Rui Huang, Bo Xu
Talosaga R&D Nofoaga Autu
Folasaga
O le peptide o se tuufaatasiga e aofia ai amino acids, o ia mea taʻitasi e iai uiga faʻapitoa ma vailaʻau faʻapitoa ona o ituaiga eseese ma le faʻasologa o toega amino acid e aofia ai lona faʻasologa.Faatasi ai ma le atinaʻeina o le faʻaogaina o vailaʻau faʻasolosolo, o le faʻaogaina o vailaʻau o peptides malosi eseese ua matua alualu i luma.Ae ui i lea, ona o le faʻalavelave lavelave o le peptide e maua mai i le faʻasologa o vaega faʻamalosi, o le oloa mulimuli e tatau ona faʻamamaina e ala i auala faʻalagolago.O auala masani faʻamamaina mo peptides e aofia ai chromatography fesuiaiga o ion (IEC) ma faʻaliliuga-vaega maualuga chromatography vai (RP-HPLC), o loʻo i ai le faʻaletonu o le maualalo o le faʻataʻitaʻiga faʻataʻitaʻiga gafatia, tau maualuga o le vavaeeseina o ala o faasalalauga, lavelave ma taugata meafaigaluega vavaeeseina, ma isi. Mo le vave faʻamamaina o peptides mole laiti (MW <1 kDa), na faʻasalalau muamua se mataupu talosaga manuia e Santai Technologies, lea na faʻaaogaina ai se pusa SepaFlash RP C18 mo le faʻamamaina vave o le thymopentin (TP-5) ma le oloa fa'atatau e fetaui ma mana'oga na maua.
Ata 1. 20 amino acids masani (toe gaosia mai www.bachem.com).
E 20 ituaiga o amino acids e taatele i le tuufaatasiga o peptides.O nei amino acids e mafai ona vaevaeina i vaega nei e tusa ai ma lo latou polarity ma le acid-base meatotino: non-polar (hydrophobic), polar (leai totogi), acidic poʻo le faavae (e pei ona faʻaalia i le Ata 1).I se peptide sequence, afai o amino acids e aofia ai le faasologa e tele lava polar (e pei ona faailogaina i le lanu piniki i le Ata 1), e pei o Cysteine, Glutamine, Asparagine, Serine, Threonine, Tyrosine, ma isi, o lenei peptide atonu e malosi. polarity ma e matua solu i le vai.I le taimi o le fa'amamāina o nei fa'ata'ita'iga polar peptide malosi e ala i le chromatography fa'aliliuina, o le a tupu se mea e ta'ua o le hydrophobic phase collapse (silasila i le tusi talosaga na fa'asalalau muamua e Santai Technologies: Hydrophobic Phase Collapse, AQ Reversed Phase Chromatography Columns ma a latou Talosaga).Pe a faʻatusatusa i koluma masani C18, o le faʻaleleia o koluma C18AQ e sili ona fetaui mo le faʻamamaina o faʻataʻitaʻiga polar poʻo hydrophilic malosi.I lenei pou, o se peptide polar malosi na faʻaaogaina e fai ma faʻataʻitaʻiga ma faʻamamaina e se koluma C18AQ.O le i'uga, o le oloa fa'atatau e fetaui ma mana'oga na maua ma mafai ona fa'aoga i su'esu'ega ma atina'e o lo'o mulimuli mai.
