Zero-Length Crosslinkers

Hermanson, Greg T.

doi:10.1016/b978-0-12-382239-0.00004-2

Cited by 112 publications

(113 citation statements)

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“…We then added 1 mL of EDC (at 0.4 mg⋅mL −1 ) and, after 5 minutes, 1 mL of Sulfo-NHS at 1.1 mg⋅mL −1 to the QDs sample. 7,21 Fifteen minutes later, we added 40 µL of anti-A antibody or anti-B antibody to the system (the same antibodies described in the section "Blood samples").…”

Section: Qd Bioconjugationmentioning

confidence: 99%

Blood group antigen studies using CdTe quantum dots and flow cytometry

Filho¹,

Pereira²,

Fernandes³

et al. 2015

IJN

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New methods of analysis involving semiconductor nanocrystals (quantum dots [QDs]) as fluorescent probes have been highlighted in life science. QDs present some advantages when compared to organic dyes, such as size-tunable emission spectra, broad absorption bands, and principally exceptional resistance to photobleaching. Methods applying QDs can be simple, not laborious, and can present high sensibility, allowing biomolecule identification and quantification with high specificity. In this context, the aim of this work was to apply dual-color CdTe QDs to quantify red blood cell (RBC) antigen expression on cell surface by flow cytometric analysis. QDs were conjugated to anti-A or anti-B monoclonal antibodies, as well as to the anti-H ( Ulex europaeus I) lectin, to investigate RBCs of A 1 , B, A 1 B, O, A 2 , and A weak donors. Bioconjugates were capable of distinguishing the different expressions of RBC antigens, both by labeling efficiency and by flow cytometry histogram profile. Furthermore, results showed that RBCs from A weak donors present fewer amounts of A antigens and higher amounts of H, when compared to A 1 RBCs. In the A group, the amount of A antigens decreased as A 1 > A 3 > A X = A el , while H antigens were A X = A el > A 1 . Bioconjugates presented stability and remained active for at least 6 months. In conclusion, this methodology with high sensibility and specificity can be applied to study a variety of RBC antigens, and, as a quantitative tool, can help in achieving a better comprehension of the antigen expression patterns on RBC membranes.

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Section: Qd Bioconjugationmentioning

confidence: 99%

Blood group antigen studies using CdTe quantum dots and flow cytometry

Filho¹,

Pereira²,

Fernandes³

et al. 2015

IJN

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“…These were first chemically modified by the incorporation of alkyne groups into amine groups from proteins by carbodiimide chemistry [82]. These authors conjugated azide-Fluor 545 (a fluorescent compound) to activated EVs.…”

Section: Semi-synthetic Exosomes: Biotechnological Modification Of Namentioning

confidence: 99%

Therapeutic biomaterials based on extracellular vesicles: classification of bio‐engineering and mimetic preparation routes

García‐Manrique

Matos

Gutiérrez

et al. 2018

J of Extracellular Vesicle

152

153

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Extracellular vesicles (EVs) are emerging as novel theranostic tools. Limitations related to clinical uses are leading to a new research area on design and manufacture of artificial EVs. Several strategies have been reported in order to produce artificial EVs, but there has not yet been a clear criterion by which to differentiate these novel biomaterials. In this paper, we suggest for the first time a systematic classification of the terms used to build up the artificial EV landscape, based on the preparation method. This could be useful to guide the derivation to clinical trial routes and to clarify the literature. According to our classification, we have reviewed the main strategies reported to date for their preparation, including key points such as: cargo loading, surface targeting strategies, purification steps, generation of membrane fragments for the construction of biomimetic materials, preparation of synthetic membranes inspired in EV composition and subsequent surface decoration.

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“…Similarly, the modified common carbodiimide method described by Hermanson [37] was used for covalent immobilization of HRP onto carboxylate-modified Sera-Mag magnetic particles. Here, 1 mg of magnetic particles was washed five times with 0.1 M phosphate buffer (pH 7.3) and activated by 7.5 mg of EDC and 1.25 mg of sulfo-NHS dissolved in 1 ml of phosphate buffer for 10 min at room temperature with gentle mixing.…”

Section: Immobilization Of Hrp On Sera-mag Microparticlesmentioning

confidence: 99%

“…The two kinds of magnetic particles-p(GMA-MOEAA)-NH 2 coated with hyaluronic acid (HA) [36] and commercially available carboxylate-modified Sera-Mag magnetic particles-were exploited for covalent coupling of specific antibodies using a slightly modified two-step carbodiimide method and with EDC as zero-length cross-linker and sulfo-NHS according to Hermanson [37]. Here, 1 mg of magnetic particles was washed five times with 50 mM Mes buffer (pH 5.0).…”

Section: Immobilization Of Anti-ova Antibodies On Carboxylate-modifiementioning

confidence: 99%