Development of ELISA formats for polymyxin B monitoring in serum of critically ill patients

Burkin, Maksim A.; Galvidis, Inna A.; Surovoy, Yury A.; Plyushchenko, I. V.; Родин, И. А.; Царенко, С. В.

doi:10.1016/j.jpba.2021.114275

Cited by 14 publications

(7 citation statements)

References 24 publications

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“…Since the aecAZI hapten conjugated to the carrier via a zerolength arm yielded the desired assay performance, we prepared the same design enzymelabeled hapten for the direct competitive assay format. Consistent with our previous observation [21], binding an enzyme-labeled hapten with a lower molar ratio between hapten and HRP (3:1) resulted in better analyte competition and assay sensitivity compared to a 10:1 ratio. Thus, a direct competitive assay format was optimized and established based on coated anti-BSA-cmoCLA(ae) antibody and HRP-aecAZI × 3 as a tracer.…”

Section: Direct Competitive Assay Formatssupporting

confidence: 91%

“…Hapten conjugation to HRP was conducted using the periodate method as in a previous work [21]. Briefly, HRP solution (3.2 mg, 80 nmol) in 0.4 mL H 2 O was combined with an equal volume of sodium periodate (1.7 mg, 8 µmol) and stirred for 15 min with a magnet stirrer.…”

Section: Hrp-labeled Antigen Preparationmentioning

confidence: 99%

“…The antibody coated on polystyrene plates could capture the free analyte from the tested sample and enzymelabeled hapten in a competitive manner. The sequence of manipulations corresponded to those described for a similar format earlier [21]. The role of the volume ratios between the standard (25-275 µL) and HRP-aecAZI (25-175 µL) on the sensitivity of the assay was studied in this work.…”

Section: Direct Competitive Assay Formatsmentioning

confidence: 99%

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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

Preprint

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Macrolide antibiotics, effective antimicrobial agents, are intensively used in human and veterinary medicine as well as in agriculture and therefore are found all over the world as environmental pollutants, harming sensitive eco-community organisms and provoking the selection of resistant forms. A novel azithromycin derivative was synthesized and as a rationally designed hapten conjugate ensured group immunorecognition of 6 major macrolide representatives (105-41%), erythromycin, erythromycin ethylsuccinate, clarithromycin, roxithromycin, azithromycin and dirithromycin in competitive immunoassay based on anti-clarithromycin antibodies. The heterologous hapten-based ELISA format simultaneously contributed to a 5-fold increase in sensitivity (ERY IC50 = 0.04 ng/mL). However, for the detection of trace macrolides in environmental samples, an underexploited in immunoassay field strategy was proposed in the present study to significantly improve the detectability of analytes. Unlike most approaches, it does not require special enhancers/amplifiers or additional concentration/extraction procedures, but only involves a larger volume of test samples. Gradual volume increase of samples (from 0.025 to 10 mL) analyzed in 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 macrolide contamination of natural water was confirmed by recovery of macrolides from spiked blank samples (71.7-141.3%). A series of natural water samples from Lake Onega and its influents near Petrozavodsk were analyzed during a 2022-2023 using both the developed immunoassay and HPLC-MS/MS and revealed no macrolide antibiotic contamination.

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Section: Direct Competitive Assay Formatssupporting

confidence: 91%

Section: Hrp-labeled Antigen Preparationmentioning

confidence: 99%

Section: Direct Competitive Assay Formatsmentioning

confidence: 99%

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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

Preprint

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“…Colistin MS is a prodrug form with blocked amine groups, so it was poorly recognized by the antibody. However, as a result of hydrolysis in vivo, CMS was converted into COL and could be detected as active drug [ 32 ].…”

Section: Resultsmentioning

confidence: 99%

Influence of Endogenous Factors of Food Matrices on Avidin—Biotin Immunoassays for the Detection of Bacitracin and Colistin in Food

