain entity nanobodies make them good options for use in antibody mixtures (Harmsen and De Haard 2007), have led to the performance of several studies using mixtures of nanobodies (oligoclonal mode) to target distinct epitopes on tumor cells.”
BackgroundVerma, A. K., Khan, E., Bhagwat, S. R., & Kumar, A. (2019) · Molecular Neurobiology · doi:10.1007/s12035-019-01724-4 · Cited by 28
Considerations on the stability of IgG antibody in clinical specimens
Yen, L., Henao-Díaz, A., Zimmerman, J. J., & Gimenez-Lirola, L. G. (2024) · Journal of Veterinary Diagnostic Investigation · doi:10.1177/10406387241296848 · Cited by 28
“49,130 In cows, antibodies also differ from the general mammalian IgG structure; the extra-long third complementarity-determining regions in the heavy chain were originally observed in IgM114 and later determined to be a trend existing across isotypes.”
Can Intrabodies Serve as Neuroprotective Therapies for Parkinson's Disease? Beginning Thoughts
Bhatt, M. A., Messer, A. C., & Kordower, J. H. (2013) · Journal of Parkinson s Disease · doi:10.3233/jpd-130252 · Cited by 26
“Camelids synthesize an especially small and stable VH nanobody fragment, with several of these molecules in current clinical trials [36–39].”
Günaydın, G., Álvarez, B., Lin, Y. Y., Hammarström, L. L. G., & Marcotte, H. (2014) · PLoS ONE · doi:10.1371/journal.pone.0096409 · Cited by 26
“In addition to its antigen binding capacity similar to that of complete antibodies, their acid resistance property contributes to long term stability in the harsh gastrointestinal environment [17].”
BackgroundGodar, M., Morello, V., Sadi, A., Hultberg, A., Jonge, N. D., Basilico, C., Hanssens, V., Saunders, M. J., Lambrecht, B. N., Khattabi, M. E., et al. (2016) · Scientific Reports · doi:10.1038/srep31621 · Cited by 28
Camelid VHH affinity ligands enable separation of closely related biopharmaceuticals
Pabst, T. M., Wendeler, M., Wang, X., Bezemer, S., Hermans, P., & Hunter, A. K. (2016) · Biotechnology Journal · doi:10.1002/biot.201600357 · Cited by 26
“These affinity ligands consist of single monomeric variable domains of camelid heavy-chain only antibodies (VHHs) that can efficiently be expressed at large scale in baker’s yeast free of Research Article Camelid VHH affinity ligands enable separation of closely related biopharmaceuticals Timothy…”
BackgroundRaeisi, H., Azimirad, M., Nabavi-Rad, A., Aghdaei, H. A., Yadegar, A., & Zali, M. R. (2022) · Frontiers in Immunology · doi:10.3389/fimmu.2022.972930 · Cited by 28
Harish, P., Malerba, A., Dickson, G., & Bachtarzi, H. (2015) · Human Gene Therapy · doi:10.1089/hum.2015.014 · Cited by 26
“50,51 Nanobodies were tested as therapeutics for OPMD both in cellular models and in Drosophila .”
Medecigo, M., Manoutcharian, K., Vasilevko, V., Govezensky, T., Munguía, M. E., Becerril, B., Luz-Madrigal, A., Vaca, L. A., Cribbs, D. H., & Gevorkian, G. (2010) · Journal of Neuroimmunology · doi:10.1016/j.jneuroim.2010.03.023 · Cited by 26
“…protein transgenic (APP/Tg) mouse models, which develop AD-like amyloid plaque pathology (Schenk et al., 1999; Bard et al., 2000; Janus et al., 2000; Hartman et al., 2005; Morgan, 2006; Brody and Holtzman, 2008), as well as in canine and primate models of amyloidosis (Lemere et al., 2004, Head et…”
Specific immuno capturing of the Staphylococcal superantigen toxic‐shock syndrome toxin‐1 in plasma
Adams, H., Brummelhuis, W. J., Maassen, B., van Egmond, N., Khattabi, M. E., Detmers, F. J. M., Hermans, P., Braam, B. G., Stam, J. C., & Verrips, T. C. (2009) · Biotechnology and Bioengineering · doi:10.1002/bit.22365 · Cited by 26
Wilken, L., & McPherson, A. (2017) · International Reviews of Immunology · doi:10.1080/08830185.2017.1397657 · Cited by 26
IgGM: A Generative Model for Functional Antibody and Nanobody DesignPreprint
Wang, R., Wu, F., Gao, X., Wu, J., Zhao, P., & Yao, J. (2024) · bioRxiv (Cold Spring Harbor Laboratory) · doi:10.1101/2024.09.19.613838 · Cited by 27
“Nanobody sources are a type of single-domain antibody known as VHH fragments (Harmsen & De Haard, 2007).”
