Scalable One-Pot - Liquid-Phase Oligonucleotide Synthesis for Model Network Hydrogels

2020 ◽  
Author(s):  
Guido Creusen ◽  
Cecilia Oluwadunsin Akintayo ◽  
Katja Schumann ◽  
Andreas Walther
Keyword(s):  
Liquid Phase ◽  
Solid Phase ◽  
Adenine Nucleotides ◽  
Building Blocks ◽  
Oligonucleotide Synthesis ◽  
One Pot ◽  
Model Network ◽  
Dna And Rna ◽  
Sequence Recognition ◽  
Sequential Coupling

Solid-phase oligonucleotide synthesis (SPOS) based on phosphoramidite chemistry is currently the most widespread technique for DNA and RNA synthesis, but suffers from scalability limitations and high reagent consumption. Liquid-phase oligonucleotide synthesis (LPOS) uses soluble polymer supports and has the potential of being scalable. However, at present, LPOS requires 3 separate reaction steps and 4-5 precipitation steps per nucleotide addition. Moreover, long acid exposure times during the deprotection step degrade sequences with high A-content (adenine) due to depurination and chain cleavage. In this work, we present the first one-pot liquid-phase DNA synthesis technique, which allows the addition of one nucleotide in a one-pot reaction of sequential coupling, oxidation and deprotection, followed by a single precipitation step. Furthermore, we demonstrate how to suppress depurination during the addition of adenine nucleotides. We showcase the potential of this technique to prepare high-purity 4-arm PEG‑T<sub>20</sub> (T = thymine) and 4-arm PEG-A<sub>20</sub>building blocks in multi-gram scale. Such complementary 4-arm PEG-DNA building blocks reversibly self-assemble into supramolecular model network hydrogels, and facilitate the elucidation of bond lifetimes. These model network hydrogels exhibit new levels of mechanical properties, high stability at room temperature (melting at 44 ‎°C), and thus open up pathways to next-generation, scalable DNA-materials programmable through sequence recognition and available for macroscale applications<i>.</i>

scholarly journals Scalable One-Pot - Liquid-Phase Oligonucleotide Synthesis for Model Network Hydrogels

2020 ◽  
Author(s):  
Guido Creusen ◽  
Cecilia Oluwadunsin Akintayo ◽  
Katja Schumann ◽  
Andreas Walther
Keyword(s):  
Liquid Phase ◽  
Solid Phase ◽  
Adenine Nucleotides ◽  
Building Blocks ◽  
Oligonucleotide Synthesis ◽  
One Pot ◽  
Model Network ◽  
Dna And Rna ◽  
Sequence Recognition ◽  
Sequential Coupling

Solid-phase oligonucleotide synthesis (SPOS) based on phosphoramidite chemistry is currently the most widespread technique for DNA and RNA synthesis, but suffers from scalability limitations and high reagent consumption. Liquid-phase oligonucleotide synthesis (LPOS) uses soluble polymer supports and has the potential of being scalable. However, at present, LPOS requires 3 separate reaction steps and 4-5 precipitation steps per nucleotide addition. Moreover, long acid exposure times during the deprotection step degrade sequences with high A-content (adenine) due to depurination and chain cleavage. In this work, we present the first one-pot liquid-phase DNA synthesis technique, which allows the addition of one nucleotide in a one-pot reaction of sequential coupling, oxidation and deprotection, followed by a single precipitation step. Furthermore, we demonstrate how to suppress depurination during the addition of adenine nucleotides. We showcase the potential of this technique to prepare high-purity 4-arm PEG‑T<sub>20</sub> (T = thymine) and 4-arm PEG-A<sub>20</sub>building blocks in multi-gram scale. Such complementary 4-arm PEG-DNA building blocks reversibly self-assemble into supramolecular model network hydrogels, and facilitate the elucidation of bond lifetimes. These model network hydrogels exhibit new levels of mechanical properties, high stability at room temperature (melting at 44 ‎°C), and thus open up pathways to next-generation, scalable DNA-materials programmable through sequence recognition and available for macroscale applications<i>.</i>

scholarly journals Scalable One-Pot-Liquid-Phase Oligonucleotide Synthesis for Model Network Hydrogels

2020 ◽  
Vol 142 (39) ◽  
pp. 16610-16621
Author(s):  
Guido Creusen ◽  
Cecilia Oluwadunsin Akintayo ◽  
Katja Schumann ◽  
Andreas Walther
Keyword(s):  
Liquid Phase ◽  
Oligonucleotide Synthesis ◽  
One Pot ◽  
Model Network

Fully Automated Parallel Oligonucleotide Synthesizer

2001 ◽  
Vol 66 (8) ◽  
pp. 1299-1314 ◽  
Author(s):  
Michal Lebl ◽  
Christine Burger ◽  
Brett Ellman ◽  
David Heiner ◽  
Georges Ibrahim ◽  
...  
Keyword(s):  
Solid Phase ◽  
Building Blocks ◽  
Solid Support ◽  
Continuous Operation ◽  
Gear Pump ◽  
Oligonucleotide Synthesis ◽  
Center Of Rotation ◽  
Microtiter Plates ◽  
The Individual ◽  
Design And Construction

Design and construction of automated synthesizers using the tilted plate centrifugation technology is described. Wash solutions and reagents common to all synthesized species are delivered automatically through a 96-channel distributor connected to a gear pump through two four-port selector valves. Building blocks and other specific reagents are delivered automatically through banks of solenoid valves, positioned over the individual wells of the microtiterplate. These instruments have the following capabilities: Parallel solid-phase oligonucleotide synthesis in the wells of polypropylene microtiter plates, which are slightly tilted down towards the center of rotation, thus generating a pocket in each well, in which the solid support is collected during centrifugation, while the liquid is expelled from the wells. Eight microtiterplates are processed simultaneously, providing thus a synthesizer with a capacity of 768 parallel syntheses. The instruments are capable of unattended continuous operation, providing thus a capacity of over two millions 20-mer oligonucleotides in a year.

