Biological Activity of the Carrier as a Factor in Immunogen Design for Haptens

Ertekin, Özlem; Akçael, Esin; Kocaağa, Harun; Öztürk, Şule

doi:10.3390/molecules23112977

Cited by 14 publications

(7 citation statements)

References 40 publications

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“…We speculated that failure to increase the titer of Ent-specific IgY in egg yolk in Trial#1 could be addressed by optimizing immunization conditions, such as the choice of the carrier protein ( 33 ), layer breed ( 34 ), and immunization route ( 35 ). Therefore, to test this, in pilot Trial #2, three different carrier proteins (BSA, KLH and recombinant CmeC) were used to prepare Ent conjugates.…”

Section: Resultsmentioning

confidence: 99%

Evaluation of the Immunogenic Response of a Novel Enterobactin Conjugate Vaccine in Chickens for the Production of Enterobactin-Specific Egg Yolk Antibodies

et al. 2021

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Passive immunization with specific egg yolk antibodies (immunoglobulin Y, IgY) is emerging as a promising alternative to antibiotics to control bacterial infections. Recently, we developed a novel conjugate vaccine that could trigger a strong immune response in rabbits directed against enterobactin (Ent), a highly conserved siderophore molecule utilized by different Gram-negative pathogens. However, induction of Ent-specific antibodies appeared to be affected by the choice of animal host and vaccination regimen. It is still unknown if the Ent conjugate vaccine can trigger a specific immune response in layers for the purpose of production of anti-Ent egg yolk IgY. In this study, three chicken vaccination trials with different regimens were performed to determine conditions for efficient production of anti-Ent egg yolk IgY. Purified Ent was conjugated to three carrier proteins, keyhole limpet hemocyanin (KLH), bovine serum albumin (BSA) and CmeC (a subunit vaccine candidate), respectively. Intramuscular immunization of Barred Rock layers with KLH-Ent conjugate four times induced strong immune response against whole conjugate vaccine but the titer of Ent-specific IgY did not change in yolk with only a 4 fold increase detected in serum. In the second trial, three different Ent conjugate vaccines were evaluated in Rhode Island Red pullets with four subcutaneous injections. The KLH-Ent or CmeC-Ent conjugate consistently induced high level of Ent-specific IgY in both serum (up to 2,048 fold) and yolk (up to 1,024 fold) in each individual chicken. However, the Ent-specific immune response was only temporarily and moderately induced using a BSA-Ent vaccination. In the third trial, ten White Leghorn layers were subcutaneously immunized three times with KLH-Ent, leading to consistent and strong immune response against both whole conjugate and the Ent molecule in each chicken; the mean titer of Ent-specific IgY increased approximately 32 and 256 fold in serum and yolk, respectively. Consistent with its potent binding to various Ent derivatives, the Ent-specific egg yolk IgY also inhibited in vitro growth of a representative Escherichia coli strain. Together, this study demonstrated that the novel Ent conjugate vaccine could induce strong, specific, and robust immune response in chickens. The Ent-specific hyperimmune egg yolk IgY has potential for passive immune intervention against Gram-negative infections.

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Section: Resultsmentioning

confidence: 99%

Evaluation of the Immunogenic Response of a Novel Enterobactin Conjugate Vaccine in Chickens for the Production of Enterobactin-Specific Egg Yolk Antibodies

et al. 2021

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“…The solution to this is usually in the conjugation of small antigens with very immunogenic, large carrier proteins. [45] However, in general, it induces a strong immune response specific for carrier proteins and not against the conjugated peptide. Therefore, to create the best immunogen, a recommendation might be to prepare the conjugates with several different carriers and a range of pepide-carrier, protein-coupling ratios, which takes time and resources.…”

