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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.
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“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]
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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).”
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Turner, K. B., Alves, N. J., Medintz, I. L., & Walper, S. A. (2015) · Expert Opinion on Drug Delivery · doi:10.1517/17425247.2016.1133583 · Cited by 16
“165 kDa) results in improved biodistribution, deeper tissue penetration, and reduced serum half-life [18,23] (Figure 2D).”
BackgroundRen, J., Zhang, C., Ji, F., & Jia, L. (2020) · Journal of Chromatography A · doi:10.1016/j.chroma.2020.461227 · Cited by 16
Alkaline Phosphatase‐based Electrochemical Analysis for Point‐of‐Care Testing
Kanno, Y., Zhou, Y., Fukuma, T., & Takahashi, Y. (2021) · Electroanalysis · doi:10.1002/elan.202100294 · Cited by 16
Jiang, H., Xie, Y., Burnette, A., Roach, J. M., Giardina, S. L., Hecht, T. T., Creekmore, S. P., Mitra, G., & Zhu, J. (2012) · Applied Microbiology and Biotechnology · doi:10.1007/s00253-012-4319-2 · Cited by 16
Summanen, M., Granqvist, N., Tuominen, R. K., Yliperttula, M. L., Verrips, C. T., Boonstra, J., Blanchetot, C., & Ekokoski, E. (2012) · PLoS ONE · doi:10.1371/journal.pone.0035630 · Cited by 16
“VHH antibodies generally have affinities comparable to those of conventional antibody fragments, with KD values in the nanomolar range [25], and VHHs with affinity constants as low as 100 pM have been described [24].”
BackgroundA novel nanobody specific for respiratory surfactant protein A has potential for lung targeting
Li, H., Wang, S., He, X., Li, N., Yu, F., Hu, Y., Wang, L., Zhang, P., Du, Y., Du, S.-S., et al. (2015) · International Journal of Nanomedicine · doi:10.2147/ijn.s77268 · Cited by 16
“41,42 To determine further the lung-targeting ability of the recombinant nanobody, we performed in vivo imaging by labeling Nb17 with FITC (FITC-Nb17).”
Affinity Biosensors for Detection of Mycotoxins in Food
Evtugyn, G. A., Šubjaková, V., Melikishvili, S., & Hianik, T. (2018) · Advances in food and nutrition research · doi:10.1016/bs.afnr.2018.03.003 · Cited by 16
Molecular techniques for understanding harmful algal blooms: A review
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A nanobody-based therapeutic targeting Nipah virus limits viral escape
Isaacs, A., Valenzuela, G., Zhang, X., Modhiran, N., Barr, J. A., Thakur, N., Low, Y. S., Parry, R. H., Barnes, J. B., Jara, R., et al. (2025) · Nature Structural & Molecular Biology · doi:10.1038/s41594-025-01598-2 · Cited by 18
“41,42 .”
Hennigan, J. N., Menacho‐Melgar, R., Sarkar, P., Golovsky, M., & Lynch, M. (2023) · Metabolic Engineering · doi:10.1016/j.ymben.2024.07.012 · Cited by 17
Generation of VHH antibodies against the Arabidopsis thaliana seed storage proteins
Meyer, T. D., Eeckhout, D., Rycke, R. M. D., Buck, S. D., Muyldermans, S., & Depicker, A. (2013) · Plant Molecular Biology · doi:10.1007/s11103-013-0118-0 · Cited by 15
Bakherad, H., Gargari, S. L. M., Sepehrizadeh, Z., Aghamollaei, H., Taheri, R. A., Torshabi, M., Yazdi, M. T., Ebrahimizadeh, W., & Setayesh, N. (2017) · Biomedicine & Pharmacotherapy · doi:10.1016/j.biopha.2017.06.025 · Cited by 15
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Tillib, S. V. (2020) · Molecular Biology · doi:10.31857/s0026898420030167 · Cited by 15
Creation of stable water-free antibody based protein liquids
Slocik, J. M., Dennis, P. B., Kuang, Z., Pelton, A. T., & Naik, R. R. (2021) · Communications Materials · doi:10.1038/s43246-021-00222-2 · Cited by 15
