Purification of equine IgG3 by lectin affinity and an interaction analysis via microscale thermophoresis

De-Simone, Salvatore Giovanni; Nascimento, Hilton Jorge; Prado, Isis C.; Aguiar, Aniesse Silva; Melgarejo, Anibal R.; Pina, Jorge L.S.; Ferreira, Patricia F.; Provance, David William

doi:10.1016/j.ab.2018.09.010

Cited by 8 publications

(5 citation statements)

References 44 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…To remove BSA-reactive antibodies, the rabbit sera anti-EP8 (rab-EP8) was passed over a Sepharose-4B column (3 cm × 1 cm i.d.) that was coupled with BSA according to conditions previously described [ 37 ]. The removal of anti-BSA antibodies was evaluated by the loss of reactivity in the rab-EP8 to BSA by immunoblot ( Figure S1 ).…”

Section: Methodsmentioning

confidence: 99%

Trypanosoma cruzi Presenilin-Like Transmembrane Aspartyl Protease: Characterization and Cellular Localization

et al. 2020

Self Cite

View full text Add to dashboard Cite

The increasing detection of infections of Trypanosoma cruzi, the etiological agent of Chagas disease, in non-endemic regions beyond Latin America has risen to be a major public health issue. With an impact in the millions of people, current treatments rely on antiquated drugs that produce severe side effects and are considered nearly ineffective for the chronic phase. The minimal progress in the development of new drugs highlights the need for advances in basic research on crucial biochemical pathways in T. cruzi to identify new targets. Here, we report on the T. cruzi presenilin-like transmembrane aspartyl enzyme, a protease of the aspartic class in a unique phylogenetic subgroup with T. vivax separate from protozoans. Computational analyses suggest it contains nine transmembrane domains and an active site with the characteristic PALP motif of the A22 family. Multiple linear B-cell epitopes were identified by SPOT-synthesis analysis with Chagasic patient sera. Two were chosen to generate rabbit antisera, whose signal was primarily localized to the flagellar pocket, intracellular vesicles, and endoplasmic reticulum in parasites by whole-cell immunofluorescence. The results suggest that the parasitic presenilin-like enzyme could have a role in the secretory pathway and serve as a target for the generation of new therapeutics specific to the T. cruzi.

show abstract

Section: Methodsmentioning

confidence: 99%

Trypanosoma cruzi Presenilin-Like Transmembrane Aspartyl Protease: Characterization and Cellular Localization

et al. 2020

Self Cite

View full text Add to dashboard Cite

show abstract

“…Although donkeys are prevalent in China, especially in the rural areas of Northern China, very few therapeutic antibodies were prepared from the donkeys. This may be related to the fact that the use of horse-derived antibodies for therapeutic purposes has a long history, and their preparation methods are relatively mature [ 31 ]. To explore alternative sources of therapeutic antibodies that can mitigate the shortage of anti-CDV antibody, in this study we used donkeys to prepare therapeutic anti-CDV antibody, and analyze its therapeutic effects and safety.…”

Section: Introductionmentioning

confidence: 99%

Donkey-derived anti-CDV IgG, as a passive immunotherapy agent, can effectively increase survival rates of the experimental CDV-infected dogs

et al. 2021

View full text Add to dashboard Cite

Background Humoral immunity plays an important role in the prevention of canine distemper. Anti-CD virus (CDV) antibody has strong antiviral activity and is widely used in the treatment of CD. However, with the increase of CD cases, the availability of therapeutic CD antibody fell short of the clinical needs. Results The high-titer antiserum with the high-titer neutralizing activity against CDV was obtained from the donkeys (Dezhou Donkey) immunized with the inactivated CDV vaccine. The donkey anti-CDV IgG was purified from the donkey serum, which was identified to significantly inhibit the CDV replication in the cultured Vero cells and effectively reduce the clinical symptoms and increase the survival rates (75%) of CDV-infected dogs (Shih-tzu Dog), similar to that treated with the dog-derived anti-CDV IgG. These results indicate that donkey-derived IgG is a potential substitute for dog-derived IgG to treat the CD in clinic. Conclusions Administration of donkey-derived anti-CDV IgG can ameliorate clinical symptoms and inhibit virus replication, thereby increasing the survival of CDV-infected dogs. This study opens up a new source of therapeutic antibody for CD treatment.

show abstract

“…The assay was performed in PBS containing 0.05% Tween 20, and after a short incubation, the samples were loaded into standard MST NT.115 glass capillaries. Kd data were calculated using the NanoTemper software package (MO.Affinity Analysis v2.3) [27].…”

Section: Microscale Thermophoresis (Mst)mentioning

confidence: 99%

Humoral Immune Response to SARS-CoV-2 Spike Protein Receptor-Binding Motif Linear Epitopes

Monteiro,

Lechuga,

Napoleão-Pêgo

et al. 2024

Vaccines

View full text Add to dashboard Cite

The worldwide spread of SARS-CoV-2 has led to a significant economic and social burden on a global scale. Even though the pandemic has concluded, apprehension remains regarding the emergence of highly transmissible variants capable of evading immunity induced by either vaccination or prior infection. The success of viral penetration is due to the specific amino acid residues of the receptor-binding motif (RBM) involved in viral attachment. This region interacts with the cellular receptor ACE2, triggering a neutralizing antibody (nAb) response. In this study, we evaluated serum immunogenicity from individuals who received either a single dose or a combination of different vaccines against the original SARS-CoV-2 strain and a mutated linear RBM. Despite a modest antibody response to wild-type SARS-CoV-2 RBM, the Omicron variants exhibit four mutations in the RBM (S477N, T478K, E484A, and F486V) that result in even lower antibody titers. The primary immune responses observed were directed toward IgA and IgG. While nAbs typically target the RBD, our investigation has unveiled reduced seroreactivity within the RBD’s crucial subregion, the RBM. This deficiency may have implications for the generation of protective nAbs. An evaluation of S1WT and S2WT RBM peptides binding to nAbs using microscale thermophoresis revealed a higher affinity (35 nM) for the S2WT sequence (GSTPCNGVEGFNCYF), which includes the FNCY patch. Our findings suggest that the linear RBM of SARS-CoV-2 is not an immunodominant region in vaccinated individuals. Comprehending the intricate dynamics of the humoral response, its interplay with viral evolution, and host genetics is crucial for formulating effective vaccination strategies, targeting not only SARS-CoV-2 but also anticipating potential future coronaviruses.

show abstract

Purification of equine IgG3 by lectin affinity and an interaction analysis via microscale thermophoresis

Cited by 8 publications

References 44 publications

Trypanosoma cruzi Presenilin-Like Transmembrane Aspartyl Protease: Characterization and Cellular Localization

Trypanosoma cruzi Presenilin-Like Transmembrane Aspartyl Protease: Characterization and Cellular Localization

Donkey-derived anti-CDV IgG, as a passive immunotherapy agent, can effectively increase survival rates of the experimental CDV-infected dogs

Humoral Immune Response to SARS-CoV-2 Spike Protein Receptor-Binding Motif Linear Epitopes

Contact Info

Product

Resources

About