Serogroup-reactive and type-specific detection of bluetongue virus antibodies using chicken scFvs in inhibition ELISAs

Fehrsen, Jeanni; Wyngaardt, Wouter Van; Mashau, Cordelia; Potgieter, A. C.; Chaudhary, Vijay; Gupta, Amita; Jordaan, Frances; Plessis, D.H. Du

doi:10.1016/j.jviromet.2005.04.015

Cited by 23 publications

(23 citation statements)

References 31 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Among various antibody forms, the antigen-binding fragment or singlechain variable fragment (scFv) displayed on the phage is highly suitable and fast for selecting specific antibodies (23)(24)(25). Among the many animals that are adequate as animal models for producing antibodies against immunizing antigens, chickens are the most convenient and rapid host for constructing antibody libraries for selecting specific scFvs against various targets for therapy or diagnosis (24,26,27). Monoclonal scFv is a small protein with favorable tissue penetration, maintaining the variable regions of light and heavy chains that are joined with a flexible peptide linker, and it has a specific antigen-binding ability (28,29).…”

mentioning

confidence: 99%

Antibodies against Venom of the Snake Deinagkistrodon acutus

Lee

Liang

et al. 2016

Appl Environ Microbiol

View full text Add to dashboard Cite

h Snake venom protein from Deinagkistrodon acutus (DA protein), one of the major venomous species in Taiwan, causes hemorrhagic symptoms that can lead to death. Although horse-derived antivenin is a major treatment, relatively strong and detrimental side effects are seen occasionally. In our study, yolk immunoglobulin (IgY) was purified from eggs, and DA protein was recognized using Western blotting and an enzyme-linked immunosorbent assay (ELISA), similar to therapeutic horse antivenin. The ELISA also indicated that specific IgY antibodies were elicited after the fifth booster, plateaued, and lasted for at least 3 months. To generate monoclonal single-chain variable fragment (scFv) antibodies, we used phage display technology to construct two libraries with short or long linkers, containing 6.24 ؋ 10 8 and 5.28 ؋ 10 8 transformants, respectively. After four rounds of biopanning, the eluted phage titer increased, and the phage-based ELISA indicated that the specific clones were enriched. Nucleotide sequences of 30 individual clones expressing scFv were analyzed and classified into four groups that all specifically recognized the DA venom protein. Furthermore, based on mass spectrometry, the scFv-bound protein was deduced to be snake venom metalloproteinase proteins. Most importantly, both IgY and mixed scFv inhibited the lethal effect in mice injected with the minimum lethal dosage of the DA protein. We suggest that together, these antibodies could be applied to the development of diagnostic agents or treatments for snakebite envenomation in the future.

show abstract

mentioning

confidence: 99%

Antibodies against Venom of the Snake Deinagkistrodon acutus

Lee

Liang

et al. 2016

Appl Environ Microbiol

View full text Add to dashboard Cite

show abstract

“…Moreover, many conserved proteins that are not immunogenic in mammals yield high-titer antibodies when used to immunize chickens (8,10). Chicken scFvs have been previously generated against bluetongue virus, the SARS-CoV spike protein, prion proteins, and infectious bursal disease virus (7,17,23,29).…”

Section: Introductionmentioning

confidence: 99%

Neutralization Analysis of a Chicken Single-Chain Variable Fragment Derived from an Immune Antibody Library Against Infectious Bronchitis Virus

Lin

et al. 2015

Viral Immunology

View full text Add to dashboard Cite

Avian infectious bronchitis virus (IBV), which is prevalent in many countries causing severe economic loss to the poultry industry, causes infectious bronchitis (IB) in birds. Recombinant single-chain variable fragments (scFvs) have been proven to effectively inhibit many viruses, both in vitro and in vivo, and they could be a potential diagnostic and therapeutic reagent to control IB. In this study, six anti-IBV chicken scFvs, ZL.10, ZL.64, ZL.78, ZL.80, ZL.138, and ZL.256, were obtained by screening random clones from an immune antibody library. An analysis of nucleotide sequences revealed that they represented distinctive genetic sequences and greatly varied in complementarity-determining region three of the heavy chain. Neutralization tests showed that ZL.10, which bound the S1 protein in western blots, inhibited the formation of syncytia in Vero cells 48 h post IBV infection and decreased the transcriptional level of nucleoprotein mRNA to 17.2%, while the other five scFvs, including ZL.78 and ZL.256, that bound the N protein did not. In conclusion, the results suggested that specific and neutralizing chicken scFvs against IBV, which can be safe and economical antibody reagents, can be produced in vitro through prokaryotic expression.

show abstract

“…Single-chain variable fragments (scFvs) based on chicken immunoglobulins are particularly suitable as a basis for developing immunotests, not only because they are serologically distinct from mammalian immunoglobulins, but because the avian antibody repertoire can be accessed [9][10][11] more readily than that of any mammal other than the camelids [12]. Recombinant chicken antibodies derived either from large ''universal'' or dedicated immune repertoires have been shown to be eminently useful in a variety of immunotests aimed at detecting either antigens or antibodies [13][14][15][16][17][18][19]. This suggests that if judiciously applied, they could also play a role in the diagnosis of tuberculosis.…”

Section: Introductionmentioning

confidence: 99%

Chicken scFvs and bivalent scFv-CH fusions directed against HSP65 of Mycobacterium bovis

et al. 2010

View full text Add to dashboard Cite

a b s t r a c tTwo chicken single-chain variable region antibody fragments (scFvs) that recognised the 65 kDa heatshock protein (HSP65) of Mycobacterium bovis were selected from a large semi-synthetic phage displayed library. Both recognised HSP65 in indirect enzyme-linked immunosorbent assay (ELISA) and immunoblots and retained their activity during storage. Neither, however, could function as the capture reagent in a sandwich ELISA when immobilised on polystyrene. To establish whether they could be engineered for general use in immunotests, the genes coding for these scFvs were subcloned in expression vectors that contained sequences encoding chicken IgY heavy-chain constant region domains. This resulted in larger bivalent constructs which more closely resembled IgY molecules. The engineered fragments were evaluated in ELISAs and gold-conjugated immunochromatographic tests (ICTs). In contrast to their previous behaviour as scFvs, the modified fragments (designated ''gallibodies'') could be used for immunocapture in ELISA and could be readily conjugated to colloidal gold nanoparticles. A sandwich ICT that could detect recombinant HSP65 was also devised. Although converting the recombinant single-chain monomeric antibody fragments to bivalent immunoglobulin-like molecules did not entirely 'standardise' the behaviour of the scFvs, this approach remains potentially useful for developing practical, robust, immunodiagnostic reagents.

show abstract

Serogroup-reactive and type-specific detection of bluetongue virus antibodies using chicken scFvs in inhibition ELISAs

Cited by 23 publications

References 31 publications

Antibodies against Venom of the Snake Deinagkistrodon acutus

Antibodies against Venom of the Snake Deinagkistrodon acutus

Neutralization Analysis of a Chicken Single-Chain Variable Fragment Derived from an Immune Antibody Library Against Infectious Bronchitis Virus

Chicken scFvs and bivalent scFv-CH fusions directed against HSP65 of Mycobacterium bovis

Contact Info

Product

Resources

About