kilograms  (Seca)

Journal: Organic Process Research & Development

Article Title: Scalable Membrane Enabled One-Pot Liquid-Phase Oligonucleotide Synthesis

doi: 10.1021/acs.oprd.5c00117

Figure Lengend Snippet: Schematic representation of scalable one-pot liquid-phase oligonucleotide synthesis (OP-LPOS) enhanced by organic solvent resistant (OSR) ultrafiltration/nanofiltration membrane separation.

Article Snippet: To compare to other LPOS strategies, we evaluated two methods: the kilogram scale convergent LPOS precipitation approach by Zhou et al. at Biogen, which had a PMI ∼ 63 kg/kg, and the membrane-based nanostar sieving approach by Kim et al. from the Livingston group, which had a PMI ∼ 2480 kg/kg., , , For our developed OP-LPOS via UF membrane process, the PMI was estimated for producing 1 kg of a 20-mer PS 2′-OMe oligonucleotide based upon average values obtained experimentally (filtration yield of 99.6%, coupling efficiency of 98%, filtration and reaction conc. of 1% w/v and 12% w/v respectively, 5 diavolumes to purity and 2 diavolumes to collect) and input into the ACS Green Chemistry PMI calculator.

Techniques: Oligonucleotide Synthesis, Solvent, Membrane

Journal: Organic Process Research & Development

Article Title: Scalable Membrane Enabled One-Pot Liquid-Phase Oligonucleotide Synthesis

doi: 10.1021/acs.oprd.5c00117

Figure Lengend Snippet: Schematic representation of the overall process developed. One-pot LPOS chain extension conditions per arm: (i) Coupling: 2’-OMe phosphoramidite (1.5 equiv), DCI (4 equiv), 10% w/v 3 Å molecular sieves, MeCN, RT, 30–60 min; amidite quench: MeOH (5 equiv); (ii) sulfurization: PADS (9 equiv) or POS (3 equiv), MeCN, RT, 60 min; (iii) detritylation: TFA (15–180 equiv) and CySH (5–15 equiv), MeCN:DCM (2:1 v/v), 0 °C, 30–130 min; acid quench: pyridine (20–185 equiv), MeCN:DCM (2:1 v/v), 0 °C, 10 min; (iv) diafiltration purification using ceramic OSN/OSU membrane. Cycle is repeated n times until desired sequence is reached. [Me: methyl, Dmtr: dimethoxytrityl, ib: isobutyryl, bz: benzoyl, ac: acetyl, Cne: cyanoethyl, iPr: isopropyl].

Article Snippet: To compare to other LPOS strategies, we evaluated two methods: the kilogram scale convergent LPOS precipitation approach by Zhou et al. at Biogen, which had a PMI ∼ 63 kg/kg, and the membrane-based nanostar sieving approach by Kim et al. from the Livingston group, which had a PMI ∼ 2480 kg/kg., , , For our developed OP-LPOS via UF membrane process, the PMI was estimated for producing 1 kg of a 20-mer PS 2′-OMe oligonucleotide based upon average values obtained experimentally (filtration yield of 99.6%, coupling efficiency of 98%, filtration and reaction conc. of 1% w/v and 12% w/v respectively, 5 diavolumes to purity and 2 diavolumes to collect) and input into the ACS Green Chemistry PMI calculator.

Techniques: Diafiltration Assay, Purification, Membrane, Sequencing

Journal: Organic Process Research & Development

Article Title: Scalable Membrane Enabled One-Pot Liquid-Phase Oligonucleotide Synthesis

doi: 10.1021/acs.oprd.5c00117

Figure Lengend Snippet: HPLC reaction and diafiltration monitoring using an OP-LPOS and UF 2000 Da membrane from 20 kDa 4-arm monomer PEG-mU–OH 3c to purified dimer PEG-mUmU–OH 3d . (Blue) HPLC trace for the starting detritylated PEG-bound uridine. Post coupling-sulfurization reaction; phosphoramidite related species are visible between 16 and 19 min, POS eluted at 13.5 min, DCI eluted at 2 min and the PEG-dimer-ODmtr peaks came out at around 20 min.(Blue) After detritylation and acid quench (feed); a peak for Dmtr-scavenger is visible at 25 min with detritylated amidites species eluting between 0 and 15 min, and the detritylated PEG-dimer–OH peak appearing at 18 min. Permeate and retentate HPLC traces are also shown where final PEG-dimer–OH 3c showed 99.5% rejection by membrane and 98.0% Area purity by HPLC.

Article Snippet: To compare to other LPOS strategies, we evaluated two methods: the kilogram scale convergent LPOS precipitation approach by Zhou et al. at Biogen, which had a PMI ∼ 63 kg/kg, and the membrane-based nanostar sieving approach by Kim et al. from the Livingston group, which had a PMI ∼ 2480 kg/kg., , , For our developed OP-LPOS via UF membrane process, the PMI was estimated for producing 1 kg of a 20-mer PS 2′-OMe oligonucleotide based upon average values obtained experimentally (filtration yield of 99.6%, coupling efficiency of 98%, filtration and reaction conc. of 1% w/v and 12% w/v respectively, 5 diavolumes to purity and 2 diavolumes to collect) and input into the ACS Green Chemistry PMI calculator.

