Advances in the study of luminescence probes for proteins

Sun, Cheng; Yang, Jinghe; Li, Lei; Wu, Xia; Liu, Yang; Liu, Shufang

doi:10.1016/j.jchromb.2003.12.039

Cited by 103 publications

(46 citation statements)

References 175 publications

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“…Dye-binding methods are widely used for the determination of proteins, 13) and spectrophotometric methods using bromocresol green are most commonly used in clinical assays of human serum albumin (HSA). [14][15][16][17] However, the specificity for HSA in the bromocresol green method is not very high.…”

mentioning

confidence: 99%

Fluorimetric Determination of Trace Amounts of Albumin in Bronchoalveolar Lavage Fluid with Eriochrome Cyanine R

Sato

Saito

Chikuma

et al. 2007

Biological & Pharmaceutical Bulletin

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Bronchoalveolar lavage has been widely used to recover cells, proteins, and small molecules from the alveolar space of the lower respiratory tract since the first report of Reynolds and Newball over three decades ago.1) The analysis of these components is of wide diagnostic and investigative value. [1][2][3][4][5][6][7][8][9][10][11][12] In bronchoalveolar lavage, sterile saline is administered into the bronchoalveolar space and aspirated by syringe. 1)Because recovery of the components in bronchoalveolar lavage fluid (BALF) varies from sample to sample, their concentrations are usually normalized relative to those of albumin. [1][2][3][4][5][6]8) Furthermore, the absolute concentration of albumin itself in BALF may allow characterization of interstitial lung disorders.9,10) Therefore a sensitive and simple method for determining trace amounts of albumin in BALF is desirable.Dye-binding methods are widely used for the determination of proteins, 13) and spectrophotometric methods using bromocresol green are most commonly used in clinical assays of human serum albumin (HSA).14-17) However, the specificity for HSA in the bromocresol green method is not very high. The bromocresol purple method is also used in clinical measurement of HSA, and is more selective for HSA than the bromocresol green method. 18,19) However, the bromocresol purple method is not applicable to BALF samples of low albumin concentration.Saito et al. 20,21) reported a highly sensitive fluorimetric method using chromazurol S (CAS) for the determination of trace albumin, and pointed out that several reagents analogous to CAS including eriochrome cyanine R (ECR) are promising.22) Subsequently, Ci and Chen 23) reported a fluorimetric method for the determination of HSA using ECR. They used 308 nm as the excitation wavelength. However, Saito's group reported that the HSA-ECR complex showed visible excitation and emission spectra and low reagent blank at pH <4.0. Ultraviolet radiation may give rise to fluorescence of other components. Therefore in the present study we investigated the fluorimetric determination method of human albumin using ECR and visible range as excitation wavelength. This technique was applied to determine trace amounts of human albumin in BALF samples. MATERIALS AND METHODS ReagentsThe chemicals used in the present study were obtained from the sources indicated as follows: Eriochrome cyanine R (ECR), Merck AG (Darmstadt, Germany); human serum albumin (HSA), fraction V grade, and goat anti-HSA antiserum, Sigma Chemical Co. (St. Louis, U.S.A.); 3,3Ј,5,5Ј-tetramethylbenzidine, Nacalai Tesque Inc. (Kyoto, Japan). All solutions were prepared using water purified with a Milli-Q water system (Millipore Corp., Bedford, U.S.A.). Polystyrene balls were obtained from Sekisui Chemical Co., Ltd. (Tokyo, Japan). Bronchoalveolar Lavage Informed consent was obtained from all patients. Fifty milliliters of warm saline was infused through the work channel of a bronchoscope then withdrawn by gentle aspiration. The lavage was repeated 6 times, and a...

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confidence: 99%

Fluorimetric Determination of Trace Amounts of Albumin in Bronchoalveolar Lavage Fluid with Eriochrome Cyanine R

Sato

Saito

Chikuma

et al. 2007

Biological & Pharmaceutical Bulletin

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“…[29] This is in contrast to the number of amino groups: HSA contains 60 lysines, 35 of which are freely accessible. [30] Labeling of amino groups not only occurs randomly, [31] but may also lead to a range of DPRs, whilst thiol labeling of HSA results in highly sitespecific and strictly single labeling as demonstrated by a DPR of 1. Thiol labeling also eliminates the risk of selfquenching of fluorophores due to multiple labeling.…”

Section: Discussionmentioning

confidence: 99%

Click Chemistry Based Method for the Preparation of Maleinimide‐Type Thiol‐Reactive Labels

Link

Kleim

et al. 2010

Eur J Org Chem

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Maleinimides are widely used to label proteins and to derivatize thiols. The so‐called click reaction was shown to allow the efficient introduction of the maleinimido group into azido‐modified fluorophores (benzoxazines) by reacting them with an alkyne‐modified maleinimide to yield new thiol‐reactive fluorescent labels 5–7. The reaction proceeds under mild experimental conditions and provides a new strategy with which to introduce the maleimide group, as exemplified here for fluorophores. Conceivably, it may be extended to radioactive, electro‐active, isotopic, or spin labels. Previously reported methods for the formation of such maleinimides starting from open‐ring precursors require rather harsh conditions. The new benzoxazines 5–7 are characterized by a fairly long and flexible linker between the chromophore and the maleinimide functional group, and their fluorescence can be photoexcited with diode lasers (which are preferred light sources in fluorometry). They were conjugated to: (a) the aminothiol cysteamine, (b) the peptide glutathione, and (c) to human serum albumin. The fluorescence of the phenoxazinone 5 is strongly solvatochromic, which suggests its use as a polarity‐sensitive probe.

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“…This detection problem could be solved by derivatization procedures which allow transforming amino acids onto detectable forms. Many publications have reported the use of derivatization methods for improving a protein's detectability [11].…”

Section: Introductionmentioning

confidence: 99%

Validation of a HPLC Method for Determination of Glutamine in Food Additives Using Post-Column Derivatization

Chorilli¹,

Salgado²,

Santos³

et al. 2012

AJAC

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An ion-exchange chromatography (IEC) followed by post-column derivatization method was validated for the determination of glutamine in food additive products. The chromatographic method was carried out on a sodium cation-exchange column and using detection at 570 nm. The mobile phase consisted of sodium eluant, pH 3.15, with 5% sulfolane, sodium eluant, pH 7.40 and sodium column regenerant. The method was linear in the range of 4 -50 µg/mL (r 2 = 0.9999). The accuracy was 99.78%. Moreover, method validation demonstrated acceptable results for precision and robustness. The method was found to be able to use for routine analysis of the glutamine.

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Advances in the study of luminescence probes for proteins

Cited by 103 publications

References 175 publications

Fluorimetric Determination of Trace Amounts of Albumin in Bronchoalveolar Lavage Fluid with Eriochrome Cyanine R

Fluorimetric Determination of Trace Amounts of Albumin in Bronchoalveolar Lavage Fluid with Eriochrome Cyanine R

Click Chemistry Based Method for the Preparation of Maleinimide‐Type Thiol‐Reactive Labels

Validation of a HPLC Method for Determination of Glutamine in Food Additives Using Post-Column Derivatization

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