URui Huang, bo xu
Isikhungo se-R & D Center
Ukuqalisa
I-peptide iyindawo ehlanganisiwe ehlanganiswa ama-amino acid, ngalinye lawo elinezici ezihlukile zomzimba nangamakhemikhali ngenxa yezinhlobo ezahlukahlukene nokuhleleka kwezinsalela ze-amino acid kwakha ukulandelana kwayo. Ngokuthuthukiswa kwe-syntenisis yesigaba esiqinile, ukuhlanganiswa kwamakhemikhali kwama-peptides ahlukahlukene asebenzayo kuye kwathuthuka kakhulu. Kodwa-ke, ngenxa yokwakheka okuyinkimbinkimbi kwe-peptide etholwe ngokuhlanganiswa kwesigaba esiqinile, umkhiqizo wokugcina kufanele uhlanziwe ngezindlela ezithembekile zokuhlukanisa. Izindlela zokuhlanza ezivame ukusetshenziswa ze-pottides zifaka i-ion Exchange Chromatography (IEC) kanye ne-Rp-HORDER-SPETIWECH CRROMOTIDES (MW-HPLC), i-Expression imishini yokuhlukanisa, i-application yokwehlukaniswa okusheshayo, icala eliphumelelayo lalishicilelwe ngaphambilini icala lakwaSantai, Lapho kusetshenziswa i-SepaFlash Rp C18 Cartridge ngokuhlanzwa okusheshayo kweThymopentin (TP-5) kanye nomhlangano womkhiqizo oqondiwe izidingo ezitholakele.
Umdwebo 1. Ama-amino acid ajwayelekile (akhiqizwa kabusha kusuka ku-www.Bachem.com).
Kunezinhlobo ezingama-20 zama-amino acid ajwayelekile ekwakhiweni kwama-peptides. Lawa ma-amino acid angahlukaniswa amaqembu alandelayo ngokuya ngePolarity ne-Acid-Base Property: I-Non-Polar (Hydrophobic), i-Polar (engangenisiwe), i-acidic noma eyisisekelo (njengoba kukhonjisiwe kuMfanekiso 1). Ekulandekelweni kwe-peptide, uma ama-amino acid akha ukulandelana ikakhulukazi amaphalane (njengoba amakwe ngombala opinki kuMdwebo 1), njenge-cysteinine, i-areatine, i-threinine, i-thylide, njll. Ngesikhathi senqubo yokuhlanza yalezi zinhlaka eziqinile ze-polar peptic nge-chromatography ebuyiselwe emuva, into ebizwa ngokuthi yi-hydromobic yesigaba ubuchwepheshe izokwenzeka (bhekana nenothi lesicelo elishicilelwe ngaphambilini: I-Hydrophobic Technology Collappy: I-AQ ibuyiselwe amakholomu we-ChromaTography we-ChromaTography kanye nezicelo zabo). Uma kuqhathaniswa namakholomu ajwayelekile e-C18, amakholomu we-C18AQ athuthukisiwe alungele ukuhlanzwa kwamasampula aqinile we-polar noma ama-hydrophilic. Kulokhu okuthunyelwe, kwasetshenziswa i-polar peptide eqinile njengesampula futhi ihlanjululwe yikholomu ye-C18AQ. Ngenxa yalokhu, umhlangano womkhiqizo oqondiwe izidingo ezitholakele futhi zingasetshenziswa kulolo cwaningo olulandelayo.
| Into yokwenza utho | I-Sepabean™umshini 2 | |||
| Ama-cartridges | I-12 g sepaflash C18 RP Flash Carthridge (Spherical Silica, 20 - 45 μm, i-100 Å, Order Nanber: SW-5222-012-SP) | I-12 g sepaflash C18AQ RP Flash Carthridge (spherical Silica, 20 - 45 μm, Inombolo ye-Order: Inombolo ye-SW-5222-012-SP (AQ)) | ||
| Umbuzi | 254 NM, 220 NM | 214 NM | ||
| Isigaba | Solvent a: amanzi Solvent b: acetonitrile | |||
| Izinga Lokugeleza | 15 ml / min | 20 ml / min | ||
| Ukulayisha isampula | 30 mg | |||
| I-gradient | Isikhathi (CV) | Solvent b (%) | Isikhathi (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 | |||
Imiphumela nengxoxo
Ukuqhathanisa ukusebenza kokuhlanzeka kwesampula ye-polar peptide phakathi kwekholomu ejwayelekile ye-C18 kanye nekholamu ye-C18AQ, sisebenzise ikholomu ejwayelekile ye-C18 yokuhlanzwa kwe-flash kwesampula njengokuqala kwesampula njengokuqala kwesampula njengokuqala. Njengoba kukhonjisiwe kuMdwebo 2, ngenxa yokuwohloka kwesigaba se-hydrophobic kwamaketanga we-C18 adalwe yisilinganiso esiphakeme samanzi, isampula lagcinwa kancane ku-C18 cartridge elijwayelekile futhi lalivunyiswa ngokuqondile ngesigaba seselula. Ngenxa yalokho, isampula lalingahlukaniswa ngempumelelo futhi lihlanzwa.
Umdwebo 2. I-flash chromatograph yesampula ku-C18 Cartridge.
Okulandelayo, sasebenzisa ikholomu ye-C18AQ yokuhlanzwa kwe-Flash kwesampula. Njengoba kukhonjisiwe kuMdwebo 3, i-peptide yagcinwa ngempumelelo kwikholamu bese ichayeka. Umkhiqizo oqondiwe wahlukaniswa nokungcola kwisampula eluhlaza futhi eqoqwe. Ngemuva kwe-lyophilization bese kuhlaziywa yi-HPLC, umkhiqizo ohlanziwe unobumsulwa bama-98.2% futhi bungasetshenziswa futhi ukuze kusetshenziswe ucwaningo olulandelayo lwe-State Research and Development.
Umdwebo 3. I-flash chromatograph yesampula ku-C18AQ cartridge.
Ekuphetheni, Sepaflash C18AQ RP Flash Carthridy ehlanganiswe ne-Flash Chromatography Syste Sepabean™Umshini unganikeza isixazululo esisheshayo nesisebenzayo sokuhlanzwa kwamasampula aqinile we-polar noma amasampula e-hydrophilic.
Kunochungechunge lwama-cartridgeges we-sepaflash C18AQ RP Flash anokucaciswa okuhlukile kubuchwepheshe beSantai (njengoba kukhonjisiwe kuThebula 2).
| Inombolo yento | Usayizi wekholomu | Izinga Lokugeleza (ml / min) | Max.prestrure (PSI / Bar) |
| 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 |
Ithebula 2. Sepaflash C18AQ RP Flash Cartridges. Izinto zokupakisha: I-Spherical C18 esebenza kahle kakhulu
Ukuthola eminye imininingwane ngokuchazwa okunemininingwane yomshini we-Sepabean ™, noma imininingwane yoku-oda kuma-cartridge we-Sepaflash Series, sicela uvakashele iwebhusayithi yethu.
Isikhathi sePosi: Oct-12-2018