Isolation of single-chain antibody fragments against Venezuelan equine encephalomyelitis virus from two different immune sources.
Abstract: Venezuelan equine encephalomyelitis (VEE) virus is an important human and veterinary pathogen of Central and South America. The virus can cause widespread epidemics, affecting hundreds of thousands of horses, and thousands of humans. Detection of the virus early in infection and in mosquito populations may allow epidemics to be predicted such that suitable prophylaxis, such as vaccination, can be used to reduce disease severity and transmission. The sensitivity and specificity of current immunoassays, based on conventional monoclonal and polyclonal antibodies, needs to be improved for the diagnosis of infection. We have examined phage display libraries expressing single-chain antibodies (scFv) produced from two different immune sources, a hybridoma cell line and an immunized mouse spleen. The libraries were panned against VEE virus to select for specific scFvs. scFvs isolated from both libraries were specific for the same epitope on the VEE virus and sequence analysis showed that the scFvs were almost identical apart from the CDR3 region of the heavy chain. The data presented in this article suggest that although scFvs may be useful tools for the detection of viruses, there are serious limitations with the use of phage display as a tool for the isolation of specific antibodies.
Publication Date: 2001-09-27 PubMed ID: 11572636DOI: 10.1089/088282401753266774Google Scholar: Lookup
The Equine Research Bank provides access to a large database of publicly available scientific literature. Inclusion in the Research Bank does not imply endorsement of study methods or findings by Mad Barn.
- Evaluation Study
- Journal Article
Summary
This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.
The study focuses on the development of a more specific and sensitive method to detect Venezuelan equine encephalomyelitis virus (VEE) by isolating single-chain antibodies from two immune sources.
Background of the Study
- The VEE virus is a significant human and veterinary microorganism in Central and South America, capable of causing mass epidemics impacting horses and humans alike.
- Early detection of this virus in the infection phase or in mosquito populations could potentially avert epidemics, paving the way for efficient prophylactic measures such as vaccination to control the disease spread and severity.
- Due to the need for improved diagnostic sensitivity, this research aims to enhance currently available immunoassays using conventional polyclonal and monoclonal antibodies.
Research Method
- The research team focused on phage display libraries with single-chain antibodies (scFv) generated from two different immune sources – an immunized mouse spleen and a hybridoma cell line.
- These libraries underwent panning against the VEE virus to isolate specific scFvs.
Results and Findings
- The scFvs isolated from both libraries were found to target the same epitope on the VEE virus, indicating specificity. A sequence analysis revealed that the scFvs were almost identical except for the CDR3 region of the heavy chain.
- The findings suggest that scFvs can provide a useful detection tool for viruses. However, there are notable restrictions with the use of phage display as an isolation tool for specific antibodies, calling into question the method’s efficiency.
Conclusions
- The research concludes that while promising, the method of using scFvs from two distinct immune sources and phage display libraries still has significant limitations. Further work is required to mitigate these limitations and improve the effectiveness of this diagnostic approach for the VEE virus.
Cite This Article
APA
Duggan JM, Coates DM, Ulaeto DO.
(2001).
Isolation of single-chain antibody fragments against Venezuelan equine encephalomyelitis virus from two different immune sources.
Viral Immunol, 14(3), 263-273.
https://doi.org/10.1089/088282401753266774 Publication
Researcher Affiliations
- Biomedical Sciences, DERA, Porton Down, Wiltshire, UK. jmduggan@dstl.gov.uk
MeSH Terms
- Amino Acid Sequence
- Animals
- Antigens, Viral / immunology
- Cell Line
- Encephalitis Virus, Venezuelan Equine / genetics
- Encephalitis Virus, Venezuelan Equine / immunology
- Encephalomyelitis, Venezuelan Equine / diagnosis
- Encephalomyelitis, Venezuelan Equine / immunology
- Encephalomyelitis, Venezuelan Equine / virology
- Enzyme-Linked Immunosorbent Assay
- Humans
- Hybridomas / immunology
- Immunoglobulin Fragments / immunology
- Immunoglobulin Fragments / isolation & purification
- Mice
- Molecular Sequence Data
- Peptide Library
- Spleen / cytology
- Spleen / immunology
Citations
This article has been cited 9 times.- Roth KDR, Wenzel EV, Ruschig M, Steinke S, Langreder N, Heine PA, Schneider KT, Ballmann R, Fühner V, Kuhn P, Schirrmann T, Frenzel A, Dübel S, Schubert M, Moreira GMSG, Bertoglio F, Russo G, Hust M. Developing Recombinant Antibodies by Phage Display Against Infectious Diseases and Toxins for Diagnostics and Therapy.. Front Cell Infect Microbiol 2021;11:697876.
- Lai JY, Lim TS. Infectious disease antibodies for biomedical applications: A mini review of immune antibody phage library repertoire.. Int J Biol Macromol 2020 Nov 15;163:640-648.
- Bashir S, Paeshuyse J. Construction of Antibody Phage Libraries and Their Application in Veterinary Immunovirology.. Antibodies (Basel) 2020 Jun 3;9(2).
- Kuhn P, Fühner V, Unkauf T, Moreira GM, Frenzel A, Miethe S, Hust M. Recombinant antibodies for diagnostics and therapy against pathogens and toxins generated by phage display.. Proteomics Clin Appl 2016 Oct;10(9-10):922-948.
- Hülseweh B, Rülker T, Pelat T, Langermann C, Frenzel A, Schirrmann T, Dübel S, Thullier P, Hust M. Human-like antibodies neutralizing Western equine encephalitis virus.. MAbs 2014 May-Jun;6(3):718-27.
- Rülker T, Voß L, Thullier P, O' Brien LM, Pelat T, Perkins SD, Langermann C, Schirrmann T, Dübel S, Marschall HJ, Hust M, Hülseweh B. Isolation and characterisation of a human-like antibody fragment (scFv) that inactivates VEEV in vitro and in vivo.. PLoS One 2012;7(5):e37242.
- Kirsch MI, Hülseweh B, Nacke C, Rülker T, Schirrmann T, Marschall HJ, Hust M, Dübel S. Development of human antibody fragments using antibody phage display for the detection and diagnosis of Venezuelan equine encephalitis virus (VEEV).. BMC Biotechnol 2008 Sep 2;8:66.
- Berry JD. Rational monoclonal antibody development to emerging pathogens, biothreat agents and agents of foreign animal disease: The antigen scale.. Vet J 2005 Sep;170(2):193-211.
- Wang HW, Cole D, Jiang WZ, Jin HT, Fu N, Chen ZL, Jin NY. Engineering and functional evaluation of a single-chain antibody against HIV-1 external glycoprotein gp120.. Clin Exp Immunol 2005 Jul;141(1):72-80.
Use Nutrition Calculator
Check if your horse's diet meets their nutrition requirements with our easy-to-use tool Check your horse's diet with our easy-to-use tool
Talk to a Nutritionist
Discuss your horse's feeding plan with our experts over a free phone consultation Discuss your horse's diet over a phone consultation
Submit Diet Evaluation
Get a customized feeding plan for your horse formulated by our equine nutritionists Get a custom feeding plan formulated by our nutritionists
