TY - JOUR
A1 - Deutschmann, Claudia
A1 - Roggenbuck, Dirk
A1 - Schierack, Peter
A1 - Rödiger, Stefan
T1 - Autoantibody testing by enzyme-linked immunosorbent assay-a case in which the solid phase decides on success and failure
JF - Heliyon
N2 - Background: The enzyme-linked immunosorbent assay (ELISA) is an indispensable tool for clinical diagnostics to identify or differentiate diseases such as autoimmune illnesses, but also to monitor their progression or control the efficacy of drugs. One use case of ELISA is to differentiate between different states (e.g. healthy vs. diseased). Another goal is to quantitatively assess the biomarker in question, like autoantibodies. Thus, the ELISA technology is used for the discovery and verification of new autoantibodies, too. Of key interest, however, is the development of immunoassays for the sensitive and specific detection of such biomarkers at early disease stages. Therefore, users have to deal with many parameters, such as buffer systems or antigen-autoantibody interactions, to successfully establish an ELISA. Often, fine-tuning like testing of several blocking substances is performed to yield high signal-to-noise ratios.
Methods: We developed an ELISA to detect IgA and IgG autoantibodies against chitinase-3-like protein 1 (CHI3L1), a newly identified autoantigen in inflammatory bowel disease (IBD), in the serum of control and disease groups (n = 23, respectively). Microwell plates with different surface modifications (PolySorp and MaxiSorp coating) were tested to detect reproducibility problems.
Results: We found a significant impact of the surface properties of the microwell plates. IgA antibody reactivity was significantly lower, since it was in the range of background noise, when measured on MaxiSorp coated plates (p < 0.0001). The IgG antibody reactivity did not differ on the diverse plates, but the plate surface had a significant influence on the test result (p = 0.0005).
Conclusion: With this report, we want to draw readers' attention to the properties of solid phases and their effects on the detection of autoantibodies by ELISA. We want to sensitize the reader to the fact that the choice of the wrong plate can lead to a false negative test result, which in turn has serious consequences for the discovery of autoantibodies.
KW - biochemistry
KW - coatings
KW - surface chemistry
KW - immunology
KW - proteins
KW - laboratory medicine
KW - clinical research
KW - enzyme-linked immunosorbent
KW - assay
KW - biomarker discovery
KW - reproducibility
KW - solid-phase
KW - autoantibody
Y1 - 2020
U6 - https://doi.org/10.1016/j.heliyon.2020.e03270
SN - 2405-8440
VL - 6
IS - 1
PB - Elsevier
CY - London [u.a.]
ER -
TY - JOUR
A1 - Schönemann, Eric
A1 - Koc, Julian
A1 - Aldred, Nick
A1 - Clare, Anthony S.
A1 - Laschewsky, André
A1 - Rosenhahn, Axel
A1 - Wischerhoff, Erik
T1 - Synthesis of Novel Sulfobetaine Polymers with Differing Dipole Orientations in Their Side Chains, and Their Effects on the Antifouling Properties
JF - Macromolecular rapid communications
N2 - The impact of the orientation of zwitterionic groups, with respect to the polymer backbone, on the antifouling performance of thin hydrogel films made of polyzwitterions is explored. In an extension of the recent discussion about differences in the behavior of polymeric phosphatidylcholines and choline phosphates, a quasi-isomeric set of three poly(sulfobetaine methacrylate)s is designed for this purpose. The design is based on the established monomer 3-[N-2-(methacryloyloxy)ethyl-N,N-dimethyl]ammonio-propane-1-sulfonate and two novel sulfobetaine methacrylates, in which the positions of the cationic and the ionic groups relative to the polymerizable group, and thus also to the polymer backbone, are altered. The effect of the varied segmental dipole orientation on their water solubility, wetting behavior by water, and fouling resistance is compared. As model systems, the adsorption of the model proteins bovine serum albumin (BSA), fibrinogen, and lysozyme onto films of the various polyzwitterion surfaces is studied, as well as the settlement of a diatom (Navicula perminuta) and barnacle cyprids (Balanus improvisus) as representatives of typical marine fouling communities. The results demonstrate the important role of the zwitterionic group's orientation on the polymer behavior and fouling resistance.
KW - antifouling
KW - coatings
KW - crosslinking
KW - hydrophilic polymers
KW - polyzwitterions
Y1 - 2019
U6 - https://doi.org/10.1002/marc.201900447
SN - 1022-1336
SN - 1521-3927
VL - 41
IS - 1
PB - Wiley-VCH
CY - Weinheim
ER -