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Domingo‐Espín, J., Unzueta, U., Saccardo, P., Rodríguez‐Carmona, E., Corchero, J. L., Vázquez, E., & Ferrer‐Miralles, N. (2011) · Progress in molecular biology and translational science · doi:10.1016/b978-0-12-416020-0.00006-1 · Cited by 17
Kobayashi, N., Oyama, H., Nakano, M., Kanda, T., Banzono, E., Katō, Y., Karibe, T., Nishio, T., & Goto, J. (2009) · Analytical Biochemistry · doi:10.1016/j.ab.2009.01.004 · Cited by 15
Jiang, W., Rosenberg, J. N., Wauchope, A. D., Tremblay, J. M., Shoemaker, C. B., Weeks, D. P., & Oyler, G. A. (2013) · The Plant Journal · doi:10.1111/tpj.12316 · Cited by 15
Mu, X., Tong, Z., Huang, Q., Liu, B., Liu, Z., Lan-Qun, H. A. O., Hua, D., Zhang, J., & Gao, C. (2016) · Sensors · doi:10.3390/s16030308 · Cited by 15
Kobayashi, S., Terai, T., Yoshikawa, Y., Ohkawa, R., Ebihara, M., Hayashi, M., Takiguchi, K., & Nemoto, N. (2017) · Chemical Communications · doi:10.1039/c7cc00099e · Cited by 15
Vostakolaei, M. A., Molavi, O., Hejazi, M. S., Kordi, S., Rahmati, S., Barzegari, A., & Abdolalizadeh, J. (2019) · Journal of Cellular Biochemistry · doi:10.1002/jcb.28732 · Cited by 15
“Antibody fragments such as single‐chain fragments variable (scFvs) can be produced using PDT, while the cost of production is low than other antibody productions methods and can show the same binding specificity and affinity as the monomeric form of the full antibody always.(32) The scFvs fragments comprised a single…”
BackgroundNanobody-Functionalized Cellulose for Capturing SARS-CoV-2
Sun, X., Yang, S., Al‐Dossary, A. A., Broitman, S., Ni, Y., Guan, M., Yang, M., & Li, J. (2022) · Applied and Environmental Microbiology · doi:10.1128/aem.02303-21 · Cited by 14
“mammalian cell hosts for production (24).”
Dorresteijn, B., Rotman, M., Faber, D., Schravesande, R., Suidgeest, E., van der Weerd, L., van der Maarel, S. M., Verrips, C. T., & Khattabi, M. E. (2015) · FEBS Journal · doi:10.1111/febs.13367 · Cited by 15
Optimised nanobody-based quenchbodies for enhanced protein detection
Cater, J. H., Salamouni, N. S. E., Mansour, G. H., Hutchinson, S., Guinness, C. M., Mueller, S. H., Spinks, R. R., Shanmugam, N., Pichard-Kostuch, A., Zahoransky, V., et al. (2025) · Communications Biology · doi:10.1038/s42003-025-08359-3 · Cited by 14
“Respectfully, we disagree with their comment on the improvement of the quenchbody performance and believe this may be a misunderstanding on behalf of reviewer #3.”
Emerging monoclonal antibodies againstClostridium difficileinfection
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Single molecule imaging of protein aggregation in Dementia: Methods, insights and prospects
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Harmsen, M. M., Ackerschott, B., & de Smit, H. (2024) · Frontiers in Immunology · doi:10.3389/fimmu.2024.1346328 · Cited by 14
Self-citationMa, J., Tian, Z., Shi, Q., Dong, X., & Sun, Y. P. (2024) · Journal of Chromatography A · doi:10.1016/j.chroma.2024.464851 · Cited by 14
Cuypers, M., Jaspers, T., Clerckx, J., Leekens, S., Cawthorne, C., Bormans, G. M., Cleeren, F., Geukens, N., Strooper, B. D., & Dewilde, M. (2025) · Fluids and Barriers of the CNS · doi:10.1186/s12987-025-00624-1 · Cited by 15
Mu, X., Tong, Z., Huang, Q., Liu, B., Liu, Z., Lan-Qun, H. A. O., Zhang, J., Gao, C., & Wang, F. (2015) · Sensors · doi:10.3390/s150203896 · Cited by 14
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Veugelen, S., Dewilde, M., Strooper, B. D., & Chávez‐Gutiérrez, L. (2016) · Methods in enzymology on CD-ROM/Methods in enzymology · doi:10.1016/bs.mie.2016.10.029 · Cited by 15
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“In contrast to intact antibodies, nanobodies have been proved to be very efficiently produced in microbial hosts including Escherichia coli,([11]) Pichia pastoris,([12]) and Saccharomyces cerevisiae.”