Burkin

Galvidis

Eremin

2022

Foods

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(Strept)avidin–biotin technology is frequently used in immunoassay systems to improve their analytical properties. It is known from clinical practice that many (strept)avidin–biotin-based tests provide false results when analyzing patient samples with a high content of endogenous biotin. No specific investigation has been carried out regarding possible interferences from avidin (AVI) and biotin (B7) contained in food matrices in (strept)avidin–biotin-based immunoanalytical systems for food safety. Two kinds of competitive ELISAs for bacitracin (BT) and colistin (COL) determination in food matrices were developed based on conventional hapten–protein coating conjugates and biotinylated BT and COL bound to immobilized streptavidin (SAV). Coating SAV–B7–BT and SAV–B7–COL complexes-based ELISAs provided 2- and 15-times better sensitivity in BT and COL determination, corresponding to 0.6 and 0.3 ng/mL, respectively. Simultaneously with the determination of the main analytes, these kinds of tests were used as competitive assays for the assessment of AVI or B7 content up to 10 and 1 ng/mL, respectively, in food matrices (egg, infant milk formulas enriched with B7, chicken and beef liver). Matrix-free experiments with AVI/B7-enriched solutions showed distortion of the standard curves, indicating that these ingredients interfere with the adequate quantification of analytes. Summarizing the experience of the present study, it is recommended to avoid immunoassays based on avidin–biotin interactions when analyzing biosamples containing these endogenous factors or enriched with B7.

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“…Polymyxin B concentration in tested samples was measured using a direct competitive ELISA described in detail previously [23]. The assay detection limit (IC 10 ) was 1.8 ng/mL, the dynamic range (IC 20 -IC 80 ) constituted 5-200 ng/mL.…”

Section: Elisa Proceduresmentioning

confidence: 99%

Comparative polymyxin B pharmacokinetics in critically ill patients with renal insufficiency and in continuous veno-venous hemodialysis

Surovoy

Burkin

Galvidis

et al. 2022

Eur J Clin Pharmacol

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The aim of this study was to assess polymyxin B pharmacokinetics (PK) in patients with varying degrees of renal dysfunction and in patients, who require continuous veno-venous hemodialysis (CVVHD). The study enrolled 37 patients with sepsis, among whom 13 patients with glomerular ltration rate below 80 ml/min and 11 patients on CVVHD. For every patient 6-8 blood samples were collected during 12-hour dosage interval. Polymyxin B serum concentration was determined using enzyme-linked immunosorbent assay. In sepsis patients with preserved renal function mean area under the curve over 24 hours (AUC 0 − 24h ) value reached 67.8 ± 9.8 mg*h/L, while in patients with glomerular ltration rate (GFR) below 80 ml/min mean AUC 0 − 24h was 87 ± 5.8 mg*h/L. PMB PK in patients with renal insu ciency was characterized by signi cantly lower clearance (CL) compared to normal renal function group (2.1 ± 0.1 L/h vs 3.9 ± 0.4 L/h respectively). In patients on CVVHD mean AUC 0 − 24h was 110.4 ± 10.3 mg*h/L, while CL reached 2 ± 0.23 L/h. Median recovery rate from dialysate constituted 22%. Simulation of different dosage regimens indicate xed maintenance dose of 100 mg q12h is optimal for patients on CVVHD and no dosage increase is required. This study demonstrates decreased clearance of PMB in patients with renal insu ciency, which puts them at risk of toxicity. Therefore, patients with extremes of renal function might bene t from therapeutic drug monitoring. For patients with anuria, who require CVVHD, we suggest a xed dose of 100 mg q12h. Key PointsAcute kidney injury has signi cant impact on polymyxin B clearance, and therapeutic drug monitoring might be bene cial in certain groups of critically ill patients.Continuous veno-venous hemodialysis does not lead to signi cant removal of polymyxin B, therefore a xed dosage regimen of 100 mg twice daily might be appropriate in these patients.

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Development of ELISA formats for polymyxin B monitoring in serum of critically ill patients

Cited by 14 publications

References 24 publications

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

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

Influence of Endogenous Factors of Food Matrices on Avidin—Biotin Immunoassays for the Detection of Bacitracin and Colistin in Food

Comparative polymyxin B pharmacokinetics in critically ill patients with renal insufficiency and in continuous veno-venous hemodialysis

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