Development of glutamic acid decarboxylase 65 (GAD65) autoantibody assay using biotin–GAD65 fusion protein

Luo, Dong; Rogers, Chad N.; Steed, Jordan; Gilliam, Lisa K.; Hampe, Christiane S.; Lernmark, Åke

doi:10.1016/j.jbiotec.2004.03.009

Cited by 7 publications

(2 citation statements)

References 27 publications

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“…A fter the development of ELISA in the 70s (1,2), the discovery of the high avidin-biotin affinity (3,4) allowed researchers to precisely manipulate proteins/peptides in numerous ELISA constructs in the 90s (5). Today, although ELISA is used worldwide, for instance to detect and quantify Abs, some studies highlighted the influence of experimental parameters on the outputs of such assay (6)(7)(8)(9)(10), reporting that a small change in the structure of an epitope-presenting peptide could impact its recognition by Abs (11)(12)(13)(14) and thus our perception of the analyzed biological processes. This is particularly important for autoimmune diseases such as rheumatoid arthritis (RA) in which autoantibody profiling, specificity, and titers can be associated with pathophysiology mechanisms and impact diseases diagnosis or therapy management (15)(16)(17)(18)(19)(20).…”

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confidence: 99%

Seropositivity and Antibody Profiling of Patients Are Dramatically Impacted by the Features of Peptides Used as Immunosorbents: A Lesson from Anti–Citrullinated Protein/Peptide Antibody

Cornillet

Babos

Magyar

et al. 2018

The Journal of Immunology

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Quantification of Abs toward a single epitope is critical to understanding immunobiological processes. In autoimmunity, the prognostic value of the serological profiles of patients draws much attention, but the detection of Abs toward a single epitope is not well controlled. Particularly, the rheumatoid arthritis (RA)–specific anti–citrullinated protein/peptide Abs (ACPA) are specific to a two-atom change on arginyl residues and are considered a heterogeneous family of Abs. As a model, we studied ACPA to decipher how peptide features used as immunosorbent impact Ab detection. We synthesized 30 peptides encompassing immunodominant epitopes of citrullinated fibrin differing by their length and biotin location and tested them using ELISA with 120 sera from RA and non-RA rheumatic disease controls, generating over 3000 experimental measurements. We showed that minor molecular changes in peptide chemical structure had dramatic consequences. Even when peptides exhibited the same epitope, measured Ab titers were extremely variable, and patients’ seropositivity was discordant in up to 50% of cases. The distance between epitope and biotin was the most critical parameter for efficient Ab detection irrespective of biotin position or peptide length. Finally, we identified a 15-mer peptide bearing a single citrullinated epitope detecting almost all ACPA-positive sera, thus revealing a high degree of homogeneity in RA autoimmune response. This integrative analysis deciphers the dramatic impact of the molecular design of peptide-based technologies for epitope-specific Ab quantification. It provides a model for assay development and highlights that the studies using such technologies can give a wrong perception of biological processes and therefore that medical use of data must be cautious.

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mentioning

confidence: 99%

Seropositivity and Antibody Profiling of Patients Are Dramatically Impacted by the Features of Peptides Used as Immunosorbents: A Lesson from Anti–Citrullinated Protein/Peptide Antibody

Cornillet

Babos

Magyar

et al. 2018

The Journal of Immunology

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“…2 shows the correlation between the TAXI levels obtained on two different days. These CV values are in the range of values described for methods to quantify other compounds in plant and animal tissues (Silvan et al 1993;Assis et al 2002;van den Heuvel et al 2003;Luo et al 2004).…”

Section: Repeatability Of Quantification Techniquesmentioning

confidence: 76%

Quantification of Wheat TAXI and XIP Type Xylanase Inhibitors: A Comparison of Analytical Techniques

Gebruers

Beaugrand²,

Croes

et al. 2008

Cereal Chem

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To date, three different techniques are available for the quantification of TAXI and XIP type proteinaceous xylanase inhibitors in cereals. A first approach is based on the determination of the residual activities of xylanases (also referred to as endo‐1,4‐β‐d‐xylanases, EC 3.2.1.8), which are specifically inhibited by these inhibitors, after incubation with sample containing the inhibitors. The other two techniques are immunoblotting and ELISA which are based on recognition of TAXI and XIP proteins by specific antibodies. TAXI, as well as XIP, are easily extracted by aqueous buffers. Hence, the large difference in their concentrations (2–10 fold higher for XIP than for TAXI in whole meal) is not caused by differences in extractability. The repeatabilities of the three techniques are comparable. The intra‐assay and inter‐assay coefficients of variation were 6–7 and 10–14%, respectively, which is in the range of values described for methods to quantify other compounds in plant and animal tissues. The three methods give comparable results, suggesting they have similar accuracies. The choice of the technique to be used will depend not only on the sensitivity and dynamic range needed, but also on its technical simplicity and the need for high‐throughput analysis.

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Development of 2 alternative enzyme-linked immunosorbent assays for routine screening of glutamic acid decarboxylase autoantibodies

Villalba

Valdez

Iacono

et al. 2007

Clinica Chimica Acta

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Development of glutamic acid decarboxylase 65 (GAD65) autoantibody assay using biotin–GAD65 fusion protein

Cited by 7 publications

References 27 publications

Seropositivity and Antibody Profiling of Patients Are Dramatically Impacted by the Features of Peptides Used as Immunosorbents: A Lesson from Anti–Citrullinated Protein/Peptide Antibody

Seropositivity and Antibody Profiling of Patients Are Dramatically Impacted by the Features of Peptides Used as Immunosorbents: A Lesson from Anti–Citrullinated Protein/Peptide Antibody

Quantification of Wheat TAXI and XIP Type Xylanase Inhibitors: A Comparison of Analytical Techniques

Development of 2 alternative enzyme-linked immunosorbent assays for routine screening of glutamic acid decarboxylase autoantibodies

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