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 20 publications

(7 citation statements)

References 24 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…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

Self Cite

View full text Add to dashboard Cite

(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.

show abstract

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

Self Cite

View full text Add to dashboard Cite

show abstract

“…This type of assay does not require the preprocessing of paenibacillin-containing samples (i.e., crude or impure extracts can be used directly) and is less sensitive to matrix inhibitory effects. Competitive ELISAs have been successfully developed for several antimicrobial peptides, such as polymyxin B and nisin [12,[27][28][29][30]. In the current study, a sigmoidal three-parameter nonlinear regression model was used to construct a competitive ELISA standard curve, which accurately correlated paenibacillin concentration with the OD 450 signals (R 2 = 0.95).…”

Section: Discussionmentioning

confidence: 98%

Production of Polyclonal Antibodies and Development of Competitive ELISA for Quantification of the Lantibiotic Paenibacillin

Abdelhamid,

Wick,

Yousef

2024

Fermentation

View full text Add to dashboard Cite

The discovery and biotechnological application of new antimicrobial peptides are impeded by a lack of sensitive methods for peptide quantification. Paenibacillin is an emerging antimicrobial lantibiotic that was discovered in Paenibacillus polymyxa OSY-DF ATCC PTA-7852, isolated from the fermented vegetable Kimchee. This lantibiotic has potency against many foodborne pathogenic and spoilage bacteria. To advance the research and application of paenibacillin, a rapid, specific, and sensitive detection and quantification immunoassay was developed. After anti-paenibacillin polyclonal antibodies (pAbs) were generated and purified, a competitive enzyme-linked immunosorbent assay (cELISA) was developed and optimized for paenibacillin quantification. The dynamic range of the cELISA was determined by using a three-parameter nonlinear regression model, achieving a correlation (R2) value of 0.95. The cELISA displayed high sensitivity, with the ability to detect paenibacillin at levels as low as 15.6 ng/mL, which is significantly lower than the limit of detection of the conventional antimicrobial assay (20 µg/mL paenibacillin). The cELISA successfully differentiated paenibacillin concentrations in cell-free crude supernatants of P. polymyxa wild type and its mutant strain when grown at 30 °C and 37 °C; higher paenibacillin levels were found in the mutant (0.248–0.276 µg/mL) than in the wild type (0.122–0.212 µg/mL) culture. These findings were validated by the transcriptional analysis of 11 paenibacillin biosynthetic genes, which were significantly upregulated (≥2-fold increase) in the mutant compared with the wild strain. Additionally, the cELISA exhibited high sensitivity by recovery of paenibacillin titers spiked at 2.5 and 10 µg/mL in de Man, Rogosa, and Sharpe (MRS) broth and diluted skim milk. These results suggest that the anti-paenibacillin pAbs and the developed cELISA could be valuable in quantifying paenibacillin in complex matrices and in aiding the discovery of paenibacillin-producing natural microbiota.

show abstract

“…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%

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

Self Cite

View full text Add to dashboard Cite

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.

show abstract

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

Cited by 20 publications

References 24 publications

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

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

Production of Polyclonal Antibodies and Development of Competitive ELISA for Quantification of the Lantibiotic Paenibacillin

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

Contact Info

Product

Resources

About