Polyethylene glycol(PEG)因其生物相容之特性，廣泛應用於藥物、生物醫藥及產業領域。常見於胜肽、蛋白質及其他治療藥物之應用。對於PEGylated protein之特性分析可能非常複雜並且需要使用各種不同的方法。
YMC於美國與Pelican Expression Technology實驗室合作開發出使用YMC-BioPro陽離子交換樹脂管柱進行PEGylated protein charge-variant之應用，展現出YMC BioPro管柱為此應用提供一穩定且再線性佳之分析工具，並優於目前分析charge-variant之方法。
更多詳細內容請參考LCGC雜誌內容：
The Utility of Cation Exchange Column for Successful Charge-Variant Analysis of PEGylated Proteins
填充管柱後，建議在進行樣品純化之前，使用標準樣品進行理論塔板數（N, number of theoretical plates）、理論塔板高度（HETP, height equivalent to a theoretical plate）和峰不對稱因子（As, peak asymmetry factor）測試及計算。藉由定期和重複執行此類標準測試，可以有效地評估膠體(gel, packing material)的品質和耐用性。 如果您想比較各個測試之間的結果，建議在每次測試中在相同條件下使用相同的測量設備。測量結果可能會受到一些因素影響，例如：死體積（dead volume）、流速（flow rate）、溫度（temperature）、沖提液（eluent）和進樣量（sample loading amount）的差異。
下列公式可用於計算理論塔板數（N, number of theoretical plates）。 較大的 N 值對應於層析管柱中較高的填充膠體密度。 此外，在固定滯留時間（retention time），峰形越窄，N 值越高；柱長越長，粒徑越小，N 值越大。 通常，每柱床長（m）的理論塔板數通常用作評估層析管柱純化性能的指標。
N = 5.54 x (tR / W0.5h)2
N/m ＝ N / L
L ＝ Column bed length (m)
理論塔板高度 （HETP, height equivalent to a theoretical plate）有時也用作評估層析管柱性能的指標。 下列公式可用於計算 HETP。 該值越小，填充在柱中的膠體密度越高。
HETP ＝ L / N
洗脫峰形 （peak shape）也是評估層析管柱性能的重要因素。 峰不對稱因子 (As) 的測定是一種相對簡單的估計層析管柱性能的方法。 下列公式可用於計算 As，該值越接近“1”是越理想的，其表示越高的峰形對稱性。
As ＝ b / a
我應該平衡HPLC管柱多久時間呢？
在執行梯度條件後進行下一次進樣前之HPLC層析管柱平衡時，如果（重新）平衡時間太短，則會出現實驗重複性不佳的風險。如果時間太長，則會浪費時間和溶劑。
在本HPLC技術指引中，您將學習如何通過計算管柱體積來估算最佳平衡體積。此外，還使用了一個實際案例來顯示不同平衡體積對分析結果的影響。有關管柱平衡的詳細說明，您可以在此處下載HPLC技術指引。
您的(U)HPLC層析管柱之分離效果是否開始變差？
這份文件我們提供了合適的管柱清洗程序，有助於恢復層析管柱之性能。此外，該程序還可防止污染物在固定相上累積。但是，需考量一些情況，例如：清潔溶劑的類型和有效再生所需的體積。有關層析管柱再生的更多詳細信息，請在此處下載說明文件。
Do you analyse hydrophilic compounds? Do you want to improve the retention on polar stationary phases such as PFP or Diol?
In this technical note, two different options for the improved analysis of hydrophilic compounds on YMC-Triart PFP and YMC-Triart Diol-HILIC are shown. The pH value and the amount of organic solvent are examined to learn more about the retention behaviour of basic, acidic and zwitterionic compounds using these phases. Due to their different selectivity, pentafluorophenyl (PFP) or Diol modifications are an ideal choice for the analysis of hydrophilic compounds which often remain unretained or unresolved on conventional C18 phases.
You can download the full method details here.
To get further information about the YMC-Triart series of columns.
As the COVID-19 pandemic continues, the worldwide search for an effective treatment is still ongoing. Remdesivir and its active metabolite GS-441524 are both antiviral drugs that have been examined and partially approved for the treatment of COVID-19 due to their capability of inhibiting virus replication.
In this application note, the separation of GS-441524 and Remdesivir in only 3 minutes is shown. A YMC-Triart C18 UHPLC column provides fast and reliable results making it an ideal choice for QC purposes.
You can download the application note with the full method details here.
The full article can be downloaded as a pdf.