BackgroundKane, J., Jansen, M. F., Hendrix, S., Bosmans, L. A., Beckers, L., van Tiel, C. M., Gijbels, M. J., Zelcer, N., de Vries, C. J. M., von Hundelshausen, P., et al. (2021) · Thrombosis and Haemostasis · doi:10.1055/a-1711-1055 · Cited by 14
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Buonocore, M., Grimaldi, M., Santoro, A., Covelli, V., Marino, C., Napolitano, E., Novi, S., Tecce, M. F., Ciaglia, E., Montella, F., et al. (2023) · International Journal of Molecular Sciences · doi:10.3390/ijms242115610 · Cited by 14
Selection and characterization of camelid nanobodies towards urokinase-type plasminogen activator
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Inhibition of glycosylation on a camelid antibody uniquely affects its FcγRI binding activity
Krahn, N., Spearman, M. A., Meier, M., Dorion-Thibaudeau, J., McDougall, M., Patel, T. R., Crescenzo, G. D., Durocher, Y., Stetefeld, J., & Butler, M. (2016) · European Journal of Pharmaceutical Sciences · doi:10.1016/j.ejps.2016.09.040 · Cited by 14
Liu, Z., Huang, D., Fu, X., Peng, C., & Du, E. (2018) · Biotechnology & Biotechnological Equipment · doi:10.1080/13102818.2017.1420426 · Cited by 14
“coli cytoplasm with a yield of 1–10 mg per liter bacterial culture [28].”
BackgroundCombinatorial Design of a Nanobody that Specifically Targets Structured RNAs
Cawez, F., Duray, E., Hu, Y., Vandenameele, J., Romão, E., Vincke, C., Dumoulin, M., Galleni, M., Muyldermans, S., & Vandevenne, M. S. (2018) · Journal of Molecular Biology · doi:10.1016/j.jmb.2018.03.032 · Cited by 14
“In addition, nanobodies are more soluble and stable than other antigen-binding domains [28].”
BackgroundSadeghian-Rizi, T., Behdani, M., Khanahmad, H., Hamid, M., & Jahanian‐Najafabadi, A. (2018) · CNS & Neurological Disorders - Drug Targets · doi:10.2174/1871527317666181114134518 · Cited by 14
Stengl, A., Gerlach, M., Kasper, M., Hackenberger, C. P. R., Leonhardt, H., Schumacher, D., & Helma, J. (2019) · Organic & Biomolecular Chemistry · doi:10.1039/c9ob00508k · Cited by 14
Kumar, S., Mahendran, I., Athreya, A., Ranjan, R. D., & Penmatsa, A. (2019) · Journal of Biological Chemistry · doi:10.1074/jbc.ra119.010902 · Cited by 13
Park, B. G., Kim, Y. J., Min, J. H., Cheong, T.-C., Nam, S. H., Cho, N., Kim, Y. K., & Lee, K. B. (2020) · Nanoscale Research Letters · doi:10.1186/s11671-020-03395-w · Cited by 14
Schneider, F., Sych, T., Eggeling, C., & Sezgin, E. (2020) · iScience · doi:10.1016/j.isci.2020.101891 · Cited by 14
Harmsen, M. M., Blokker, J., Pritz-Verschuren, S. B. E., Bartelink, W., der Burg, H. V., & Koch, G. (2013) · Antibodies · doi:10.3390/antib2020168 · Cited by 14
Self-citationBlank, L., Pander, G., Mühlberg, E., Mier, W., & Uhl, P. (2025) · Drug Discovery Today · doi:10.1016/j.drudis.2025.104524 · Cited by 15
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“However, they have some drawbacks such as little permeability to tumor tissues, low stability and their expensive costs [15].”