Application of nanotechnology in CAR-T-cell immunotherapy
Zeng, Q., Liu, Z., Niu, T., He, C., Qu, Y., & Qian, Z. (2022) · Chinese Chemical Letters · doi:10.1016/j.cclet.2022.107747 · Cited by 25
Targeting the Brain with Single-Domain Antibodies: Greater Potential Than Stated So Far?
Tsitokana, M. E., Lafon, P., Prézeau, L., Pin, J., & Rondard, P. (2023) · International Journal of Molecular Sciences · doi:10.3390/ijms24032632 · Cited by 25
Development of Oligoclonal Nanobodies for Targeting the Tumor-Associated Glycoprotein 72 Antigen
Sharifzadeh, Z., Rahbarizadeh, F., Shokrgozar, M. A., Ahmadvand, D., Mahboudi, F., Jamnani, F. R., & Aghaee‐Bakhtiari, S. H. (2012) · Molecular Biotechnology · doi:10.1007/s12033-012-9601-0 · Cited by 25
“These unusual immunoglobulins lack the light chains and CH1 domain; thereby their antigen-binding site is reduced to a single variable domain [16].”
BackgroundInfluentialTrilling, A. K., de Ronde, H., Noteboom, L., van Houwelingen, A. M. M. L., Roelse, M., Srivastava, S. K., Haasnoot, W., Jongsma, M. A., Kolk, A. H. J., Zuilhof, H. T., et al. (2011) · PLoS ONE · doi:10.1371/journal.pone.0026754 · Cited by 25
“Llama antibody fragments (VHHs) are particularly suited for these applications due to their compact size (15 kDa) [9] and remarkable physicochemical stability [10–12].”
BackgroundGeneration of single-domain antibody multimers with three different self-associating peptides
Wang, L., Liu, X., Zhu, X., Wang, L., Wang, W., Liu, C., Cui, H., Sun, M., & Gao, B. (2013) · Protein Engineering Design and Selection · doi:10.1093/protein/gzt011 · Cited by 25
Demeestere, D., Dejonckheere, E., Steeland, S., Hulpiau, P., Haustraete, J., Devoogdt, N., Wichert, R., Becker‐Pauly, C., Wonterghem, E. V., Dewaele, S., et al. (2016) · Molecular Therapy · doi:10.1038/mt.2016.2 · Cited by 25
In planta expression of nanobody-based designer chicken antibodies targeting Campylobacter
Vanmarsenille, C., Elseviers, J., Yvanoff, C., Hassanzadeh‐Ghassabeh, G., García, G. G.-J., Martens, E., Depicker, A., Martel, A., Haesebrouck, F., Pasmans, F., et al. (2018) · PLoS ONE · doi:10.1371/journal.pone.0204222 · Cited by 25
Zhu, L., Huang, B., Wang, X., Ni, F., Ao, M., Wang, R., Zheng, B., Chen, C., Xue, J., Zhu, L., et al. (2024) · Nature Communications · doi:10.1038/s41467-024-51414-6 · Cited by 25
“Derived from the Camelidae family, nanobodies constitute the smallest naturally functional antibody fragment available to date 10 – 13 , making it possible to create multi-targeted antibodies.”
Proteomic strategies and challenges in tumor metastasis research
Hudler, P., Goršič, M., & Komel, R. (2010) · Clinical & Experimental Metastasis · doi:10.1007/s10585-010-9339-7 · Cited by 24
“In our and some other laboratories variable domains of single-chain antibodies, naturally occurring in camelidae (named also nano-bodies), were generated for immobilization on microarray surface [73, 74].”
BackgroundMaggi, M., & Scotti, C. (2017) · Protein Expression and Purification · doi:10.1016/j.pep.2017.02.007 · Cited by 24
Evaluation the potential of recombinant anti-CD3 nanobody on immunomodulatory function
Moradi‐Kalbolandi, S., Sharifi-K, A., Darvishi, B., Majidzadeh‐A, K., Jalili, N., Sadeghi, S., Mosayebzadeh, M., Sanati, H., Salehi, M., & Farahmand, L. (2019) · Molecular Immunology · doi:10.1016/j.molimm.2019.12.017 · Cited by 25
Truong, T. T. T., Huynh, V. Q., Vo, N. T., & Nguyen, H. D. (2022) · Journal of Genetic Engineering and Biotechnology · doi:10.1186/s43141-022-00439-9 · Cited by 24
“Multiple methods have been developed and utilized for screening the best Ab [5, 24, 28, 39] and Nb candidates [10, 12, 30] for therapeutic properties.”
