Dataset Information

Synthesis and Structural Optimization of Iridium(III) Solvent Complex for Electrochemiluminescence Labeling of Histidine-Rich Protein and Immunoassay Applications for CRP Detection.

ABSTRACT: The reaction between an iridium(III) solvent complex and the histidine site of biomolecules as one kind of novel bioconjugation approaches has received much attention during the past few years. To extend this novel bioconjugation approach into electrochemiluminescence (ECL) immunoassay and optimize the performances, three iridium(III) solvent complexes with different C?N bidentate main ligands have been designed and synthesized in this work. Bovine serum albumin (BSA) as the standard histidine-rich protein is initially employed to evaluate the labeling performances by comparing the ECL intensity of the same amount of BSA labeled by different iridium(III) solvent complexes. Importantly, a magnetic beads-based sandwich immunoassay platform using Ir-dmpq (iridium(III) acetonitrile complex with 2-(3,5-dimethylphenyl)quinoline as the main ligand) as a structurally optimized labeling agent has been successfully constructed to detect C-reactive protein (CRP, an important biomarker of systemic inflammation in clinic), and the limit of detection based on this novel labeling agent could reach below 1 ng/mL, which may further pave the way for applications of the iridium(III) solvent complex in histidine-rich protein ECL labeling beyond fluorescence labeling.

SUBMITTER: Zhou Y

PROVIDER: S-EPMC7045565 | biostudies-literature | 2020 Feb

REPOSITORIES: biostudies-literature

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Synthesis and Structural Optimization of Iridium(III) Solvent Complex for Electrochemiluminescence Labeling of Histidine-Rich Protein and Immunoassay Applications for CRP Detection.

Zhou Yuyang Y Ding Yangming Y Huang Yinhao Y Cai Leilei L Xu Jiawei J Ma Xiao X

ACS omega 20200211 7

The reaction between an iridium(III) solvent complex and the histidine site of biomolecules as one kind of novel bioconjugation approaches has received much attention during the past few years. To extend this novel bioconjugation approach into electrochemiluminescence (ECL) immunoassay and optimize the performances, three iridium(III) solvent complexes with different C<sup>∧</sup>N bidentate main ligands have been designed and synthesized in this work. Bovine serum albumin (BSA) as the standard ...[more]

PMID: 32118179

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Project description:BackgroundCyclosporine A (CsA) is used as a posttransplantation immunosuppressant drug, and careful monitoring of CsA concentration in whole blood is essential. A new automated electrochemiluminescence immunoassay (ECLIA) for CsA measurement has been assessed in a multicenter evaluation.MethodsResidual EDTA whole blood samples from patients undergoing CsA therapy after organ transplant were used in assay evaluation at 5 clinical laboratories in Europe. Experiments included imprecision according to CLSI EP5-A2 (within-run and intermediate), lower limit of quantification, linearity according to CLSI EP6-A, and recovery of commercial external quality control samples. In addition, comparisons to liquid chromatography-tandem mass spectrometry methods in routine use at each investigational site and to commercial chemiluminescent microparticle immunoassay and antibody-conjugated magnetic immunoassay methods were performed.ResultsImprecision testing gave coefficients of variation of less than 9% in the 30-2000 mcg/L range for both within-run and intermediate imprecision. Lower limit of quantification of 6.8 mcg/L at one investigational site and 1.8 mcg/L at a second site at 20% coefficient of variation were observed. Linearity was measured over the concentration range 0-2000 mcg/L, yielding a deviation of less than ±12%. External quality control sample recovery by ECLIA was 93%-114% of LC-MS/MS sample recovery. Deming regression analysis of ECLIA method comparison to combined LC-MS/MS results yielded a slope of 1.04 [95% confidence interval (CI), 1.03-1.06] and intercept of 2.8 mcg/L (95% CI, 1.5-4.1 mcg/L). Comparison to chemiluminescent microparticle immunoassay yielded a slope of 0.87 (95% CI, 0.85-0.89) and intercept of 1.4 mcg/L (95% CI, -0.89 to 3.7 mcg/L); comparison to antibody-conjugated magnetic immunoassay yielded a slope of 0.96 (95% CI, 0.93-0.98) and intercept of -4.2 mcg/L (95% CI, -7.1 to -1.2 mcg/L).ConclusionsThe data from this multicenter evaluation indicate that the new ECLIA-based cyclosporine assay is fit for its purpose, the therapeutic monitoring of CsA.

Project description:BACKGROUND:Malaria rapid diagnostic tests (RDTs) are precious tools to diagnose malaria. Most RDTs used currently are based on the detection of Plasmodium falciparum histidine-rich protein 2 (PfHRP2) in a patient's blood. However, concern has been raised in recent years that deletion of pfhrp2 in the parasite could affect the accuracy of PfHRP2-based RDTs. In addition, genetic variation in pfhrp2 might influence the accuracy and sensitivity of RDTs. In this study, the genetic variation in pfhrp2 and pfhrp3 in Myanmar P. falciparum isolates was analysed. METHODS:Blood samples were collected from malaria patients who were infected with P. falciparum in Mandalay, Naung Cho, Tha Beik Kyin, and Pyin Oo Lwin, Upper Myanmar between 2013 and 2015. The pfhrp2 and pfhrp3 were amplified by nested polymerase chain reaction (PCR), cloned and sequenced. Genetic variation in Myanmar pfhrp2 and pfhrp3 was analysed using the DNASTAR program. Comparative analysis of Myanmar and global pfhrp2 and pfhrp3 isolates was also performed. RESULTS:One-hundred and two pfhrp2 and 89 pfhrp3 were amplified from 105 blood samples, of which 84 pfhrp2 and 56 pfhrp3 sequences were obtained successfully. Myanmar pfhrp2 and pfhrp3 showed high levels of genetic variation with different arrangements of distinct repeat types, which further classified Myanmar pfhrp2 and pfhrp3 into 76 and 47 haplotypes, respectively. Novel amino acid changes were also found in Myanmar pfhrp2 and pfhrp3, but their frequencies were very low. Similar structural organization was shared by Myanmar and global pfhrp2 and pfhrp3, and differences in frequencies of repeat types and lengths were also observed between and among global isolates. CONCLUSION:Length polymorphisms and amino acid substitutions generated extensive genetic variation in Myanmar pfhrp2 and pfhrp3. Comparative analysis revealed that global pfhrp2 and pfhrp3 share similar structural features, as well as extensive length polymorphisms and distinct organizations of repeat types. These results provide a better understanding of the genetic structure of pfhrp2 and pfhrp3 in global P. falciparum populations and suggest useful information to develop RDTs with improved quality.

Dataset Information

Synthesis and Structural Optimization of Iridium(III) Solvent Complex for Electrochemiluminescence Labeling of Histidine-Rich Protein and Immunoassay Applications for CRP Detection.

Publications

Synthesis and Structural Optimization of Iridium(III) Solvent Complex for Electrochemiluminescence Labeling of Histidine-Rich Protein and Immunoassay Applications for CRP Detection.

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