BackgroundEvolving therapeutic proteins to precisely kill cancer cells
Zaman, R., Islam, R. A., & Chowdhury, E. H. (2022) · Journal of Controlled Release · doi:10.1016/j.jconrel.2022.09.066 · Cited by 13
Isolation of a Novel Nanobody Against HER-2/neuUsing Phage Displays Technology
Sheikholeslami, F., Rasaee, M. J., Shokrgozar, M. A., Dizaji, M. M., Rahbarizadeh, F., & Ahmadvande, D. (2010) · Laboratory Medicine · doi:10.1309/lm0wxkm0r0dvuzwf · Cited by 13
Srivastava, S. K., Ruigrok, V. J. B., Thompson, N., Trilling, A. K., Heck, A. J. R., van Rijn, C. J. M., Beekwilder, J., & Jongsma, M. A. (2013) · PLoS ONE · doi:10.1371/journal.pone.0064040 · Cited by 13
“Furthermore, they can be produced at low cost in yeast or bacteria, and are easy to handle with long shelf life [16,17].”
Selection of Antibody Fragments by Yeast Display
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“VHHs have been used for in vivo imaging and therapeutics [25,26].”
BackgroundMethodWang, D., Yang, S., Yin, S., YouJun, S., Du, P., Guo, J., He, J., Cai, J., & Liu, X. (2015) · BMC Veterinary Research · doi:10.1186/s12917-015-0437-2 · Cited by 13
“However, sdAbs derived from nonimmune libraries show low affinities for their target antigens [7, 38, 39].”
BackgroundJi, X., Lu, W., Zhou, H., Han, D., Yang, L., Wu, H., Li, J., Liu, H., Zhang, J., Cao, P., et al. (2013) · Applied Microbiology and Biotechnology · doi:10.1007/s00253-012-4639-2 · Cited by 13
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“134–137 They are cost-effective to manufacture and exhibit superior binding affinity and specificity against various targets, including tumor markers.”
Nanobodies for the Early Detection of Ovarian Cancer
Tran, L., Graulus, G., Vincke, C., Smiejkowska, N., Kindt, A., Devoogdt, N., Muyldermans, S., Adriaensens, P., & Guedens, W. (2022) · International Journal of Molecular Sciences · doi:10.3390/ijms232213687 · Cited by 12
Cai, C., Liu, Y., Tang, X., Zhang, W., Zhang, Q., & Li, P. (2023) · LWT · doi:10.1016/j.lwt.2023.114829 · Cited by 12
Rafique, A., Satake, K., Kishimoto, S., Khan, K. H., Kato, D., & Ito, Y. (2019) · Monoclonal Antibodies in Immunodiagnosis and Immunotherapy · doi:10.1089/mab.2019.0027 · Cited by 12
“(21,22) There are two types of HC Ab, classified as IgG 2 and IgG 3 , which have short and long hinge regions, respectively, between variable antigen-binding domain VHH and Fc.”
Kumada, Y., Kang, B., Yamakawa, K., Kishimoto, M., & Horiuchi, J. (2015) · Biotechnology Progress · doi:10.1002/btpr.2169 · Cited by 12
“ubility in a solution due to a simple beta strand structure with an intramolecular disulfide bridge.(3) For these reasons,”
BackgroundAffinity maturation of a CDR3-grafted VHH using in silico analysis and surface plasmon resonance
Inoue, H., Suganami, A., Ishida, I., Tamura, Y., & Maeda, Y. (2013) · The Journal of Biochemistry · doi:10.1093/jb/mvt058 · Cited by 12
Suzuki, R., Saito, K., Matsuda, M., Sato, M., Kanegae, Y., Shi, G., Watashi, K., Aizaki, H., Chiba, J., Saito, I., et al. (2016) · Journal of General Virology · doi:10.1099/jgv.0.000423 · Cited by 12
Prevention of Vγ9Vδ2 T Cell Activation by a Vγ9Vδ2 TCR Nanobody
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“Furthermore, owing to the single-domain character and small size (∼15 kDa) of VHHs, they have additional advantages, including enhanced tissue/tumor penetration, enhanced stability and solubility, and ease of production in relatively cost- and time-efficient production systems such as bacteria or yeast (27, 63).”