Techniques: Diafiltration Assay, Membrane, Purification

Journal: Organic Process Research & Development

Article Title: Scalable Membrane Enabled One-Pot Liquid-Phase Oligonucleotide Synthesis

doi: 10.1021/acs.oprd.5c00117

Figure Lengend Snippet: Comparison of PMI per nucleotide for the developed OP-LPOS process enhanced by OSU membrane filtration compared to the current LPOS and SPOS methods.

Article Snippet: To compare to other LPOS strategies, we evaluated two methods: the kilogram scale convergent LPOS precipitation approach by Zhou et al. at Biogen, which had a PMI ∼ 63 kg/kg, and the membrane-based nanostar sieving approach by Kim et al. from the Livingston group, which had a PMI ∼ 2480 kg/kg., , , For our developed OP-LPOS via UF membrane process, the PMI was estimated for producing 1 kg of a 20-mer PS 2′-OMe oligonucleotide based upon average values obtained experimentally (filtration yield of 99.6%, coupling efficiency of 98%, filtration and reaction conc. of 1% w/v and 12% w/v respectively, 5 diavolumes to purity and 2 diavolumes to collect) and input into the ACS Green Chemistry PMI calculator.

Techniques: Comparison, Membrane, Filtration

Journal: Animals : an Open Access Journal from MDPI

Article Title: Effect of Dietary Benzoic Acid and Oregano Essential Oil as a Substitute for an Anti-Coccidial Agent on Growth Performance and Physiological and Immunological Responses in Broiler Chickens Challenged with Eimeria Species

doi: 10.3390/ani14203008

Figure Lengend Snippet: Description of the experimental design.

Article Snippet: Ltd., Ahmedabad, India) at 500 g/kg to obtain an effective dose of 60 mg salinomycin per kilogram in the finished feed [ ] and coccidia -challenged (Sal); (iv) supplemented with BA (Hi-LR TM , Himedia Laboratories Pvt.

Techniques:

Journal: Animals : an Open Access Journal from MDPI

Article Title: Effect of Dietary Benzoic Acid and Oregano Essential Oil as a Substitute for an Anti-Coccidial Agent on Growth Performance and Physiological and Immunological Responses in Broiler Chickens Challenged with Eimeria Species

doi: 10.3390/ani14203008

Figure Lengend Snippet: Optical density (at 492 nm) of the antibody titer level in diluted serum of broiler chickens supplemented with different anticoccidial agents and challenged with coccidia. NC = negative control chickens, PC = positive control chickens, coccidia-challenged and non-treated, Sal = coccidia-challenged chickens + salinomycin at 60 mg/kg of feed, BA = coccidia-challenged chickens + benzoic acid at 500 mg/kg of feed, OEO = coccidia-challenged + oregano essential oil at 500 mg/kg of feed, B&O = coccidia-challenged + benzoic acid at 500 mg/kg of feed + oregano essential oil at 500 mg/kg of feed, Trt = treatment, DPI = days post-infection. a,b,c,d,e Means with different letters for each column bar differ significantly ( p < 0.05) among the treatments (Fisher’s protected least significant difference test).

Article Snippet: Ltd., Ahmedabad, India) at 500 g/kg to obtain an effective dose of 60 mg salinomycin per kilogram in the finished feed [ ] and coccidia -challenged (Sal); (iv) supplemented with BA (Hi-LR TM , Himedia Laboratories Pvt.

Techniques: Negative Control, Positive Control, Infection

Journal: Animals : an Open Access Journal from MDPI

Article Title: Effect of Dietary Benzoic Acid and Oregano Essential Oil as a Substitute for an Anti-Coccidial Agent on Growth Performance and Physiological and Immunological Responses in Broiler Chickens Challenged with Eimeria Species

doi: 10.3390/ani14203008

Figure Lengend Snippet: Relative expression (fold change) of ( A ) interleukin 10, ( B ) interferon gamma, and ( C ) Toll-like receptor 4 genes in the jejunum of broiler chickens supplemented with different anticoccidial agents and challenged with coccidia. NC = negative control chickens, PC = positive control chickens, coccidia-challenged and non-treated, Sal = coccidia-challenged chickens + salinomycin at 60 mg/kg of feed, BA = coccidia-challenged chickens + benzoic acid at 500 mg/kg of feed, OEO = coccidia-challenged + oregano essential oil at 500 mg/kg of feed, B&O = coccidia-challenged + benzoic acid at 500 mg/kg of feed + oregano essential oil at 500 mg/kg of feed, DPI = days post-infection. a,b,c,d,e Means with different letters for each column bar differ significantly ( p < 0.05) among the treatments (Fisher’s protected least significant difference test).

Article Snippet: Ltd., Ahmedabad, India) at 500 g/kg to obtain an effective dose of 60 mg salinomycin per kilogram in the finished feed [ ] and coccidia -challenged (Sal); (iv) supplemented with BA (Hi-LR TM , Himedia Laboratories Pvt.

Techniques: Expressing, Negative Control, Positive Control, Infection