Comparison of physical chemical properties of llama VHH antibody fragments and mouse monoclonal antibodies
Abstract
Antigen specific llama VHH antibody fragments were compared to antigen specific mouse monoclonal antibodies with respect to specificity, affinity and stability. The llama VHH antibody fragments and the mouse monoclonal antibodies investigated were shown to be highly specific for the protein antigen hCG or the hapten antigen RR-6. The affinity of the interaction between monovalent llama VHH antibody fragments and their antigen is close to the nanomolar range, similar to the bivalent mouse monoclonal antibodies studied. Llama VHH antibody fragments are similar to mouse monoclonal antibodies with respect to antigen binding in the presence of ammonium thiocyanate and ethanol. The results show that relative to antigen specific mouse monoclonal antibodies, antigen specific llama VHH fragments are extremely temperature stable. Two out of six llama VHHs are able to bind antigen specifically at temperatures as high as 90 degrees C, whereas four out of four mouse monoclonal antibodies are not functional at this temperature. Together with the finding that llama VHH fragments can be produced at high yield in Saccharomyces cerevisiae, these findings indicate that in the near future antigen specific llama VHH fragments can be used in for antibodies unexpected products and processes.
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VNAs offer some advantages over classical antibodies such as ease to engineer, improved heat and pH stability, and better tissue penetration.
“In general, VNAs offer some advantages over classical antibodies such as ease to engineer, improved heat and pH stability, and better tissue penetration (Hamers‐Casterman et al, 1993; van der Linden et al, 1999; Tillib et al, 2013).”
From mRNA mediates passive vaccination against infectious agents, toxins, and tumors (2017) · cites this paper as van der Linden et al, 1999
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“Contrary to conventional antibodies, VHHs have been shown to remain functional at 90°C (Van der Linden et al. 1999) or after incubation at high temperatures (Van der Linden et al. 1999; Perez et al. 2001).”
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- Authors
- van der Linden, R. H. J., Frenken, L. G. J., de Geus, B., Harmsen, M. M., Ruuls, R. C., Stok, W., de Ron, L., Wilson, S., Davis, P., & Verrips, C. T.
- Affiliations
- Unilever Research and Development Vlaardingen (NL)
- Unilever (United Kingdom) (GB)
- Utrecht University (NL)
- Venue
- Biochimica et Biophysica Acta (BBA) - Protein Structure and Molecular Enzymology
- Publisher
- Elsevier BV
- Published
- 1999-04
- Volume
- 1431
- Issue
- 1
- Pages
- 37-46
- ISSN
- 0167-4838 (ISSN-L)
- OpenAlex
- W1976784576
- Cited by
- 471
- References
- 27
- Citations by year
- 2026: 21 · 2025: 20 · 2024: 21 · 2023: 20 · 2022: 27 · 2021: 29 · 2020: 26 · 2019: 16 · 2018: 17 · 2017: 25 · 2016: 17 · 2015: 18 · 2014: 17 · 2013: 24 · 2012: 20
- FWCI
- 8.00
- Type
- journal article
- Language
- en
- Open access
- closed
- Locations
- Biochimica et Biophysica Acta (BBA) - Protein Structure and Molecular Enzymology (publishedVersion)
- PubMed (publishedVersion)
- Topics
- Health Sciences › Medicine › Radiology, Nuclear Medicine and Imaging › Monoclonal and Polyclonal Antibodies Research
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Protein purification and stability
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › vaccines and immunoinformatics approaches
- Keywords
- nanobodies, mouse monoclonal antibodies, antibody specificity, antigen binding affinity, thermal stability, human chorionic gonadotropin, Saccharomyces cerevisiae expression, therapeutic antibodies
- MeSH terms
- Ethanol, Animals, Antibodies, Monoclonal, Antibody Specificity, Camelids, New World, Enzyme-Linked Immunosorbent Assay, Immunoglobulin Fragments, Immunoglobulin Variable Region, Immunoglobulin Heavy Chains, Temperature, Thiocyanates, Mice
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