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He, Q., Pan, J., Zhang, Y., Xu, Z., Morisseau, C. H. P., Hammock, B. D., & Li, D. (2025) · TrAC Trends in Analytical Chemistry · doi:10.1016/j.trac.2025.118404 · Cited by 11
Liu, X., Wang, C., Bai, Y., Wang, W., Han, Y., Cai, S., An, J., & Qu, G. (2024) · Microbial Pathogenesis · doi:10.1016/j.micpath.2024.106615 · Cited by 11
Analysis of EGFR binding hotspots for design of new EGFR inhibitory biologics
Tydings, C. W., Singh, B., Smith, A. W., Ledwitch, K. V., Brown, B. P., Lovly, C. M., Walker, A. S., & Meiler, J. (2024) · Protein Science · doi:10.1002/pro.5141 · Cited by 13
Damsbo, A., Burlet, N. J., Fernández‐Quintero, M. L., Bénard-Valle, M., Overath, M. D., Guadarrama-Martínez, A., Vlamynck, A., Bisbo, I., Villalobos, C., Tulika, T., et al. (2025) · Communications Chemistry · doi:10.1038/s42004-025-01600-4 · Cited by 13
“V H Hs have demonstrated a generally stable, monomeric, and aggregation-resistant behaviour 18,19 .”
Yin, S., Yang, S., YouJun, S., Sun, S., Zhou, G., Ye, J., Tian, H. W., Wu, J., & Liu, X. (2013) · PLoS ONE · doi:10.1371/journal.pone.0063500 · Cited by 11
Optical sensors of heterotrimeric G protein signaling
Bondar, A., & Lazar, J. (2020) · FEBS Journal · doi:10.1111/febs.15655 · Cited by 9
Site-Specific Modification of Single Domain Antibodies by Enzyme-Immobilized Magnetic Beads
Zhou, S., Fang, X., Lv, J., Yang, Y., Zeng, Y., Liu, Y., Wei, W., Huang, G., Zhang, B., & Wu, C. (2023) · Bioconjugate Chemistry · doi:10.1021/acs.bioconjchem.3c00423 · Cited by 11
The Application of Prophylactic Antibodies for Rhinovirus Infections
Privolizzi, R., Solari, R. C. E., Johnston, S. L., & McLean, G. R. (2013) · Antiviral chemistry & chemotherapy · doi:10.3851/imp2578 · Cited by 11
Pregnancy-Associated Changes of IgG and Serum N-Glycosylation in Camel (Camelus dromedarius)
Adamczyk, B., Albrecht, S., Stöckmann, H., Ghoneim, I. M., Al-Eknah, M. M., Al-Busadah, K. A., Karlsson, N. G., Carrington, S. D., & Rudd, P. M. (2016) · Journal of Proteome Research · doi:10.1021/acs.jproteome.6b00439 · Cited by 11
Hollander, M. R., Jansen, M. F., Hopman, L. H. G. A., Dolk, E., van de Ven, P. M., Knaapen, P., Horrevoets, A. J. G., Lutgens, E., & van Royen, N. (2019) · Journal of the American Heart Association · doi:10.1161/jaha.119.012806 · Cited by 11
“Daily administration was chosen because of the relatively high blood clearance of VHHs.(14) The charging dose consisted of either 0.”