MethodMIP-based immunoassays: A critical review
Nardo, F. D., Anfossi, L., & Baggiani, C. (2023) · Analytica Chimica Acta · doi:10.1016/j.aca.2023.341547 · Cited by 24
Head‐to‐tail fusions of camelid antibodies can be expressed in planta and bind in rumen fluid
Winichayakul, S., Pernthaner, A., Scott, R., Vlaming, R., & Roberts, N. (2008) · Biotechnology and Applied Biochemistry · doi:10.1042/ba20080076 · Cited by 24
High-content 3D multicolor super-resolution localization microscopy
Pereira, P. M., Almada, P., & Henriques, R. (2015) · Methods in cell biology · doi:10.1016/bs.mcb.2014.10.004 · Cited by 25
Vanmarsenille, C., Diaz-del-Olmo, I., Elseviers, J., Ghassabeh, G. H., Moonens, K., Vertommen, D., Martel, A., Haesebrouck, F., Pasmans, F., Hernálsteens, J.-P., et al. (2017) · Veterinary Research · doi:10.1186/s13567-017-0491-9 · Cited by 24
“Nanobodies interact with their antigen with high affinity and specificity, they are highly stable and soluble and can be expressed in microbial expression systems [32, 33].”
BackgroundZafra, O., Fraile, S., Gutiérrez, C., Haro, A., Páez-Espino, D., Jiménez, J. I., & de Lorenzo, V. (2011) · Environmental Microbiology · doi:10.1111/j.1462-2920.2010.02401.x · Cited by 24
Pereira, S. D. S., Moreira-Dill, L. S., Morais, M. S. D. S., Prado, N. D. R., Barros, M. L., Koishi, A. C., Mazzarotto, G. A. C. A., Gonçalves, G. M., Zuliani, J. P., Calderón, L. A., et al. (2014) · PLoS ONE · doi:10.1371/journal.pone.0108067 · Cited by 23
“In addition to conventional antibodies, camelids produce functional immunoglobulins composed only of heavy chains in which the antigen binding site is formed only by the single Nterminal variable domain, referred to as VHH [5], [6], [7].”
BackgroundChen, G., Li, Z., Theile, C. S., Bardhan, N. M., Kumar, P. V., Duarte, J. N., Maruyama, T., Rashidfarrokh, A., Belcher, A. M., & Ploegh, H. L. (2015) · Chemistry - A European Journal · doi:10.1002/chem.201503057 · Cited by 23
Suzuki, T., Mochizuki, Y., Kimura, S., Ogawa, Y., Hagihara, Y., & Nemoto, N. (2018) · Biochemical and Biophysical Research Communications · doi:10.1016/j.bbrc.2018.07.158 · Cited by 23
Criscitiello, M. F., Kraev, I., & Lange, S. (2019) · Molecular Immunology · doi:10.1016/j.molimm.2019.10.017 · Cited by 23
Radiolabeled nanobodies for tumor targeting: From bioengineering to imaging and therapy
Piramoon, M., Khodadust, F., & Hosseinimehr, S. J. (2021) · Biochimica et Biophysica Acta (BBA) - Reviews on Cancer · doi:10.1016/j.bbcan.2021.188529 · Cited by 23
Development of potent and effective synthetic SARS-CoV-2 neutralizing nanobodies
Stefan, M. A., Light, Y. K., Schwedler, J. L., McIlroy, P. R., Courtney, C. M., Saada, E. A., Thatcher, C. E., Phillips, A. M., Bourguet, F. A., Mageeney, C. M., et al. (2021) · mAbs · doi:10.1080/19420862.2021.1958663 · Cited by 25
Production of Camel-Like Antibodies in Plants
Buck, S. D., Virdi, V., Meyer, T. D., Wilde, K. D., Piron, R., Nolf, J., Lerberge, E. V., Paepe, A. D., & Depicker, A. (2012) · Methods in molecular biology · doi:10.1007/978-1-61779-968-6_19 · Cited by 23
Harmsen, M. M., & Fijten, H. (2012) · Journal of Immunoassay and Immunochemistry · doi:10.1080/15321819.2011.634473 · Cited by 23
“[ 4 , 5 , 9 ] These comprise ease of genetic manipulation, amenability to phage display selection, efficient production of multivalent formats, inherent physicochemical and proteolytic stability, high microbial production levels, structural aptness for recognition of cryptic antigenic sites such as enzyme active sites…”
BackgroundSelf-citationTraxlmayr, M. W., Wozniak‐Knopp, G., Antes, B., Stadlmayr, G., Rüker, F., & Obinger, C. (2011) · Journal of Biotechnology · doi:10.1016/j.jbiotec.2011.06.042 · Cited by 22
Bioengineering of Bacteria To Assemble Custom-Made Polyester Affinity Resins
Hay, I. D., Du, J., Burr, N. S., & Rehm, B. H. A. (2014) · Applied and Environmental Microbiology · doi:10.1128/aem.02595-14 · Cited by 22
“VHH domains with specific affinities can be derived from immune, nonimmune, or semisynthetic libraries (13).”
