Identification of structural variations in the carboxyl terminus of Alzheimer's disease-associated <i>β</i>A4[1–42] amyloid using a monoclonal antibody

Jayasena, U. L. H. R.; Gribble, S; McKenzie, A L; Beyreuther, Konrad; Masters, Colin L.; Underwood, John R.

doi:10.1046/j.1365-2249.2001.01209.x

Cited by 3 publications

(12 citation statements)

References 19 publications

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“…Fab antibody gene fragments from the mouse monoclonal hybridoma cell line 1E8 [17–22] were generated by PCR using primers generated from immunological sequences in Kabat et al. [20].…”

Section: Resultsmentioning

confidence: 99%

“…Epitope mapping (Fig. 7) showed that the 1E8–4b recombinant fragment and the 1E8 antibody recognized the following Aβ peptides: Aβ[5–19], Aβ[8–22], Aβ[11–25], Aβ[14–28] and Aβ[17–31], with 1E8 exhibiting much stronger binding than 1E8–4b against theAβ[5–19] peptide.…”

Section: Resultsmentioning

confidence: 99%

“…1E8–4b showed a weaker response to Aβ[17–19] than 1E8 due perhaps to altered conformation or unique requirements of only Aβ18,19/19] plus [20–22]. Overall, 1E8 and 1E8–4b contained a specific epitope encompassing Aβ[17,18,18–22].…”

Section: Discussionmentioning

confidence: 96%

“…To determine the specificity of the Aβ parent and Fab antibodies, Pepset analysis was undertaken comprising a set of 15 mer overlapping biotinylated peptides incrementing by three amino acids per peptide and encompassing the human [695–652] sequence (Chiron Mimotopes Pty Ltd, Melbourne, Australia) [18]. The 96‐well microtitre Cliniplates EB (Labsystems, Helsinki, Finland) were incubated with 500 ng/well streptavidin at 37°C overnight and evaporated to dryness.…”

Section: Methodsmentioning

confidence: 99%

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Generation of a recombinant Fab antibody reactive with the Alzheimer's disease-related Aβ peptide

Tammer

Coia

Cappai

et al. 2002

Clinical and Experimental Immunology

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SUMMARYA recombinant Fab antibody, designated 1E8-4b, which reacts with the Alzheimer's disease (AD)-related Ab peptides, Ab , Ab and Ab has been developed. The 1E8-4b Fab was constructed by cloning the V H C H1 and V L C L domains from the parent hybridoma 1E8 antibody, reported previously to recognize these Ab peptides. Briefly, a C-terminal Flag tag sequence was incorporated into this construct, which was ligated into the vector pHFA 2 and expressed in Escherichia coli. Following purification on an M2 anti-Flag affinity column, the 1E8-4b recombinant Fab antibody was shown to bind plaques within sections of brain tissue from CERAD-defined AD patients by immunohistochemistry. ELISA, epitope mapping and immunoblotting confirmed the recognition of the Ab[1-40/42/43] peptides by the 1E8-4b Fab. The 1E8-4b Fab did not recognize APP695 or APP770 which contain the Ab sequence. The Ab specificity of the recombinant 1E8-4b Fab antibody was identical to the parent 1E8 monoclonal antibody.

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“…Fab antibody gene fragments from the mouse monoclonal hybridoma cell line 1E8 [17–22] were generated by PCR using primers generated from immunological sequences in Kabat et al. [20].…”

Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 96%

Section: Methodsmentioning

confidence: 99%

See 2 more Smart Citations

Generation of a recombinant Fab antibody reactive with the Alzheimer's disease-related Aβ peptide

Tammer

Coia

Cappai

et al. 2002

Clinical and Experimental Immunology

Self Cite

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“…Remodeling protein conformation therefore exposes different structural determinants that can be recognized by different antibodies. Such antibodies have been used to discriminate between the inactive and active conformations of a protein [72], to neutralize viruses [73] and to differentiate between the aggregated or monomeric form of proteins [74][75][76]. When proteins are adsorbed on material surfaces, some epitopes are masked, which reduces the binding of the corresponding antibodies [77][78][79].…”

Section: Evidences Of Material-induced Protein Conformational Changesmentioning

confidence: 99%

Protein conformational changes induced by adsorption onto material surfaces: an important issue for biomedical applications of material science