BackgroundMethodInfluentialRecent progress in antitumor functions of the intracellular antibodies
Lin, Y., Chen, Z., Hu, C., Chen, Z., & Zhang, L. (2020) · Drug Discovery Today · doi:10.1016/j.drudis.2020.02.009 · Cited by 11
Tissue-specific targeting of DNA nanodevices in a multicellular living organism
Chakraborty, K., Anees, P., Surana, S., Martin, S., Aburas, J., Moutel, S., Perez, F., Koushika, S. P., Kratsios, P., & Krishnan, Y. (2021) · eLife · doi:10.7554/elife.67830 · Cited by 11
Wang, K., Yu, Y., Xu, Y., Yue, Y., Zhao, F., Feng, W., Duan, Y., Duan, W., Yue, J., Liao, Z., et al. (2023) · Cell & Bioscience · doi:10.1186/s13578-023-01043-1 · Cited by 10
“Alternatively, nanobodies could be used, which are single-domain antibody fragments with a molecular weight of only 12–15 kDa [43].”
BackgroundSingle-domain antibodies and aptamers drive new opportunities for neurodegenerative disease research
Shoemaker, R. L., Larsen, R. J., & Larsen, P. A. (2024) · Frontiers in Immunology · doi:10.3389/fimmu.2024.1426656 · Cited by 10
Genetic Removal of the CH1 Exon Enables the Production of Heavy Chain-Only IgG in Mice
Zhang, T., Cheng, X., Yu, D., Lin, F., Hou, N., Cheng, X., Hao, S., Wei, J., Ma, L., Fu, Y., et al. (2018) · Frontiers in Immunology · doi:10.3389/fimmu.2018.02202 · Cited by 10
“Since their discovery more than 25 years ago, nano-antibodies have been used for research purposes, small chemical analyses, clinical diagnosis, and therapeutic applications (1, 12– 16), due to their apparent advantageous characteristics, such as a small size and better solubility and stability than conventional antib…”
BackgroundMacromolecules and Antibody-Based Drugs
Su, X., & Yao, S. (2020) · Advances in experimental medicine and biology · doi:10.1007/978-981-15-3266-5_20 · Cited by 11
Introduction of an Aldehyde Handle on Nanobodies by Affinity-Guided Labeling
Mortensen, M. R., Skovsgaard, M. B., Märcher, A., Andersen, V. L., Palmfeldt, J., Nielsen, T. B., Tørring, T., Laursen, N. S., Andersen, K. R., Kjems, J., et al. (2020) · Bioconjugate Chemistry · doi:10.1021/acs.bioconjchem.0c00151 · Cited by 10
Llama DNA Immunization and Isolation of Functional Single-Domain Antibody Binders
Trempe, F., Rossotti, M. A., Maqbool, T., MacKenzie, C. R., & Arbabi‐Ghahroudi, M. (2022) · Methods in molecular biology · doi:10.1007/978-1-0716-2075-5_3 · Cited by 10
Total Chemical Synthesis of a Functionalized GFP Nanobody
Huppelschoten, Y., Hebieshy, A. F. E., Hameed, D. S., Sapmaz, A., Buchardt, J., Nielsen, T. E., Ovaa, H., & van Noort, G. J. V. D. H. (2022) · ChemBioChem · doi:10.1002/cbic.202200304 · Cited by 10
Lim, H. T., Kok, B. H., Lim, C. P., Majeed, A. B. A., Leow, C. Y., & Leow, C. Y. (2022) · Biomedical Engineering Advances · doi:10.1016/j.bea.2022.100054 · Cited by 10
Radiotherapy plus CAR-T cell therapy to date: A note for cautions optimism?
Tian, H., Li, H., & Tang, B. (2022) · Frontiers in Immunology · doi:10.3389/fimmu.2022.1033512 · Cited by 10
“Nanobodies also tend to have an advantage over single-chain antibodies in the context of humanization due to the more straightforward humanization process of nanobodies (77).”
BackgroundGuo, Y., Kamara, S., Zhang, J., He, W., Zheng, M., Liu, Y., Zhou, L., Chen, J., Zhu, S., & Zhang, L. (2023) · Frontiers in Cellular and Infection Microbiology · doi:10.3389/fcimb.2022.1078504 · Cited by 10
Production, expression, and function of dual-specific monoclonal antibodies in a single plant
Jin, C., Kang, Y. J., Park, S. R., Oh, Y. J., & Ko, K. (2023) · Planta · doi:10.1007/s00425-023-04284-z · Cited by 10
“Such antibodies come in a variety of forms, including full size, ScFv, nanobody, large single chain (LSC), and camelid antibodies, and have been modified to express in heterologous biosystems (Harmsen and De Haard 2007).”