BackgroundToffoli, E. C., Sheikhi, A., Lameris, R., King, L. A., van Vliet, A. A., Walcheck, B. K., Verheul, H. M. W., Spanholtz, J., Tuynman, J. B., de Gruijl, T. D., et al. (2021) · Cancers · doi:10.3390/cancers13215446 · Cited by 23
Characterization of camel nanobodies specific for superfolder GFP fusion proteins
Twair, A., Al-Okla, S., Zarkawi, M., & Abbady, A. Q. (2014) · Molecular Biology Reports · doi:10.1007/s11033-014-3575-x · Cited by 22
“Conversely, the libraries generated from the immunized camels show fully functional diversity due to the single domain entity of nanobodies [34].”
BackgroundGünaydın, G., Yu, S., Gräslund, T., Hammarström, L. L. G., & Marcotte, H. (2016) · Scientific Reports · doi:10.1038/srep30171 · Cited by 22
Velásquez, A. C., Nomura, K., Cooper, M. D., Herrin, B. R., & He, S. Y. (2017) · Plant Methods · doi:10.1186/s13007-017-0180-8 · Cited by 22
“Nanobodies®, the recombinant variable binding domain of heavy-chain only antibodies (VHH) from Camelids [43] offer another alternative to target plant proteins.”
BackgroundAmino-Acid-Catalyzed Direct Aldol Bioconjugation
Howard, T. S., Cohen, R. D., Nwajiobi, O., Muneeswaran, Z. P., Sim, Y. E., Lahankar, N., Yeh, J. T. ‐., & Raj, M. (2018) · Organic Letters · doi:10.1021/acs.orglett.8b02265 · Cited by 22
Wöll, S., Bachran, C., Schiller, S., Schröder, M., Conrad, L., Swee, L. K., & Scherließ, R. (2018) · European Journal of Pharmaceutics and Biopharmaceutics · doi:10.1016/j.ejpb.2018.09.017 · Cited by 22
Tillib, S. V., Privezentseva, M. E., Ivanova, T. I., Vasilev, L. F., Efimov, G. A., Gursky, Y. G., Georgiev, G. P., Goldman, I. L., & Садчикова, Е. Р. (2014) · Journal of Chromatography B · doi:10.1016/j.jchromb.2013.12.034 · Cited by 22
Darvish, M., Behdani, M., Shokrgozar, M. A., Bagheri, K. P., & Shahbazzadeh, D. (2015) · Molecular Immunology · doi:10.1016/j.molimm.2015.10.002 · Cited by 21
Li, S., Jiang, K., Wang, T., Zhang, W., Shi, M., Chen, B., & Hua, Z. (2020) · Biotechnology Letters · doi:10.1007/s10529-020-02823-2 · Cited by 21
Camels’ biological fluids contained nanobodies: promising avenue in cancer therapy
Al‐Numair, N. S., Theyab, A., Alzahrani, F. A., Shams, A. M., Alanazi, I. O., Oyouni, A. A. A., Al‐Amer, O. M., Mavromatis, C., Saadeldin, I. M., Abdali, W. A., et al. (2022) · Cancer Cell International · doi:10.1186/s12935-022-02696-7 · Cited by 24
“Moreover, camel milk expressed distinctive immunoglobulins characterized as VHH antibodies or nanobodies [20].”
