Construction of multiform scFv antibodies using linker peptide
Abstract
Multiform single chain variable fragments (scFvs) including different length linker scFvs and bispecific scFv were constructed. The linker lengths of 0, 3, 5, 8, 12, and 15 amino acids between V(H) and V(L) of antideoxynivalenol (anti-DON) scFv were used to analyze the affinities of scFvs. The affinity constants of these scFvs increased when the linker was lower than 12 amino acids. The affinity constant would not change when the linker was longer than 12 amino acids. Fusion gene of anti-DON scFv and antizearalenone (anti-ZEN) scFv was also constructed through connection by a short peptide linker DNA to express a bispecific scFv. The affinity constants assay showed that the two scFvs of fusion bispecific scFv remained their own affinity compared to their parental scFvs. Competitive direct enzyme linked immunosorbent assay was used to detect DON and ZEN in contaminated wheat (Triticum aestivum L.) samples, and the results indicated that this bispecific scFv was applicable in DON and ZEN detection. This work confirmed that bispecific scFv could be successfully obtained, and might also have an application in diagnosing fungal infection, and breeding transgenic plants.
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Linker length influences oligomeric state: linkers greater than 15 residues generally lead to monomers, linkers of 6-15 residues can favor stable dimers and trimers, and linkers with fewer than five residues result in higher-order multimeric molecules.
“It has been largely demonstrated that the linker length influences the oligomeric state. Linkers greater than 15 residues generally lead to monomers while linkers of 6–15 residues can be utilized to deliberately favor the formation of stable dimers and trimers [17,18]. Linkers with fewer than five residues result in the generation of higher-order multimeric molecules [19,20].”
From Evolution of Escherichia coli Expression System in Producing Antibody Recombinant Fragments (2020) · cites this paper as [20]
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Evolution of Escherichia coli Expression System in Producing Antibody Recombinant Fragments
Sandomenico, A., Sivaccumar, J. P., & Ruvo, M. (2020) · International Journal of Molecular Sciences · doi:10.3390/ijms21176324 · Cited by 134 · see citing works
“Additionally, the Lemo systemTM [20], where the expression is under the tight control of the T7 RNAP activity and the target gene expression level is modulated by l-rhamnose, has been used for the production of soluble and properly folded scFv fragments [69,70].”
BackgroundMethod
- Authors
- Wang, S., Zheng, C., Liu, Y., Zheng, H., & Wang, Z.
- Affiliations
- Fujian Agriculture and Forestry University (CN)
- Venue
- Journal of Genetics and Genomics
- Publisher
- Elsevier BV
- Published
- 2008-05
- Volume
- 35
- Issue
- 5
- Pages
- 313-316
- ISSN
- 1873-5533 (ISSN-L)
- 1673-8527
- OpenAlex
- W2020362561
- Cited by
- 20
- References
- 0
- Citations by year
- 2024: 1 · 2022: 1 · 2020: 2 · 2019: 4 · 2017: 2 · 2016: 3 · 2015: 2 · 2013: 1 · 2012: 1
- FWCI
- 0.77
- Type
- journal article
- Language
- en
- Open access
- closed
- Locations
- Journal of genetics and genomics/Journal of Genetics and Genomics (publishedVersion)
- PubMed (publishedVersion)
- Topics
- Health Sciences › Medicine › Radiology, Nuclear Medicine and Imaging › Monoclonal and Polyclonal Antibodies Research
- Life Sciences › Agricultural and Biological Sciences › Plant Science › Mycotoxins in Agriculture and Food
- Life Sciences › Immunology and Microbiology › Immunology › Toxin Mechanisms and Immunotoxins
- Keywords
- scFv, antibody affinity, fusion genes, mycotoxin detection, deoxynivalenol, wheat, transgenic plants, fungal infection diagnosis
- MeSH terms
- Antibodies, Antibody Affinity, Base Sequence, Cloning, Molecular, Enzyme-Linked Immunosorbent Assay, Escherichia coli, Food Contamination, Immunoglobulin Variable Region, Peptides, Trichothecenes, Triticum, Zearalenone, Gene Expression, Antibodies, Bispecific
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