Barnase–barstar high affinity interaction phenomenon as the base for the heterogenous bioluminescence pseudorabies virus' immunoassay

Rudenko, N. V.; Sinegina, Lilia L.; Arzhanov, Maxim A.; Ksenzenko, V. N.; Ивашина, Т. В.; Morenkov, O. S.; Shaloiko, Lyubov; Vinokurov, Leonid M.

doi:10.1016/j.jbbm.2007.01.013

Cited by 5 publications

(4 citation statements)

References 28 publications

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“…Consequently, in the following years a great number of publications described photoprotein-based immunoassays of different analytes of clinical interest – hormones, interleukins, oncomarkers, etc . [12–26], and infections [26–28]. The assays were carried out in different formats (sandwich, competitive), and performed in solid-phase, homogeneous and even flow injection [29] variants.…”

Section: Bioluminescent Immunoassaymentioning

confidence: 99%

Ca2+-Regulated Photoproteins: Effective Immunoassay Reporters

Frank

2010

Sensors

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Ca2+-regulated photoproteins of luminous marine coelenterates are of interest and a challenge for researchers as a unique bioluminescent system and as a promising analytical instrument for both in vivo and in vitro applications. The proteins are comprehensively studied as to biochemical properties, tertiary structures, bioluminescence mechanism, etc. This knowledge, along with available recombinant proteins serves the basis for development of unique bioluminescent detection systems that are “self-contained”, triggerable, fast, highly sensitive, and non-hazardous. In the paper, we focus on the use of photoproteins as reporters in binding assays based on immunological recognition element—bioluminescent immunoassay and hybridization immunoassay, their advantages and prospects.

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Section: Bioluminescent Immunoassaymentioning

confidence: 99%

Ca2+-Regulated Photoproteins: Effective Immunoassay Reporters

Frank

2010

Sensors

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“…. Because of their high binding efficiencies, these complexes are deployed in applications such as nanoparticle biofunctionalisation , construction of supramolecular assemblies and other applications in the area of proteomics and nanoproteomics . They are often used to interface two functional counterparts of a desired bio‐ or nano‐complex, as they are stable in various biomolecular environments .…”

Section: Introductionmentioning

confidence: 99%

“…Several strategies for antibody engineering possible using the Bs:Bn have been reviewed . Although the potential of the Bs:Bn high‑affinity molecular pair in applications such as nanoparticle biofunctionalisation is well recognised , its uptake in the bioassay technologies has been relatively modest . The following two properties of Bs:Bn may address the challenges in the state‐of‐the‐art bioassay technologies, aiding the antigen detection sensitivity progression towards the single‐molecule level.…”

Section: Introductionmentioning

confidence: 99%

A modular design of low‐background bioassays based on a high‐affinity molecular pair barstar:barnase

et al. 2013

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High-affinity molecular pairs provide a convenient and flexible modular base for the design of molecular probes and protein/antigen assays. Specificity and sensitivity performance indicators of a bioassay critically depend on the dissociation constant (K(D)) of the molecular pair, with avidin:biotin being the state-of-the-art molecular pair (K(D) ∼ 1 fM) used almost universally for applications in the fields of nanotechnology and proteomics. In this paper, we present an alternative high-affinity protein pair, barstar:barnase (K(D) ∼ 10 fM), which addresses several shortfalls of the avidin:biotin system, including non-negligible background due to the non-specific binding. A quantitative assessment of the non-specific binding carried out using a model assay revealed inherent irreproducibility of the [strept]avidin:biotin-based assays, attributed to the avidin binding to solid phases, endogenous biotin molecules and serum proteins. On the other hand, the model assays assembled via a barstar:barnase protein linker proved to be immune to such non-specific binding, showing good prospects for high-sensitivity rare biomolecular event nanoproteomic assays.

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“…Desta forma, a presença de luminescência pode ser relacionada à presença de certas moléculas ou organismos, possibilitando sua detecção [73,75,80] . [71][72][73][74][80][81][82][83] .…”

Section: Aplicações Das Terras Raras Como Marcadores Biológicos E Suasunclassified

Marcadores à base de terras raras para detecção de estrogênios sinalizadores em fluídos biológicos

Salum¹

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The development of luminescent, non-toxic and biocompatible nanosensors are being used for marker and imaging of cells in cellular biology and ultrasensitive immunoassays. In this work, biological markers based on rare earths for detection of 17 β-estradiol (E2), in biological fluids (Plasmon), were studied. The precursor complexes based on rare earths (Eu 3+ and Tb 3+) were synthesized and characterized. The complex containing tta Eu 3+ and secondary ligand-TOPO-[Eu(tta) 3 (TOPO) 2 ] registered the highest luminescence efficiency among all other compounds studied and was chosen as luminescent probe for the following development stages of this work. This complex was doped into polymer matrices, PMMA and PHB, at different concentrations of doping by two different methods: cryoscopic nanoprecipitation and emulsion-diffusion technique. The PMMA polymer nanoparticles demonstrated higher luminescence efficiency when doped with 5% concentration of the precursor complex [Eu(tta)3(TOPO)2]. These nanoparticles were functionalized with 1,6 diaminohexane and later they were linked to a dialdehyde that will serve as a bridge in the reaction with the biological entities. These nanoparticles were functionalized with 1,6 diaminohexane and later they were linked to a dialdehyde that can serve as a bridge in the reaction with the biological entities. Due to the energy transfer Ac-Ac/nanosensor, the PHB polymeric nanoparticles showed higher luminescence intensity signal for the detection of 17-βestradiol by spectrofluorimetry. It can be concluded that the nanoparticles produced and tested in this work are strong candidates for the detection of 17β-estradiol in the blood plasma or others biological entities, presenting diagnostic options, an extremely important conduct for therapeutic direction. i LISTA DE FIGURAS FIGURA 1.1-A escala métrica de elementos naturais e dos produzidos pelo homem_________________________________________________________3 FIGURA 3.1-Tabela periódica dos elementos__________________________8 FIGURA 3.2-Representação esquemática (qualitativa) do mecanismo de transferência de energia entre um ligante orgânico e o íon Eu 3+ (CI= Conversão Interna/ CIS= Cruzamento Intersistema/ TE= Transferência de Energia)_____19 FIGURA 3.3-Diagrama de Energia dos íons TR 3+ obtidos a partir de cristais de LaF 3 __________________________________________________________21 FIGURA 3.4-Fórmulas estruturais dos ânions β-dicetonatos ACAC, TTA e DBM__________________________________________________________25 FIGURA 3.5-Estrutura molecular dos ligantes fosfinóxidos: Trifenilfosfinóxido (TPPO) e Tri-n-octilfosfinóxido(TOPO)_______________________________27 FIGURA 3.6-Estrutura molecular do precursor 2-(o-hidroxifenóxi)etil éter e do dibenzo-[18]-coroa-6_____________________________________________27 ii FIGURA 3.7-Estruturas de Éteres Coroa comuns (em destaque: DB18C6 e 18C6)_________________________________________________________29 FIGURA 3.8-Produção mundial de terras raras dos principais países produtores____________________________...

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Barnase–barstar high affinity interaction phenomenon as the base for the heterogenous bioluminescence pseudorabies virus' immunoassay

Cited by 5 publications

References 28 publications

Ca2+-Regulated Photoproteins: Effective Immunoassay Reporters

Ca2+-Regulated Photoproteins: Effective Immunoassay Reporters

A modular design of low‐background bioassays based on a high‐affinity molecular pair barstar:barnase

Marcadores à base de terras raras para detecção de estrogênios sinalizadores em fluídos biológicos

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