BackgroundNguyen, T. D., Bordeau, B. M., & Balthasar, J. P. (2023) · Molecular Cancer Therapeutics · doi:10.1158/1535-7163.mct-22-0804 · Cited by 21
Niazi, S. K., Magoola, M., & Mariam, Z. (2024) · Pharmaceuticals · doi:10.3390/ph17060741 · Cited by 26
“Single-domain antibodies can identify and bind selectively to A β oligomers in vitro, inhibit fibril formation, and prevent A β -induced neurotoxicity [87]. dAbs targeting tau protein aggregates have been investigated for their potential to halt their formation and spread, a critical aspect of AD.”
BackgroundSelection and Characterization of Specific Nanobody Against Human Immunoglobulin G
Kazemi‐Lomedasht, F., Behdani, M., Rahimpour, A., Habibi‐Anbouhi, M., Bagheri, K. P., & Shahbazzadeh, D. (2015) · Monoclonal Antibodies in Immunodiagnosis and Immunotherapy · doi:10.1089/mab.2014.0086 · Cited by 21
“Moreover, the elongated size of CDR3 loop (17 residue in VHH and 9–12 residue in VH) causes the enhanced solubility and stability of nanobodies in aqueous solutions.((4)) In addition, the nanobody gene can easily be cloned and expressed in bacteria and yeasts in high yield.”
BackgroundAffinity proteins and their generation
Ståhl, S., Kronqvist, N., Jonsson, A., & Löfblom, J. (2012) · Journal of Chemical Technology & Biotechnology · doi:10.1002/jctb.3929 · Cited by 22
Bagheri, M., Babaei, E., Shahbazzadeh, D., Habibi‐Anbouhi, M., Alirahimi, E., Kazemi‐Lomedasht, F., & Behdani, M. (2016) · Toxin Reviews · doi:10.1080/15569543.2016.1244552 · Cited by 21
Nanobodies—Useful Tools for Allergy Treatment?
Flicker, S., Zettl, I., & Tillib, S. V. (2020) · Frontiers in Immunology · doi:10.3389/fimmu.2020.576255 · Cited by 21
Production of biologically active scFv and VHH antibody fragments in Bifidobacterium longum
Shkoporov, A. N., Khokhlova, E. V., Savochkin, K. A., Кафарская, Л. И., & Efimov, B. A. (2015) · FEMS Microbiology Letters · doi:10.1093/femsle/fnv083 · Cited by 21
Dangerous scorpion fauna of Mali
Goyffon, M., Dabo, A. J., Coulibaly, S., Togo, G., & Chippaux, J. (2012) · The Journal of venomous animals and toxins including tropical diseases · doi:10.1590/s1678-91992012000400003 · Cited by 20
“Recombinant antibodies are also very interesting (13).”
Engineering of single-domain antibodies for next-generation snakebite antivenoms
Fernandes, C. F. C., Pereira, S. D. S., Luiz, M. B., Silva, N. K. R. L., Silva, M. C. S., Marinho, A. C. M., Fonseca, M. H. G., Furtado, G. P., Trevizani, R., Nicolete, R., et al. (2021) · International Journal of Biological Macromolecules · doi:10.1016/j.ijbiomac.2021.06.043 · Cited by 19
Arai, T., Wada, M., Nishiguchi, H., Takimura, Y., & Ishii, J. (2023) · Microbial Cell Factories · doi:10.1186/s12934-023-02109-y · Cited by 21
“Other important characteristics of nanobodies are their low molecular weight, high resistance to proteolysis and thermal denaturation, high solubility, and ease of production by microorganisms [15, 16].”
BackgroundJangra, P., & Singh, A. (2010) · Asian Pacific Journal of Tropical Medicine · doi:10.1016/s1995-7645(10)60020-x · Cited by 20
Stasevich, T. J., Sato, Y., Nozaki, N., & Kimurâ, H. (2014) · Methods · doi:10.1016/j.ymeth.2014.08.002 · Cited by 19
Thompson, M. K., Fridy, P. C., Keegan, S., Chait, B. T., Fenyö, D., & Rout, M. P. (2016) · Journal of Immunological Methods · doi:10.1016/j.jim.2016.01.006 · Cited by 21
Gao, X., Hu, X., Tong, L., Liu, D., Chang, X., Wang, H., Dang, R., Wang, X., Xiao, S., Du, E., et al. (2016) · BMC Veterinary Research · doi:10.1186/s12917-016-0664-1 · Cited by 20
“The presence of variable single domains that seem to originated from VH have been repeatedly identified together with canonical VHHs in immune libraries [35, 36].”