A Comprehensive One-Pot Synthesis of Protected Cysteine and Selenocysteine SPPS Derivatives

2014 ◽  
Vol 21 (12) ◽  
pp. 1257-1264
Author(s):  
Stevenson Flemer
Keyword(s):  
Solid Phase ◽  
Solid Phase Peptide Synthesis ◽  
Reduction Process ◽  
Building Blocks ◽  
High Yield ◽  
One Pot ◽  
Peptide Models ◽  
Direct Use ◽  
Proof Of Principle ◽  
Subsequent Incorporation

A proof-of-principle methodology is presented in which all commercially-available cysteine (Cys) and selenocysteine (Sec) solid phase peptide synthesis (SPPS) derivatives are synthesized in high yield from easily prepared protected dichalcogenide precursors. A Zn-mediated biphasic reduction process applied to a series of four bis-Nα-protected dichalcogenide compounds allows facile conversion to their corresponding thiol and selenol intermediates followed by insitu S- or Se-alkylation with various electrophiles to directly access twenty one known Cys and Sec SPPS derivatives. Most of these derivatives were able to be precipitated in crude form out of petroleum ether in sufficient purity for direct use as peptide building blocks. Subsequent incorporation of these derivatives into peptide models nicely illustrates their viability and applicability toward SPPS.

Identification of hepatitis a virus in cell cultures by solid phase immune electron microscopy

1986 ◽  
Vol 44 ◽  
pp. 238-239
Author(s):  
C.D. Humphrey ◽  
T.L. Cromeans ◽  
E.H. Cook ◽  
D.W. Bradley
Keyword(s):  
Electron Microscopy ◽  
Liquid Phase ◽  
Cell Cultures ◽  
Rapid Detection ◽  
Hepatitis A ◽  
Hepatitis A Virus ◽  
Solid Phase ◽  
Immune Electron Microscopy ◽  
Detection And Identification

There is a variety of methods available for the rapid detection and identification of viruses by electron microscopy as described in several reviews. The predominant techniques are classified as direct electron microscopy (DEM), immune electron microscopy (IEM), liquid phase immune electron microscopy (LPIEM) and solid phase immune electron microscopy (SPIEM). Each technique has inherent strengths and weaknesses. However, in recent years, the most progress for identifying viruses has been realized by the utilization of SPIEM.

SYNTHESIS OF HYDROGEN IN ACOUSTOPLASMA DISCHARGE IN A LIQUID-PHASE STREAM

2019 ◽  
pp. 42-48 ◽  
Author(s):  
N. A. Bulychev
Keyword(s):  
Liquid Phase ◽  
Organic Compounds ◽  
Electrode Materials ◽  
Solid Phase ◽  
Plasma Discharge ◽  
Raw Material ◽  
Two Phase ◽  
Reduced Pressure ◽  
External Power Source ◽  
Phase Medium

In this paper, the plasma discharge in a high-pressure fluid stream in order to produce gaseous hydrogen was studied. Methods and equipment have been developed for the excitation of a plasma discharge in a stream of liquid medium. The fluid flow under excessive pressure is directed to a hydrodynamic emitter located at the reactor inlet where a supersonic two-phase vapor-liquid flow under reduced pressure is formed in the liquid due to the pressure drop and decrease in the flow enthalpy. Electrodes are located in the reactor where an electric field is created using an external power source (the strength of the field exceeds the breakdown threshold of this two-phase medium) leading to theinitiation of a low-temperature glow quasi-stationary plasma discharge.A theoretical estimation of the parameters of this type of discharge has been carried out. It is shown that the lowtemperature plasma initiated under the flow conditions of a liquid-phase medium in the discharge gap between the electrodes can effectively decompose the hydrogen-containing molecules of organic compounds in a liquid with the formation of gaseous products where the content of hydrogen is more than 90%. In the process simulation, theoretical calculations of the voltage and discharge current were also made which are in good agreement with the experimental data. The reaction unit used in the experiments was of a volume of 50 ml and reaction capacity appeared to be about 1.5 liters of hydrogen per minute when using a mixture of oxygen-containing organic compounds as a raw material. During their decomposition in plasma, solid-phase products are also formed in insignificant amounts: carbon nanoparticles and oxide nanoparticles of discharge electrode materials.

Synthesis of Nitrile-Bearing Quaternary Centers via an Equilibrium-Driven Transnitrilation and Anion-Relay Strategy

2019 ◽  
Author(s):  
Sebastien Alazet ◽  
Michael West ◽  
Purvish Patel ◽  
Sophie Rousseaux
Keyword(s):  
Building Blocks ◽  
One Pot ◽  
Quaternary Centers ◽  
Synthetic Intermediates

The efficient preparation of nitrile-containing building blocks is of interest due to their utility as synthetic intermediates and their prevalence in pharmaceuticals. As a result, significant efforts have been made to develop methods to access these motifs which rely on safer and non-toxic sources of CN. Herein, we report that 2-methyl-2-phenylpropanenitrile is an efficient, non-toxic, electrophilic CN source for the synthesis of nitrile-bearing quaternary centers via a thermodynamic transnitrilation and anion-relay strategy. This one-pot process leads to nitrile products resulting from the gem-difunctionalization of alkyl lithium reagents.<br>

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