Three steps improving the sensitivity of sulfonamide immunodetection in milk

Burkin, Maksim A.; Lapa, G. B.; Galvidis, Inna A.; Burkin, Konstantin M.; Zubkov, A. V.; Eremin, Sergei A.

doi:10.1039/c8ay01904e

Cited by 6 publications

(2 citation statements)

References 26 publications

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“…Indirect antigen-coated enzyme-linked immunoassay was executed according to the procedure described before [32]. Optimal reagent concentrations were determined using check board titration.…”

Section: Methodsmentioning

confidence: 99%

Specific and Generic Immunorecognition of Glycopeptide Antibiotics Promoted by Unique and Multiple Orientations of Hapten

2019

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Conjugation chemistry does not always provide adequate spatial orientation of hapten in immunogens for the best presentation of generic or individual epitopes. In the present study, the influence of unique and multiple orientations of immunizing hapten on the immune response repertoire was compared to select generic recognition system. The glycopeptides, teicoplanin (TPL) and ristomycin (RSM), were conjugated to BSA to produce immunogens with unique and multiple orientations of haptens. Polyclonal antibodies generated against TPL conjugated through a single site were of uniform specificity and demonstrated selective TPL recognition, regardless of the coating conjugates design. The sensitivity (IC50) of 4 enzyme-linked immunosorbent assays (ELISAs) for TPL varied little within the 3.5–7.4 ng/mL, with a dynamic range of 0.2–100 ng/mL. RSM was coupled to BSA through several glycoside sites that evoked a wider repertoire of response. This first described anti-RSM antibody was selective for RSM in homologous hapten-coated ELISAs with IC50 values in the range 4.2–35 ng/mL. Among the heterologous antigens, periodate-oxidized TPL conjugated to gelatine was selected as the best binder of generic anti-RSM fraction. The developed ELISA showed group recognition of glycopeptides RSM, TPL, eremomycin, and vancomycin with cross-reactivity of 37–100% and a 10–10,000 ng/mL dynamic range. Thus, multiple presentations of immunizing hapten help expand the repertoire of immune responses and opportunities for the selection of the required fine-specificity agent.

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Section: Methodsmentioning

confidence: 99%

Specific and Generic Immunorecognition of Glycopeptide Antibiotics Promoted by Unique and Multiple Orientations of Hapten

2019

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“…Thus, the approach using a heterologous hapten aecAZI was one of the steps taken to improve the assay sensitivity [32]. Subsequently, by utilizing only the internal resource of the prepared reagents without additional labels, enhancers, or amplifiers, the sensitivity of the assay can be significantly increased by a larger sample volume, surpassing the sensitivity of many reported assays (Table 2).…”

Section: Effect Of Sample Volume Increase On Sensitivity Assessed In ...mentioning

confidence: 99%

Immunotechniques for the Group Determination of Macrolide Antibiotics Traces in the Environment Using a Volume-Mediated Sensitivity Enhancement Strategy

Burkin,

Tevyashova,

Bychkova

et al. 2023

Biosensors

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Macrolide antibiotics, which are effective antimicrobial agents, are intensively used in human and veterinary medicine, as well as in agriculture. Consequently, they are found all over the world as environmental pollutants, causing harm to sensitive ecological communities and provoking a selection of resistant forms. A novel azithromycin derivative, which was used as hapten conjugate, ensured the group immunorecognition of six major macrolide representatives (105–41%), namely erythromycin, erythromycin ethylsuccinate, clarithromycin, roxithromycin, azithromycin, and dirithromycin in a competitive immunoassay based on anti-clarithromycin antibodies. The heterologous hapten-based ELISA format resulted in a 5-fold increase in sensitivity, with an IC50 value of 0.04 ng/mL for erythromycin. In this study, we proposed an underexploited strategy in an immunoassay field to significantly improve the detectability of analytes in environmental samples. Unlike most approaches, it does not require special enhancers/amplifiers or additional concentration/extraction procedures; instead, it involves analyzing a larger volume of test samples. A gradual volume increase in the samples (from 0.025 to 10 mL) analyzed using a direct competitive ELISA, immunobeads, and immunofiltration assay formats based on the same reagents resulted in a significant improvement (more than 50-fold) in assay sensitivity and detection limit up to 5 and 1 pg/mL, respectively. The suitability of the test for detecting the macrolide contamination of natural water was confirmed by the recovery of macrolides from spiked blank samples (71.7–141.3%). During 2022–2023, a series of natural water samples from Lake Onega and its influents near Petrozavodsk were analyzed, using both the developed immunoassay and HPLC-MS/MS. The results revealed no contamination of macrolide antibiotic.

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Production of generic monoclonal antibody and development of chemiluminescence immunoassay for determination of 32 sulfonamides in chicken muscle

Cui

Liu

et al. 2020

Food Chemistry

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Three steps improving the sensitivity of sulfonamide immunodetection in milk

Abstract: Three approaches improving the sensitivity of previously established immunoassays for broad detection of sulfonamides (SAs) in milk are described.

Cited by 6 publications

References 26 publications

Specific and Generic Immunorecognition of Glycopeptide Antibiotics Promoted by Unique and Multiple Orientations of Hapten

Specific and Generic Immunorecognition of Glycopeptide Antibiotics Promoted by Unique and Multiple Orientations of Hapten

Immunotechniques for the Group Determination of Macrolide Antibiotics Traces in the Environment Using a Volume-Mediated Sensitivity Enhancement Strategy

Production of generic monoclonal antibody and development of chemiluminescence immunoassay for determination of 32 sulfonamides in chicken muscle

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