BackgroundChow, K. M., Whiteheart, S. W., Smiley, J. R., Sharma, S., Boaz, K., Coleman, M. J., Maynard, A., Hersh, L. B., & Kooi, C. W. V. (2019) · Journal of Visualized Experiments · doi:10.3791/58471 · Cited by 19
Production and Characterization of a High-Affinity Nanobody Against Human Endoglin
Ahmadvand, D., Rasaee, M. J., Rahbarizadeh, F., & Mohammadi, M. (2008) · Hybridoma · doi:10.1089/hyb.2008.0014 · Cited by 20
Nanoantibodies: small molecules, big possibilities
Pedreáñez, A. B., Mosquera-Sulbarán, J., Muñóz, N., Tene, D. M., & Robalino, J. (2021) · Biotechnologia · doi:10.5114/bta.2021.108724 · Cited by 20
di Tommaso, A., Juste, M., Martin‐Eauclaire, M., Dimier‐Poisson, I., Billiald, P., & Aubrey, N. (2012) · Journal of Biological Chemistry · doi:10.1074/jbc.m112.348912 · Cited by 19
Tian, B., Wong, W. Y., Uger, M. D., Wiśniewski, P., & Chao, H. (2017) · Frontiers in Immunology · doi:10.3389/fimmu.2017.00956 · Cited by 19
Current Status of Tissue Clearing and the Path Forward in Neuroscience
Zhao, J., Lai, H. M., Qi, Y., He, D., & Sun, H. (2020) · ACS Chemical Neuroscience · doi:10.1021/acschemneuro.0c00563 · Cited by 18
Development of a platform process for the production and purification of single‐domain antibodies
Crowell, L. E., Goodwine, C., Sosa-Holt, C. S., Rocha, L. A., Vega, C. G., Rodriguez‐Aponte, S. A., Dalvie, N. C., Tracey, M. K., Puntel, M., Wigdorovitz, A., et al. (2021) · Biotechnology and Bioengineering · doi:10.1002/bit.27724 · Cited by 19
“Their small size (12–15 kDa) allows them to bind to previously intractable targets and their increased stability presents the potential for oral dosage (for intestinal diseases) (Harmsen & Haard, 2007).”
BackgroundSelf-Assembled Nanobodies as Selectively Targeted, Nanostructured, and Multivalent Materials
Sánchez‐García, L., Voltà‐Durán, E., Parladé, E., Mazzega, E., Sánchez‐Chardi, A., Serna, N., López‐Laguna, H., Mitstorfer, M., Unzueta, U., Vázquez, E., et al. (2021) · ACS Applied Materials & Interfaces · doi:10.1021/acsami.1c08092 · Cited by 19
Rossotti, M. A., González‐Techera, A., Guarnaschelli, J., Yim, L., Camacho, X., Fernández, M., Cabral, P., Leizagoyen, C., Chabalgoity, J. A., & González‐Sapienza, G. (2015) · mAbs · doi:10.1080/19420862.2015.1068491 · Cited by 19
“These amino acids and a distinctive short CDR3 indicate that T5 derives from a VH(3) segment that recombined with a D-JH encoding a Trp103Arg substitution.(25,26) The DNA sequences of the 3 clones displayed in Figure 1 were cloned in the pINQ-BtH6 vector and the antibody fragments were expressed in the periplasm of E.”
MethodCalpe, S., Wagner, K., Khattabi, M. E., Rutten, L., Zimberlin, C., Dolk, E., Verrips, C. T., Medema, J. P., Spits, H., & Krishnadath, K. K. (2015) · Molecular Cancer Therapeutics · doi:10.1158/1535-7163.mct-14-0956 · Cited by 19
“Their advantageous characteristics have prompted their usage for research, diagnostic, and therapeutic applications (46, 47).”
BackgroundPassive Immunization of Farmed Fish
Rajan, B., Løkka, G., Koppang, E. O., & Austbø, L. (2017) · The Journal of Immunology · doi:10.4049/jimmunol.1700154 · Cited by 21
Wu, D., Yang, J. G., Cong, J., Zhou, J. P., Li, Y., Yan, F., Li, X., Chen, Y., & Ye, L. (2025) · Biosensors and Bioelectronics · doi:10.1016/j.bios.2025.117684 · Cited by 18
Structure and Dynamics Guiding Design of Antibody Therapeutics and Vaccines
Fernández‐Quintero, M. L., Pomarici, N. D., Fischer, A.-L. M., Hoerschinger, V. J., Kroell, K. B., Riccabona, J. R., Kamenik, A. S., Loeffler, J. R., Ferguson, J. A., Perrett, H. R., et al. (2023) · Antibodies · doi:10.3390/antib12040067 · Cited by 21
“These special formats, also known as heavy-chain only antibodies (HcAb), are deprived of the light chains and therefore characterized by the assembly of only two heavy chains [195,196].”
