Zero-length crosslinking procedure with the use of active esters

Grabarek, Zenon; Gergely, J.

doi:10.1016/0003-2697(90)90267-d

Cited by 756 publications

(604 citation statements)

References 29 publications

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“…1-Ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDC) (Thermo Scientific) and sulfo-NHS (Thermo Scientific)-based heterobifunctional crosslinking of proteins is one of the most widely used procedures for biomolecular conjugation [14][15][16] and biochipbased assay development [17][18][19]. Our results in this study show the interference of sulfo-NHS with the BCA protein assay, which can lead to inaccurate protein estimation.…”

Section: Phospholipids [3] Glucose [4] Mercaptoethanol [4]mentioning

confidence: 97%

“…There are numerous reports where proteins have been quantified by the BCA protein assay in the presence of sulfo-NHS [5][6][7][8][9][10][11][12][13]. However, the interference of sulfo-NHS in the BCA assay has not been reported.1-Ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDC) (Thermo Scientific) and sulfo-NHS (Thermo Scientific)-based heterobifunctional crosslinking of proteins is one of the most widely used procedures for biomolecular conjugation [14][15][16] and biochipbased assay development [17][18][19]. Our results in this study show the interference of sulfo-NHS with the BCA protein assay, which can lead to inaccurate protein estimation.…”

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confidence: 99%

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Sulfo-N-hydroxysuccinimide interferes with bicinchoninic acid protein assay

Vashist

Zhang

Zheng

et al. 2011

Analytical Biochemistry

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This study revealed a major interference from sulfo-N-hydroxysuccinimide (sulfo-NHS) in the bicinchoninic acid (BCA) protein assay. Sulfo-NHS, a common reagent used in bioconjugation and analytical biochemistry, exhibited absorbance signals and absorbance peaks at 562 nm, comparable to bovine serum albumin (BSA). However, the combined absorbance of sulfo-NHS and BSA was not strictly additive. The sulfo-NHS interference was suggested to be caused by the reduction of Cu 2+ in the BCA Kit's reagent B (4% cupric sulfate) in a manner similar to that of the protein.Crown Copyright Ó 2011 Published by Elsevier Inc. All rights reserved.The bicinchoninic acid (BCA) 1 protein assay [1] is widely used for determination of protein concentrations ranging from 25 to 2000 lg/mL. As a water-soluble sodium salt, BCA exhibits high sensitivity and specificity for Cu 1+ . Similar to the Biuret reaction, the presence of protein in an alkaline medium reduces Cu 2+ to Cu 1+ . Consequently, a purple-colored reaction product is formed by the chelation of two molecules of BCA with each Cu 1+ , which exhibits a strong absorbance at 562 nm. The total protein concentration is reflected by the sample color change from green to purple in proportion to a given protein concentration. The color formation caused by the reaction of protein with BCA is dependent on its number of peptide bonds, its macromolecular structure, and the availability of four amino acids (cysteine, cystine, tryptophan, and tyrosine) in the protein [2]. The amino acid sequence, pI, and the presence of certain side chains or prosthetic groups can also affect the color formation significantly. The extent of the color formation is determined by more than the sum of individual color-forming functional groups [2].The Thermo Scientific BCA protein assay kit's instruction (Cat. No. 23227) indicates several potential interfering substances such as ascorbic acid, catecholamine, creatinine, cysteine, EGTA, impure glycerol, hydrogen peroxide, hydrazides, iron, lipids, melibiose, phenol red, impure sucrose, tryptophan, tyrosine, and uric acid. Phospholipids [3], glucose [4], mercaptoethanol [4], and other substances such as acetamidophenol, 3,4-dihydroxyphenylalanine, dithiothreitol, glutathione, and penicillamine [2] also interfere with the BCA assay. There is also a temperature-dependent reaction of peptide bonds with Cu 2+ that can increase the concentration of Cu 1+ . There are numerous reports where proteins have been quantified by the BCA protein assay in the presence of sulfo-NHS [5][6][7][8][9][10][11][12][13]. However, the interference of sulfo-NHS in the BCA assay has not been reported.1-Ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDC) (Thermo Scientific) and sulfo-NHS (Thermo Scientific)-based heterobifunctional crosslinking of proteins is one of the most widely used procedures for biomolecular conjugation [14][15][16] and biochipbased assay development [17][18][19]. Our results in this study show the interference of sulfo-NHS with the BCA protein assay, which can ...

