When developing immunochemical test systems, it is necessary to obtain specific antibodies. Their quality depends, among other things, on the immunogen used. When preparing hapten-protein conjugates to obtain antibodies for low-molecular-weight compounds, the key factors are the structure of the hapten itself, the presence of a spacer, the size of the carrier protein and the degree of its modification by hapten molecules. This work shows that one additional factor-the conditions for obtaining the hapten-protein conjugate-is overlooked. In this work, we have synthesized conjugates of bisphenol A derivative 4,4-bis(hydroxyphenyl)valeric acid (BVA), the protein carrier soybean trypsin inhibitor (STI), and bovine serum albumin (BSA) in reaction media combining water with two organic solvents: dimethylformamide (DMF) or dimethyl sulfoxide (DMSO). Namely, BSA-BVA, STI-BVA, BSA-BVA and STI-BVA conjugates were obtained. Rabbit polyclonal antibodies against the BSA-BVA conjugate demonstrated basically different interactions in the developed ELISA systems using either STI-BVA or STI-BVA conjugates. The use of the STI-BVA conjugate demonstrated the absence of competition in combination with antisera obtained from BSA-BVA in an ELISA. A competitive interaction was observed only with the use of the STI-BVA conjugate. Under the selected conditions, the detection limit of bisphenol A was 8.3 ng/mL, and the working range of determined concentrations was 18.5-290.3 ng/mL. The obtained data demonstrate the possibility of achieving sensitive immunoassays by simply varying the reaction media for the hapten-protein conjugation, which could provide an additional tool in the development of immunoassays for other low-molecular-weight compounds.
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http://dx.doi.org/10.3390/antib13040089 | DOI Listing |
Toxins (Basel)
December 2024
Immunopathology Laboratory, Butantan Institute, São Paulo 05585-090, Brazil.
Jararhagin-C (JarC) is a protein from the venom of consisting of disintegrin-like and cysteine-rich domains. JarC shows a modulating effect on angiogenesis and remodeling of extracellular matrix constituents, improving wound healing in a mouse experimental model. JarC is purified from crude venom, and the yield is less than 1%.
View Article and Find Full Text PDFCurr Issues Mol Biol
December 2024
Faculty of Veterinary Medicine, Ss. Cyril & Methodius University in Skopje, 1000 Skopje, North Macedonia.
The kidney plays an essential role in the proper homeostasis of glucose. In the kidney, glucose transport is carried out across cell membranes by two families of glucose transporters-facilitated diffusion glucose transporters (GLUTs) and Na(+)-dependent glucose co-transporters (SGLT family). Among the transporters, sodium-dependent glucose co-transporters play a major role in the kidney's ability to reabsorb glucose.
View Article and Find Full Text PDFBiosensors (Basel)
December 2024
School of Health Sciences Research, Research Institute for Health Sciences, Chiang Mai University, Chiang Mai 50200, Thailand.
Benzo[a]pyrene (B[a]P) is a hazardous polycyclic aromatic hydrocarbon that accumulates in several environmental matrices as a result of incomplete combustion. Its presence, carcinogenic properties, and tendency for bioaccumulation provide significant risks to human health and the environment. The objective of this study is to create an immunoassay for the detection of benzo[a]pyrene utilizing immunoglobulin Y antibodies.
View Article and Find Full Text PDFJ Thromb Haemost
December 2024
Division of Hematology, Duke University Medical Center, Durham, NC. Electronic address:
Background: IgG antibodies (Abs) to platelet factor 4 complexed to heparin (PF4/H) commonly occur after heparin exposure but cause life-threatening complications of heparin-induced thrombocytopenia (HIT) in only a few patients. Presently, only platelet activation assays reliably distinguish anti-PF4/H Abs that cause disease (HIT Abs) from those likely to be asymptomatic (AAbs).
Objectives: Recent studies indicate that complement activation is an important serologic property of HIT Abs and is essential for FcγRIIA-mediated cellular activation.
Amino Acids
December 2024
Department of Molecular Biology and Biochemistry, University of California Irvine, Irvine, CA, 92697-3900, USA.
Collapsin response mediator protein 2 (CRMP2) functions in the genesis and activity of neuronal connections in mammalian brain. We previously reported that a protein coincident with CRMP2 on 2D-gels undergoes marked accumulation of abnormal L-isoaspartyl sites in brain extracts of mice missing the repair enzyme, protein L-isoaspartyl methyltransferase (PIMT). To confirm and explore the significance of isoaspartyl damage in CRMP2, we expressed and purified recombinant mouse CRMP2 (rCRMP2).
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