Rui Huang, Bo Xu
Codsiga Xarunta R&D
Hordhac
peptide-ku waa iskudhis ka kooban asiidhyada amino, kuwaaso mid kastaa leeyahay astaamo jidheed iyo kiimiko oo gaar ah taas oo ay ugu wacan tahay noocyada iyo nidaamka kala duwan ee hadhaaga amino acid ee ka kooban isku xigxiga.Iyada oo horumarinta wejiga adag ee kiimikada kiimikada, isku-dhafka kiimikada ee peptides firfircoon ee kala duwan ayaa sameeyay horumar weyn.Si kastaba ha noqotee, iyada oo ay ugu wacan tahay isku-dhafka adag ee peptide-ka ee lagu helo isku-dhafka wajiga adag, badeecada ugu dambeysa waa in lagu nadiifiyaa habab kala-saarid la isku halleyn karo.Hababka sifaynta sida caadiga ah loo isticmaalo peptides waxaa ka mid ah chromatography sarrifka ion (IEC) iyo chromatography dareere waxqabadka sare wejiga-rogid (RP-HPLC), kuwaas oo leh faa'iido darrooyinka awoodda loading muunad hoose, qiimaha sare ee warbaahinta kala soocida, adag iyo qalab kala soocida qaali ah, iwm. Si loo sifeeyo degdega ah ee peptides molecule yaryar (MW <1 kDa), kiis arji guul leh ayaa hore u daabacay Santai Technologies, kaas oo kartridge SepaFlash RP C18 loo adeegsaday sifaynta degdega ah ee thymopentin (TP-5) iyo buuxinta shuruudaha ayaa la helay alaabta bartilmaameedka ah.
Jaantus 1. 20 amino acids caadi ah (oo laga soo saaray www.bachem.com).
Waxaa jira 20 nooc oo acids amino ah kuwaas oo ku badan halabuurka peptides.Asiidhyada amino-yada waxa loo qaybin karaa kooxahan soo socda iyadoo loo eegayo hantidooda aashitada iyo aashitada: non-polar (hydrophobic), polar (aan la qaadin), acidic ama aasaasiga ah (sida ku cad sawirka 1).Isku xigxiga peptide, haddii amino acids-ka ka kooban taxanuhu ay u badan yihiin kuwa polar (sida ku calaamadsan midabka casuuska ee sawirka 1), sida Cysteine, Glutamine, Asparagine, Serine, Threonine, Tyrosine, iwm. markaas peptide-kan waxa laga yaabaa inuu yeesho mid xooggan. polarity oo aad ugu milmi biyaha.Inta lagu guda jiro nidaamka sifaynta ee shaybaarada peptide polar xoog leh by chromatography weji rogan, ifafaale loo yaqaan hydrophobic wejiga burburka ayaa dhici doona (tixraac codsi hore loo daabacay by Santai Teknoolajiyada: Hydrophobic Wejiga Collapse, AQ Reversed Fase Chromatography Columns iyo Codsigooda).Marka la barbar dhigo tiirarka caadiga ah ee C18, tiirarka C18AQ ee la hagaajiyay ayaa aad ugu habboon nadiifinta shaybaarrada polar ama hydrophilic.Boostada dhexdeeda, peptide polar xoog leh ayaa loo adeegsaday muunad ahaan waxaana lagu sifeeyay tiirka C18AQ.Natiijo ahaan, badeecada bartilmaameedka ah ee buuxinaysa shuruudaha ayaa la helay waxaana loo isticmaali karaa cilmi-baarista soo socota iyo horumarinta.
| Qalabka | SepaBean™mashiinka 2 | |||
| Kartridges | 12 g SepaFlash C18 RP flash cartridge (silica wareegsan, 20 - 45 μm, 100 Å, Dalbo nunber：SW-5222-012-SP) | 12 g SepaFlash C18AQ RP flash card (silica wareegsan, 20 - 45 μm, 100 Å, Lambarka dalbashada: SW-5222-012-SP(AQ)) | ||
| Mawjada dhererka | 254 nm, 220 nm | 214 nm | ||
| Marxaladda mobilada | Walxaha A: Biyo Xalka B: Acetonitrile | |||
| Heerka socodka | 15 ml/min | 20 ml/min | ||
| Muunad rarista | 30 mg | |||
| Gradient | Waqtiga (CV) | Miraha B (%) | Waqtiga (min) | Miraha 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 | |||
Natiijooyinka iyo dood
Si loo barbar dhigo waxqabadka sifaynta ee muunada peptide polar inta u dhaxaysa tiirka C18 ee caadiga ah iyo tiirka C18AQ, waxaanu u isticmaalnay tiir C18 joogta ah nadiifinta toosh ee muunada bilow ahaan.Sida ku cad Jaantuska 2, sababta oo ah burburka wejiga hydrophobic ee silsiladaha C18 ee ay sababtay saamiga biyaha sare, muunada ayaa si dirqi ah loogu hayaa kartuunada caadiga ah ee C18 waxaana si toos ah uga baxay wajiga wareega.Natiijo ahaan, muunada si fiican looma kala saarin oo loo nadiifin.
Jaantuska 2. Karamatogaraamka toosh ee muunada ee ku jira kartoonka C18 ee caadiga ah.
Marka xigta, waxaan u isticmaalnay tiir C18AQ nadiifinta tooska ah ee muunadda.Sida ku cad Jaantuska 3, peptide si wax ku ool ah ayaa loogu hayaa tiirka ka dibna waa laga saaray.Badeecada la beegsaday ayaa laga soocay wasakhda ku jirta muunada ceeriin waana la ururiyay.Ka dib lyophilization ka dibna ay falanqeysay HPLC, badeecada la sifeeyay waxay leedahay nadiif ah 98.2% waxaana loo sii isticmaali karaa cilmi baarista iyo horumarinta talaabada xigta.
Jaantuska 3. Kormatogaraamka toosh ee muunada ee ku dul yaal kartoonka C18AQ.
Gebogebadii, SepaFlash C18AQ RP flash cartridge oo ay weheliso nidaamka chromatography-ga ee SepaBean™Mashiinku wuxuu bixin karaa xal degdeg ah oo wax ku ool ah oo loogu talagalay nadiifinta shaybaarrada polar ama hydrophilic.
Waxaa jira taxane ah kartoonada flash-ka ee SepaFlash C18AQ RP oo leh tilmaamo kala duwan oo ka yimid Tignoolajiyada Santai (sida ku cad Shaxda 2).
| Lambarka Shayga | Cabirka tiirka | Heerka socodka (ml/min) | Ugu badnaan.cadaadiska (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 |
Shaxda 2. SepaFlash C18AQ RP flash cartridges.Qalabka wax lagu baakadeeyo: Waxtarka spherical C18 (AQ) ee silica isku xiran, 20 - 45 μm, 100 Å.
Macluumaad dheeri ah oo ku saabsan tifaftirayaasha faahfaahsan ee mashiinka SepaBean™, ama macluumaadka dalbashada ee sepaFlash, fadlan booqo shabakadayada.
Waqtiga boostada: Oct-12-2018