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Section: Phospholipids [3] Glucose [4] Mercaptoethanol [4]mentioning

confidence: 97%

mentioning

confidence: 99%

Sulfo-N-hydroxysuccinimide interferes with bicinchoninic acid protein assay

Vashist

Zhang

Zheng

et al. 2011

Analytical Biochemistry

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“…37 At low pH, EDCactivated carbodiimide attachment rates will decrease and EDC decomposition will increase. 34 Other electrochemical decomposition reactions 38−40 may also occur at the electrodes. Typically reduced voltage, 27,41,42 pulsed dc fields, 37,42−44 and asymmetric ac fields 44,45 have previously been found to inhibit electrolysis.…”

Section: ■ Results and Discussionmentioning

confidence: 99%

Directing Cluster Formation of Au Nanoparticles from Colloidal Solution

et al. 2013

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Discrete clusters of closely spaced Au nanoparticles can be utilized in devices from photovoltaics to molecular sensors because of the formation of strong local electromagnetic field enhancements when illuminated near their plasmon resonance. In this study, scalable, chemical self-organization methods are shown to produce Au nanoparticle clusters with uniform nanometer interparticle spacing. The performance of two different methods, namely electrophoresis and diffusion, for driving the attachment of Au nanoparticles using a chemical cross-linker on chemically patterned domains of polystyreneblock-poly(methyl methacrylate) (PS-b-PMMA) thin films are evaluated. Significantly, electrophoresis is found to produce similar surface coverage as diffusion in 1/6th of the processing time with an ∼2-fold increase in the number of Au nanoparticles forming clusters. Furthermore, average interparticle spacing within Au nanoparticle clusters was found to decrease from 2−7 nm for diffusion deposition to approximately 1−2 nm for electrophoresis deposition, and the latter method exhibited better uniformity with most clusters appearing to have about 1 nm spacing between nanoparticles. The advantage of such fabrication capability is supported by calculations of local electric field enhancements using electromagnetic full-wave simulations from which we can estimate surface-enhanced Raman scattering (SERS) enhancements. In particular, full-wave results show that the maximum SERS enhancement, as estimated here as the fourth power of the local electric field, increases by a factor of 100 when the gap goes from 2 to 1 nm, reaching values as large as 10 10 , strengthening the usage of electrophoresis versus diffusion for the development of molecular sensors.

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“…Besides the coupling procedures using EDC alone, EDC together with N-hydroxysulfosuccinimide sodium salt (sulfo-NHS) as cross-linker can be added. Sulfo-NHS is known to enhance the coupling efficiency by preparing amine-reactive esters of carboxylate groups (stabilizes the unstable ester O-acylisourea [12] (Table 6).…”

Section: Design Of Mnps As Multifunctional Tools For Magnetic Hyperthmentioning

confidence: 99%

Means to increase the therapeutic efficiency of magnetic heating of tumors

Kettering

Grau

Pömpner

et al. 2015

Biomedical Engineering / Biomedizinische Technik

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Abstract:The treatment of tumors via hyperthermia has gained increased attention in the last years. Among the different modalities available so far, magnetic hyperthermia has the particular advantage of offering the possibility of depositing the heating source directly into the tumor. In this study, we summarized the present knowledge we gained on how to improve the therapeutic efficiency of magnetic hyperthermia using magnetic nanoparticles (MNPs), with particular consideration of the intratumoral infiltration of the magnetic material. We found that (1) MNPs will be mainly immobilized at the tumor area and that this aspect has to be considered when estimating the heating potential of MNPs, (2) the intratumoral distribution patterns via slow infiltration might well be modulated by specific MNP coating and magnetic targeting, (3) imaging of the nanoparticle depositions within the tumor might allow to correct the distribution pattern via multiple applications, (4) multiple therapeutic sessions are feasible because MNPs are not delivered from the tumor site during the heating process, (5) the utilization of MNPs that internalize into cells will favor the production of intracellular heating spots rather than extracellular ones, (6) utilization of MNPs functionalized with chemotherapeutic agents will allow us to exploit the additive effects of both therapeutic modalities, and (7) distinct cytopathological and histopathological alterations in target tissues are induced as a result of magnetic hyperthermia. However, the accumulation at the tumor via intravenous application remains a matter of challenge.

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Zero-length crosslinking procedure with the use of active esters

Cited by 756 publications

References 29 publications

Sulfo-N-hydroxysuccinimide interferes with bicinchoninic acid protein assay

Sulfo-N-hydroxysuccinimide interferes with bicinchoninic acid protein assay

Directing Cluster Formation of Au Nanoparticles from Colloidal Solution

Means to increase the therapeutic efficiency of magnetic heating of tumors

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