Hits:6674 , posted at 2020/4/14 15:31
An
oligonucleotide synthesizer is an automated instrument designed to synthesize oligonucleotides
structurally similar to Oligo DNA/RNA Synthesizer.
CSBio Oligo DNA/RNA Synthesizer release provides a better choice for
domestic researchers who are developing new oligonucleotide drugs.(that can be used as gene therapies for some rare
diseases and diseases caused by viruses)
The following
is the CSBio Oligo DNA/RNA Synthesizer advantages and detailed introduction:
Utilization of
Polystyrene
Why Polystyrene?
•
More oligo therapeutics are approaching commercialization
•
Reduction in production costs become a
critical consideration
•
Increasing solid support loading and managing synthesis
efficiencycan greatly control costs
•
CPG will typically be in 0.06 mmol/g, compared to
polystyrene in the0.3 mmol/g range.5x increase in loading
•
Scale up is challenging
•
Traditional column loading needs to
manage polystyrene swelling -limiting the total resin loaded, adding column
dead volume for resin growth,and address backpressure and channel formation
challenges.
Key System Operation
•
Mixing
is performed by 180ºinversion with argon bubbling
either from the top or bottom of the reactionvessel.
•
The reaction vessel is anopen reactor made of borosilicate glass
•
Flow
through utilized fordetritylation, inversion mixing is possible during flow
through.
•
UV
detection fordetritylation
•
Argon
line/reservoir purgingutilized to maintain inert atmosphere
•
8x
individual amiditepositions (more at request)
•
Circulation is possible withthe system
Synthesis Protocol
1.Resin swelling (acetonitrile anhydrous)
2. Resin washing (acetonitrile anhydrous)
3. De-blocking (3% TCA in Toluene)
4. Resin washing (acetonitrile anhydrous)
5. Amidite and Activator transferring/mixing into RV
6.
Coupling Activator stock solution] & Amidite stock solution
7. Resin washing (acetonitrile anhydrous)
8. Oxidation (Oxidation stock solution)
9. Resin washing (acetonitrile)
10. Capping: CapA/CapB
11. Resin washing (acetonitrile anhydrous)
12. Start from Step (2) for the next amidite
13.Final DMT deblocking same as in Step (3)
Discussion and Next Steps
•
Conventional mixing is to flow past stationary resin that is packedwithin a
column. High pressure occurs as a result due to the need to flow athigh speeds
relative to column size to achieve mixing. CSBio’s
synthesizermixing is performed by suspending the resin in the reaction
solution, allowingfor the resin to achieve high reaction and washing
efficiency.
•
VIMAD
and Nitto resin provided similar results
•
Quality of ETT made a big difference – a higher quality
supplier forETT gave a 3-5% higher HPLC purity for the 5’-ACGT-3’ synthesis
•
Further protocol evaluation with different coupling and deblockmethods,
improved raw material handling and preparation, and longer oligos
•
Evaluation of CPG resin with new synthesis reactor
•
Development of a resin specific for this type of reactor design canpotentially
yield significantly better synthesis results. All resins usedto-date have been
optimized for column based reactors.
CSBio Introduction
CSBio is Global peptide company bringing more than 26 years of technical expertise into the peptide industry which cover three main categories of products and services:
1) peptide CDMO & CMO service -GMP and R&D peptide production, with API peptide production approved by USFDA.
2) Peptide Instrumentation - from R&D up to Large Scale Manufacturing-type synthesizer, Cleavage systems, PreHPLC system, Column packing equipment and peptide production technical support.
3) Peptide raw material -- High Quality Standard Grade and High Grade Fmoc-protected Amino Acid, peptide reagents, coupling reagents, resins and special reagents, supported by relevant validated QC data, stability data and DMF technical documents.
4) DNA/RNA oligonucleotide synthesizers that can be used from research、process development and production.
CSBio (Shanghai) Ltd.
China (Shanghai) Pilot Free Trade Zone, 85 Faladi Road, Building 2,Room 502 ,Shanghai 201210, China
Email:info@csbiochina.com
Tel:+8621-50781653/50781655
Fax:+8621-50781652/021-50781656
Website: www.csbiochina.com www.csbio.com
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