| Meafaigaluega | SepaPi™masini 2 | |||
| Cartridges | 12 g SepaFlash C18 RP kesi moli (silika lapotopoto, 20 - 45 μm, 100 Å, Poloaiga nunber：SW-5222-012-SP) | 12 g SepaFlash C18AQ RP kesi moli (silika lapotopoto, 20 - 45 μm, 100 Å, Numera oka:SW-5222-012-SP(AQ)) | ||
| Umi ole galu | 254 nm, 220 nm | 214 nm | ||
| Vaega feavea'i | Solvent A: Vai Solvent B: Acetonitrile | |||
| Fuafuaga tafe | 15 mL/min | 20 mL/min | ||
| Fa'ata'ita'iga uta | 30 mg | |||
| Gradient | Taimi (CV) | Solvent B (%) | Taimi (min) | Solvent B (%) |
| 0 | 0 | 0 | 4 | |
| 1.0 | 0 | 1.0 | 4 | |
| 10.0 | 6 | 7.5 | 18 | |
| 12.5 | 6 | 13.0 | 18 | |
| 16.5 | 10 | 14.0 | 22 | |
| 19.0 | 41 | 15.5 | 22 | |
| 21.0 | 41 | 18.0 | 38 | |
| / | / | 20.0 | 38 | |
| 22.0 | 87 | |||
| 29.0 | 87 | |||
I'uga ma Talanoaga
Ina ia faʻatusatusa le faʻatinoga o le faʻamamaina mo le faʻataʻitaʻiga o le peptide polar i le va o koluma masani C18 ma le koluma C18AQ, matou te faʻaaogaina se koluma masani C18 mo le faʻamamaina moli o le faʻataʻitaʻiga e fai ma amataga.E pei ona fa'aalia i le Ata 2, ona o le pa'u o le vaega o le hydrophobic o filifili C18 e mafua mai i le maualuga o le vai, o le fa'ata'ita'iga na tau le taofia i luga o le kalama masani C18 ma sa fa'asa'o sa'o i fafo e le vaega feavea'i.O se taunuuga, o le faʻataʻitaʻiga e leʻi lelei ona vavaeeseina ma faʻamamaina.
Ata 2. O le chromatogram moli o le fa'ata'ita'iga i luga o se kesi masani C18.
O le isi, matou faʻaaogaina se koluma C18AQ mo le faʻamamaina o le faʻataʻitaʻiga.E pei ona faʻaalia i le Ata 3, o le peptide na faʻatumauina lelei i luga o le koluma ona faʻaumatia ai lea.O le oloa faʻatatau na tuʻueseese mai mea leaga i le faʻataʻitaʻiga mata ma aoina.A maeʻa le lyophilization ona suʻesuʻeina lea e le HPLC, o le oloa faʻamamaina e iai le mama o le 98.2% ma e mafai ona faʻaogaina atili mo suʻesuʻega ma atinaʻe e sosoo ai.
Ata 3. O le chromatogram moli o le fa'ata'ita'iga i luga o se kesi C18AQ.
I le fa'ai'uga, SepaFlash C18AQ RP kesi moli fa'atasi ma le fa'aogaina o le chromatography system SepaBean.™e mafai e masini ona ofoina atu se fofo vave ma aoga mo le faʻamamaina o faʻataʻitaʻiga polar poʻo hydrophilic malosi.
O loʻo i ai se faʻasologa o le SepaFlash C18AQ RP faʻapipiʻi moli ma faʻamatalaga eseese mai le Santai Technology (e pei ona faʻaalia i le Laulau 2).
| Numera o mea | Tele o le koluma | Fua Faatatau (mL/min) | Max.Omiga (psi/pa) |
| SW-5222-004-SP(AQ) | 5.4 g | 5-15 | 400/27.5 |
| SW-5222-012-SP(AQ) | 20 g | 10-25 | 400/27.5 |
| SW-5222-025-SP(AQ) | 33 g | 10-25 | 400/27.5 |
| SW-5222-040-SP(AQ) | 48 g | 15-30 | 400/27.5 |
| SW-5222-080-SP(AQ) | 105 g | 25-50 | 350/24.0 |
| SW-5222-120-SP(AQ) | 155 g | 30-60 | 300/20.7 |
| SW-5222-220-SP(AQ) | 300 g | 40-80 | 300/20.7 |
| SW-5222-330-SP(AQ) | 420 g | 40-80 | 250/17.2 |
Fuafuaga 2. SepaFlash C18AQ RP kasini moli.Mea fa'apipi'i: Silika fa'apipi'i sili C18(AQ) maualuga, 20 - 45 μm, 100 Å.
Mo nisi faʻamatalaga i faʻamatalaga auiliili o le SepaBean™ masini, poʻo le faʻatonuga o faʻamatalaga i le SepaFlash series flash cartridges, faʻamolemole asiasi i la matou upega tafaʻilagi.
Taimi meli: Oke-12-2018