Ballet¹,

Boulangé²,

Bréchet³

et al. 2010

Bulletin of the Polish Academy of Sciences: Technical Sciences

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Abstract. Protein adsorption on solid surfaces is a widespread phenomenon of large biological and biotechnological significance. Conformational changes are likely to accompany protein adsorption, but are difficult to evidence directly. Nevertheless they have important consequences, since the partial unfolding of protein domains can expose hitherto hidden amino acids. This remodeling of the protein surface can trigger the activation of molecular complexes such as the blood coagulation cascade or the innate immune complement system. In the case of extracellular matrix, it can also change the way cells interact with the material surfaces and result in modified cell behavior. In this review, we present direct and indirect evidences that support the view that some proteins change their conformation upon adsorption. We also show that both physical and chemical methods are needed to study the extent and kinetics of protein conformational changes. In particular, AFM techniques and cryo-electron microscopy provide useful and complementary information. We then review the chemical and topological features of both proteins and material surfaces in relation with protein adsorption. Mutating key amino acids in proteins changes their stability and this is related to material-induced conformational changes, as shown for instance with insulin. In addition, combinatorial methods should provide valuable information about peptide or antibody adsorption on well-defined material surfaces. These techniques could be combined with molecular modeling methods to decipher the rules governing conformational changes associated with protein adsorption.

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Based on mRNA Sequencing Techniques to Explore the Molecular Mechanism of Buzhong Yiqi Decoction for Autoimmune Thyroiditis

Liu

Song

et al. 2024

CCHTS

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Objective: Autoimmune diseases (AD) account for a high percentage of the population. One of the most prevalent is autoimmune thyroiditis (AIT). However, the therapeutic effects of Buzhong Yiqi (BZYQ) decoction on AIT have not been studied yet. The majority of the present study was conducted on NOD.H-2h4 mice in an attempt to ascertain the therapeutic effects of BZYQ decoction on AIT. Methods: The 0.05% sodium iodide water (NaI)-induced AIT mice model was established. A total of nine NOD.H-2h4 mice were randomly divided into three groups: the normal group provided with regular water, the model group drinking freely 0.05% NaI, and the treatment group treated with BZYQ decoction (9.56 g/kg) after NaI supplementation (NaI + BZYQ). BZYQ decoction was administered orally once daily for eight weeks. The thyroid histopathology test was used to measure the severity of lymphocytic infiltration. An enzyme-linked immunosorbent assay (ELISA) was used to determine the levels of anti-thyroglobulin antibody (TgAb), interleukin (IL)-1β, IL-6, and IL-17. The Illumina HiSeq X sequencing platform was utilized to analyze the thyroid tissue by mRNA expression profiles. Bioinformatics analysis was used to investigate the biological function of the differentially expressed mRNAs. In addition, the expression of Carbonyl Reductase 1 (CBR1), 6-Pyruvoyltetrahydropterin Synthase (PTS), Major Histocompatibility Complex, Class II (H2-EB1), Interleukin 23 Subunit Alpha (IL-23A), Interleukin 6 Receptor (IL-6RA), and Janus Kinase 1 (JAK1) was measured by quantitative real-time PCR (qRT-PCR). Results: The treatment group exhibited significantly lower rates of thyroiditis and lymphocyte infiltration compared to the model group. Serum levels of TgAb, IL-1β, IL-6, and IL-17 were significantly higher in the model group, but they fell dramatically after BZYQ decoction administration. According to our results, 495 genes showed differential expression in the model group compared to the control group. Six hundred twenty-five genes were significantly deregulated in the treatment group compared to the model group. Bioinformatic analysis showed that most mRNAs were associated with immune-inflammatory responses and were involved in multiple signaling pathways, including folate biosynthesis and the Th17 cell differentiation pathway. CBR1, PTS, H2-EB1, IL23A, IL-6RA and JAK1 mRNA participated in folate biosynthesis and the Th17 cell differentiation pathway. The qRT-PCR analysis confirmed that the above mRNAs were regulated in the model group compared to the treatment group. Conclusion: The results of this investigation have revealed novel insights into the molecular mechanism of action of BZYQ decoction against AIT. The mechanism may be partially attributed to the regulation of mRNA expression and pathways.

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Identification of structural variations in the carboxyl terminus of Alzheimer's disease-associated βA4[1–42] amyloid using a monoclonal antibody

Cited by 3 publications

References 19 publications

Generation of a recombinant Fab antibody reactive with the Alzheimer's disease-related Aβ peptide

Generation of a recombinant Fab antibody reactive with the Alzheimer's disease-related Aβ peptide

Protein conformational changes induced by adsorption onto material surfaces: an important issue for biomedical applications of material science

Based on mRNA Sequencing Techniques to Explore the Molecular Mechanism of Buzhong Yiqi Decoction for Autoimmune Thyroiditis

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