| USP Class | Functional group | USP Description | YMC Product |
| L01 | C18 | Octadecyl silane <ODS or C18> chemically bonded to porous silica or ceramic particles – 1.5 to 10 μm in diameter |
|
| L03 | SIL | Porous silica particles – 1.5 to 10 μm in diameter | |
| L07 | C8 | Octyl silane <C8> chemically bonded to porous silica particles – 1.5 to 10 μm in diameter | |
| L08 | NH2 | An essentially monomolecular layer of aminopropylsilane <NH2> chemically bonded to totally porous silica gel support – 1.5 to 10 μm in diameter | |
| L10 | CN | Nitrile groups <CN> chemically bonded to porous silica particles – 1.5 to 10 μm in diameter | |
| L11 | Ph | Phenyl groups chemically bonded to porous silica particles – 1.5 to 10 μm in diameter | |
| L13 | C1 | Trimethylsilane <C1> chemically bonded to porous silica particles – 1.5 to 10 μm in diameter | |
| L20 | Diol | Dihydroxypropane groups chemically bonded to porous silica particles – 1.5 to 10 μm in diameter | |
| L26 | C4 | Butyl silane <C4> chemically bonded to porous silica particles – 1.5 to 10 μm in diameter | |
| L27 | SIL | Porous silica particles – 30 to 50 μm in diameter | |
| L33 | Diol | Packing having the capacity to separate dextrans by molecular size over a range of 4,000 to 500,000 Da. It is spherical, silica-based, and processed to provide pH stability | |
| L40 | Cellulose tris-3,5- dimethylphenylcarbamate |
Cellulose tris-3,5-dimethylphenylcarbamate coated porous silica particles, 3 to 20 µm in diameter | |
| L43 | PFP | Pentafluorophenyl groups chemically bonded to silica particles by a propyl spacer, 1.5 to 10 µm in diameter | |
| L51 | Amylose tris-3,5- dimethylphenylcarbamate |
Amylose tris-3,5-dimethylphenylcarbamate-coated, porous, spherical, silica particles, 3 to 20 µm in diameter | |
| L59 | Diol | Packing for the size exclusion separations of proteins (separation by molecular weight) over the range of 5 to 7000 kDa. It is spherical (1.5 to 10 μm), silica or hybrid packing with a hydrophilic coating | |
| L62 | C30 | C30 silane bonded phase on a fully porous spherical silica, 3 to 15 µm in diameter | |
| L99 | Amylose tris-3,5- dimethylphenylcarbamate |
Amylose tris-(3,5-dimethylphenylcarbamate), immobilized on porous, spherical, silica particles, 3 to 5 µm in diameter | |
| L111 | Polyamine | Polyamine chemically bonded to porous spherical silica particles, 5 μm in diameter | |
| L119 | Cellulose tris-3,5- dichlorophenylcarbamate |
Cellulose tris-(3,5-dichlorophenylcarbamate), immobilized on porous, spherical, silica particles, 3 to 5 µm in diameter. |
YMC玻璃管柱可針對客戶實際應用進行最佳化選擇，對於藥物與蛋白質純化日益複雜且高純度的需求，除有多種內徑、可調式長度選擇之外，亦可針對您所需之耐壓、溫控、耐溶劑等實驗需求提供相對應的產品。並且可供應從實驗室一路放大到生產端之完整的產品線供應。以下為簡單選擇指引，詳細規格請參考各產品連結或來電與我們諮詢。
-低~中壓應用
-管柱內徑: 10, 15, 25, 50, 70, 80 mm
-管柱長度: 120, 200, 450, 750, 1000 mm(管柱長度可自由調整)
-耐壓: 5 ~ 30 bar(依內徑大小耐壓規格略有差異)
–溫控夾套(選配): Heating/cooling jacket
-操作快速: Easy screw-lock 設計操作便利
-中~高壓應用
-管柱內徑: 5, 10, 15, 25, 35, 50 mm
-管柱長度: 125, 250, 500 mm(管柱長度可自由調整)
-耐壓: 15 – 80 bar(依內徑大小耐壓規格略有差異)
-再現性佳
-操作快速: Quick-Lock 設計操作便利
-中壓應用
-管柱內徑: 10, 15, 25, 50 mm
-管柱長度: 120, 200, 300, 450, 750, 1000 mm(管柱長度可自由調整)
-操作溫度: 4 – 70 °C
-耐壓: 30 – 10 bar(依內徑大小耐壓規格略有差異)
-操作快速: Easy screw-lock 設計操作便利
-應用廣泛: 適用於BioLC或逆相(RP)/正相(NP)等應用
-內徑: 100, 140, 200, 300 mm
-可訂製不繡鋼材質
-可選配溫控夾套(Heating/Cooling jacket)
-操作簡單、密封(Seal)技術佳
(更大內徑需求請來信詢問)
-Bio液相層析之生物相容性
-內徑: 70, 100, 140, 200 mm
-特殊排液設計使填充管柱簡單且無氣泡產生
-可拆卸式玻璃管柱設計可快速且乾淨的卸除玻璃管柱
| 小分子化合物
Low molecular weight compounds |
一般有機化合物
General organic compounds |
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| General organic compounds | YMC-Triart C18 Meteoric Core C18 |
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| Chemical structures differ slightly among solutes | YMC-Triart C18 ExRS | ||
| Hydrophobicity differs greatly among solutes | YMC-Triart Bio C18 J’sphere ODS-L80 |
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| Difficult to separate using standard ODS | YMC-Triart C18 ExRS Hydrosphere C18 YMC-Pack Pro C18 RS YMC-Pack ODS-AL J’sphere ODS-H80 J’sphere ODS-L80 YMC-Pack PolymerC18 |
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| Basic compounds | YMC-Triart C18 YMC-Triart C18 ExRS |
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| Coordination compounds | YMC-Triart C18 YMC-Triart C18 ExRS |
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| Sugars and glycosides | YMC-Triart C18 Hydrosphere C18 |
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| High molecular weight compounds
大分子化合物 |
High molecular weight peptides and oligonucleic acids | YMC-Triart C18 YMC-Triart Bio C18 Meteoric Core C18 BIO |
|
| Alkaline mobile phase
鹼性移動相條件 |
YMC-Triart C18 YMC-Triart C18 ExRS YMC-Triart Bio C18 YMC-Pack PolymerC18 |
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