BackgroundFranceschi, V., Jacca, S., Sassu, E. L., Stellari, F. F., van Santen, V. L., & Donofrío, G. (2014) · PLoS ONE · doi:10.1371/journal.pone.0105643 · Cited by 10
“To stabilize the enlarged CDRs, nanobodies often possess an additional disulfide bond between CDR1 and CDR3 in dromedaries, and CDR2 and CDR3 in llamas and alpaca [33].”
Generation and characterization of nanobodies against rhGH expressed as sfGFP fusion protein
Abbady, A. Q., Al-Shemali, R., Assaad, J. M., & Murad, H. (2014) · General and Comparative Endocrinology · doi:10.1016/j.ygcen.2014.05.018 · Cited by 10
Brower, K., Ryakala, V. K., Bird, R., Godawat, R., Riske, F. J., Konstantinov, K., Warikoo, V., & Gamble, J. (2014) · Biotechnology Progress · doi:10.1002/btpr.1870 · Cited by 10
Kieninger, B., Владиславовна, Г. З., Krüger, S., Westermark, G. T., Friedrich, R. P., Fändrich, M., & Röcken, C. (2011) · Amyloid · doi:10.3109/13506129.2011.560623 · Cited by 10
The current perspectives of dromedary camel stem cells research
Saadeldin, I. M., Swelum, A. A., Alzahrani, F. A., & Alowaimer, A. N. (2018) · International Journal of Veterinary Science and Medicine · doi:10.1016/j.ijvsm.2018.01.002 · Cited by 10
“Owing to their very small size, they are able to penetrate the typically inaccessible parts of the solid tumors with low immunogenicity [26].”
BackgroundRecent Advances in the Scaffold Engineering of Protein Binders
Ahmadi, M. K. B., Mohammadi, S. A., Makvandi, M., mamouei, M., Rahmati, M., Dehghani, H., & Wood, D. W. (2020) · Current Pharmaceutical Biotechnology · doi:10.2174/1389201021999200824101035 · Cited by 10
99mTc-labeled single-domain antibody EG2 in targeting epidermal growth factor receptor
Li, C., Wen, B., Wang, L., Feng, H., Xia, X., Ding, Z., Gao, B., Zhang, Y., & Lan, X. (2015) · Nuclear Medicine Communications · doi:10.1097/mnm.0000000000000264 · Cited by 10
Isolation of bovine serum albumin from whey using affinity chromatography
Besselink, T., Janssen, A. E. M., & Boom, R. M. (2014) · International Dairy Journal · doi:10.1016/j.idairyj.2014.09.004 · Cited by 10
Su, J., Beh, C. W., Ohmuro‐Matsuyama, Y., Kitaguchi, T., Hoon, S. S., & Ueda, H. (2019) · Journal of Bioscience and Bioengineering · doi:10.1016/j.jbiosc.2019.05.015 · Cited by 10
Antibody fragments as therapeutics
(2012) · Elsevier eBooks · doi:10.1533/9781908818096.265 · Cited by 10
Tanaka, Y., Miyazaki, Y., Kawanishi, M., Yamasaki, H., & Takeuchi, T. (2024) · RMD Open · doi:10.1136/rmdopen-2024-004480 · Cited by 9
A Nanobody of PEDV S1 Protein: Screening and Expression in Escherichia coli
Hao, Z., Dong, X., Zhang, Z., & Qin, Z. (2024) · Biomolecules · doi:10.3390/biom14091116 · Cited by 10
“These positions are conserved noncanonical VH structural domains [30], and these four amino acids are hydrophilic, making the surface of Nb more hydrophilic, thereby increasing its water solubility [31].”
BackgroundYuen, D., Feeney, O. M., Noi, L., Shengule, S. R., McLeod, V. M., Reitano, P., Tsegay, S., Hufton, R., Houston, Z. H., Fletcher, N. L., et al. (2025) · ACS Nano · doi:10.1021/acsnano.4c10851 · Cited by 9
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
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