BackgroundEvolution of Therapeutic Antibodies, Influenza Virus Biology, Influenza, and Influenza Immunotherapy
Chaisri, U., & Chaicumpa, W. (2018) · BioMed Research International · doi:10.1155/2018/9747549 · Cited by 18
“They also have high binding affinity to the target [68, 69].”
BackgroundYang, S., Li, L., Yin, S., YouJun, S., Khan, M. U. Z., He, X., Li, Y., Gao, X. M., Liu, X., & Cai, J. (2018) · Applied Microbiology and Biotechnology · doi:10.1007/s00253-018-9324-7 · Cited by 18
Naderi, S., Roshan, R., Ghaderi, H., Behdani, M., Mahmoudi, S., Habibi‐Anbouhi, M., Shokrgozar, M. A., & Kazemi‐Lomedasht, F. (2020) · Molecular Immunology · doi:10.1016/j.molimm.2020.10.004 · Cited by 18
Next Generation Imaging Techniques to Define Immune Topographies in Solid Tumors
Pietrobon, V., Cesano, A., Marincola, F. M., & Kather, J. N. (2021) · Frontiers in Immunology · doi:10.3389/fimmu.2020.604967 · Cited by 18
Nanobody Paratope Ensembles in Solution Characterized by MD Simulations and NMR
Fernández‐Quintero, M. L., DeRose, E. F., Gabel, S. A., Mueller, G. A., & Liedl, K. R. (2022) · International Journal of Molecular Sciences · doi:10.3390/ijms23105419 · Cited by 16
“Nanobodies are potent alternatives to conventional antibodies, because of their small size, refolding capacity, stability, specificity, and natural origin [4,7,8].”
BackgroundTogtema, M., Hussack, G., Dayer, G., Teghtmeyer, M. R., Raphael, S., Tanha, J., & Zehbe, I. (2019) · International Journal of Molecular Sciences · doi:10.3390/ijms20092088 · Cited by 19
Merikhian, P., Darvishi, B., Jalili, N., Esmailinejad, M. R., Khatibi, A. S., Kalbolandi, S. M., Salehi, M., Mosayebzadeh, M., Barough, M. S., Majidzadeh‐A, K., et al. (2021) · Molecular Oncology · doi:10.1002/1878-0261.13123 · Cited by 17
RetractedNguyen, T. D., Bordeau, B. M., Zhang, Y., Mattle, A. G., & Balthasar, J. P. (2022) · International Journal of Molecular Sciences · doi:10.3390/ijms24010475 · Cited by 17
“Furthermore, sdAb isolated from the VHH libraries has several advantages over scFv and Fab, including better stability, higher expression yield, less propensity to form dimers and aggregates, and being smaller and easier to engineer [10,12,32,33].”
BackgroundCalpe, S., Correia, A. C. P., Sancho‐Serra, M. D. C., & Krishnadath, K. K. (2016) · mAbs · doi:10.1080/19420862.2016.1158380 · Cited by 17
“The clinical applicability of VHHs is currently being investigated in a wide array of diseases, such as inflammatory and infectious diseases, as well as oncology.(55) The superiority of VHHs in comparison with conventional antibodies in terms of specificity and potency is well documented.”
BackgroundApproaches to Manipulate Ephrin-A:EphA Forward Signaling Pathway
Baudet, S., Bécret, J., & Nicol, X. (2020) · Pharmaceuticals · doi:10.3390/ph13070140 · Cited by 17
“Finally, nanobodies and scFv can be fused together, creating multimeric nanobodies or scFvs with the ability to recognize multiple epitopes from the same target or from multiple targets for combinatorial effects [94,95].”
BackgroundBitencourt, A. L. B., de Campos, R. M., Cline, E. N., Klein, W. L., & Sebollela, A. S. (2020) · International Journal of Molecular Sciences · doi:10.3390/ijms21238920 · Cited by 18
“Human VH domains and camelid VHH framework regions show a high sequence homology [61].”
BackgroundAbderrazek, R. B., Ksouri, A., Idoudi, F., Dhaouadi, S., Hamdi, E., Vincke, C., Farah, A., Benlasfar, Z., Majdoub, H., Ayeb, M. E., et al. (2022) · Frontiers in Immunology · doi:10.3389/fimmu.2022.863012 · Cited by 18
“, the VL anchoring site in VH) strongly suggested that NbBotI-01 will fail to associate with a VL domain and was therefore considered as a VH-like VHH domain (53, 54) (Figure 2A).”