Section: Cbs2a21p75 Mab Affinity Constant Determinationmentioning

confidence: 99%

Untitled

2023

BPJ

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T he SARS-CoV-2 spike protein S2 subunit plays an essential role in the virus-host cell membrane fusion process. Therefore, the subject of this study was to characterize the gamma-immunoglobulin (IgG) response, in a group of COVID-19 convalescent patients, against the S2 subunit with eight linear peptides to generate a monoclonal antibody (mAb) against the immunodominant linear peptide to be used for therapeutic and diagnostic purposes. Results of antibody percentages against assessed linear peptides were 100% for A21P73, A21P74, A21P75, A21P76, M20P51, M20P65, M20P83, and 66.7% for M20P85. Plasma samples were also used for purifying IgG to corroborate specificity against the same linear peptides, where results reproduced those applying plasmas directly to ELISA-plates. Within these peptides, A21P75 was chosen as immunodominant (100% of recognition with higher absorbance). The A21P75 linear peptide showed poor immunogenicity in mice (1:4000-8000 after four doses), allowing the generation of a CB.HS2A21P75 hybridoma for mAb production that recognized the A21P75 linear peptide with middle-to-high affinity constant (K aff ) (0.8×10 8 M -1 ).This study concludes that the A21P75 linear peptide is the assessed immunodominant linear peptide for this COVID-19 convalescent patient group. This peptide is located in the HR1 region that plays an important role in the SARS-CoV-2 host cell membrane fusion process and is highly conserved between SARS-CoV-2 and SARS-CoV. Thus, due to CB.S2A21P75 mAb specificity and K aff , it might be the proper reagent to study inhibition of virushost cell membrane fusion, and as a diagnostic reagent for coronavirus. Finally, the combination of A21P75 linear peptide with other peptides (e.g., receptor binding domain [RBD]) could be suitable reagents for the development of vaccines and therapeutic antibodies with virus infection-blocking capacity.

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“…The covalent binding of antibodies, oligonucleotide chains, or phages was achieved by modifying active groups such as −NH, −OH, −COOH, or other functional groups on the surface of magnetic cores. 15,16 The magnetic nanoparticles (MNPs) with specific ligands formed an "immunomagnetic microsphere-target" complex with the target substance through the antigen−antibody specific reaction 17 and moved directionally under the action of a specific magnetic field to achieve the separation of the target substance from the complex. 18−20 In recent years, functionalized core−shell magnetic nanoparticles (FMNPs) have developed rapidly in the fields of immunoassay, cell separation, protein purification, etc.…”

Section: Introductionmentioning

confidence: 99%

Efficient Hapten-Specific Biopanning Strategy Based on the Fe₃O₄@ENR-Functionalized Core–Shell Magnetic Nanoparticles

Liu,

Chen

et al. 2023

Langmuir

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The biopanning of target-specific phages is one of the most critical steps in the preparation of single-domain antibodies. In the traditional biopanning of haptens, the nonspecific binding of library phages to macromolecular proteins is one of the most challenging problems in preparing single-domain antibodies. In this research, Fe3O4@ENR-functionalized core–shell magnetic nanoparticles (FMNPs) were silylated and aminated by tetraethyl orthosilicate and (3-aminopropyl)triethoxysilane, and target enrofloxacin was coupled onto the surface by the carbodiimide method. The magnetic nanoparticles were characterized by Fourier transform infrared spectroscopy, particle size distribution, zeta potential, transmission electron microscopy observation, and indirect enzyme-linked immunosorbent assay (ELISA). A biopanning strategy based on Fe3O4@ENR FMNPs was then established to solve the problem in the traditional solid-phase biopanning process. The results showed that a considerable number of enrofloxacin (ENR)-positive phages were screened by only one round of biopanning. Finally, two ENR-specific shark-derived single-domain genes were identified and validated by monoclonal phage ELISA, gene sequencing, and biolayer interferometry technology. Our study provides a new biopanning strategy based on Fe3O4@ENR FMNPs for efficiently providing phages specific to haptens.

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Biological Activity of the Carrier as a Factor in Immunogen Design for Haptens

Cited by 14 publications

References 40 publications

Evaluation of the Immunogenic Response of a Novel Enterobactin Conjugate Vaccine in Chickens for the Production of Enterobactin-Specific Egg Yolk Antibodies

Evaluation of the Immunogenic Response of a Novel Enterobactin Conjugate Vaccine in Chickens for the Production of Enterobactin-Specific Egg Yolk Antibodies

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Efficient Hapten-Specific Biopanning Strategy Based on the Fe₃O₄@ENR-Functionalized Core–Shell Magnetic Nanoparticles

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Biological Activity of the Carrier as a Factor in Immunogen Design for Haptens

Cited by 14 publications

References 40 publications

Evaluation of the Immunogenic Response of a Novel Enterobactin Conjugate Vaccine in Chickens for the Production of Enterobactin-Specific Egg Yolk Antibodies

Evaluation of the Immunogenic Response of a Novel Enterobactin Conjugate Vaccine in Chickens for the Production of Enterobactin-Specific Egg Yolk Antibodies

Untitled

Efficient Hapten-Specific Biopanning Strategy Based on the Fe3O4@ENR-Functionalized Core–Shell Magnetic Nanoparticles

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Efficient Hapten-Specific Biopanning Strategy Based on the Fe₃O₄@ENR-Functionalized Core–Shell Magnetic Nanoparticles