BackgroundProspective Applications of Single-Domain Antibodies in Biomedicine
Tillib, S. V. (2020) · Molecular Biology · doi:10.1134/s0026893320030164 · Cited by 17
Designed Ankyrin Repeat Proteins as a tool box for analyzing p63
Strubel, A., Münick, P., Chaikuad, A., Dreier, B., Schaefer, J. V., Gebel, J., Osterburg, C., Tuppi, M., Schäfer, B., Knapp, S., et al. (2022) · Cell Death and Differentiation · doi:10.1038/s41418-022-01030-y · Cited by 16
“These shortcomings have been addressed by developing single-chain Fv antibodies (scFvs) that were used to create “ Intrabodies ” without disul fi de bonds [4 – 6].”
MethodSu, S., Esparza, T. J., & Brody, D. L. (2022) · PLoS ONE · doi:10.1371/journal.pone.0276107 · Cited by 16
“The molecular weights of 108 VHHs are typically ~15kDa, which is about 1/10 of the molecular weight of a conventional IgG and about 109 half the molecular weight of a single chain variable fragment (Scfv) [25, 26].”
BackgroundHeidari, M., Shirazi, E. A., Cheraghi, S. F., Shahshahani, R., Rahnama, T., & Khatami, M. (2024) · Molecular Biology Reports · doi:10.1007/s11033-024-09684-2 · Cited by 16
“For tumor therapy, it is essential to humanize nanobod-ies as much as possible without compromising their affinity, expression level, stability, and solubility [46, 47].”
Engineered Antibodies to Improve Efficacy against Neurodegenerative Disorders
Niazi, S. K., Mariam, Z., & Magoola, M. (2024) · International Journal of Molecular Sciences · doi:10.3390/ijms25126683 · Cited by 16
“With a molecular weight of only 12–15 kDa, these are even smaller than Fab fragments (~50 kDa) [134] having a peptide chain of about 110 amino acids long and only one variable domain (VH) of a heavy-chain antibody.”
BackgroundYang, M., Xu, Q., Gu, K., Wen, R., Zhou, C., Zhao, Y. W., Guo, B., Xu, W., Zhang, Y., Li, C., et al. (2024) · Spectrochimica Acta Part A Molecular and Biomolecular Spectroscopy · doi:10.1016/j.saa.2024.125309 · Cited by 19
Metadata
- Authors
- Harmsen, M. M., & De Haard, H. J.
- Affiliations
- Wageningen University & Research (NL)
- Ablynx (Belgium) (BE)
- Corresponding author
- Harmsen, M. M.
- Venue
- Applied Microbiology and Biotechnology
- Published
- 2007-08-18
- Volume
- 77
- Issue
- 1
- Pages
- 13-22
- ISSN
- 0175-7598 (ISSN-L)
- 1432-0614
- PMC
- PMC2039825
- OpenAlex
- W2147451568
- Cited by
- 809
- References
- 96
- Citations by year
- 2026: 31 · 2025: 43 · 2024: 48 · 2023: 41 · 2022: 41 · 2021: 65 · 2020: 57 · 2019: 49 · 2018: 53 · 2017: 42 · 2016: 46 · 2015: 48 · 2014: 53 · 2013: 40 · 2012: 52
- FWCI
- 2.67
- Type
- journal article
- Language
- en
- Open access
- hybrid
- doi.org
- Locations
- Applied Microbiology and Biotechnology (publishedVersion, cc-by)
- PubMed Central (publishedVersion, cc-by)
- PubMed (publishedVersion)
- Socio-Environmental Systems Modeling (acceptedVersion)
- Data Archiving and Networked Services (DANS) (submittedVersion, other-oa)
- Topics
- Life Sciences › Immunology and Microbiology › Immunology › Monoclonal and Polyclonal Antibodies Research
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Protein purification and stability
- Life Sciences › Agricultural and Biological Sciences › Food Science › Animal Diversity and Health Studies
- Keywords
- single-domain antibody fragments, VHH, nanobodies, camelid antibodies, recombinant antibody production, yeast expression system, phage display, ribosome display, yeast surface display, therapeutic applications
- MeSH terms
- Animals, Camelids, New World, Immunoglobulin Fragments, Immunoglobulin Heavy Chains, Models, Biological
About these checks
Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.
Statistics and arithmetic are recomputed, not judged.
Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.
What counts as a claim ›How we measure confidence ›How we check this ›
Are you an author, or do you see a problem with this page? Request a review
