A New Generation of Matrix for Protein Purification Offers 3 High (3H)
A New Standard and Criteria for Chromatography Media
μSphere™微球是與Source相同的層析填料基材，是集高流速、高載量、高分辨率(3H)于一體的新一代蛋白純化填料的基礎材料。過去二十多年來，聚合物微球得到了極大發展并成功應用于生物物質分離和純化。PAE開發的聚合物微球，簡寫μSphere™是用單分散高交聯度聚丙烯酸酯微球為基材并在其表面加特殊涂層并接枝功能基團或配體制備的層析介質。傳統的多糖基基材雖生物相容性好，但其骨架剛性低，在較高的壓力下容易出現變形或者孔道的改變，影響純化效率。μSphere™有效解決了瓊脂糖骨架剛性低的問題，具有諸多更適于線性放大，加快工藝開發速度。不但適于實驗室研究用樣品的制備，而且是工業規模純化蛋白、多肽、核酸、抗生素和低分子量藥物不可回避的選擇，且 PAE生產的用于生物大分子分離的微球為多孔的，有多種粒徑和孔徑，可達1000A,根須需求接上-OH,-COOH或-NH2 等基團，一邊方便偶聯其它配體或配基。粒徑/孔徑默認50mm，粒徑800 ?。可按客戶要求提供粒徑15mm，30mm，100mm和孔徑300、500和800的產品。
高流速
μSphere具有很高的機械穩定性且粒徑單一，因此層析特性較好，具有極很好的抗壓縮性能，不易出現堵塞。另外，其可耐操作過程中高至10MPa的壓力，支持高達2000cm/hour的流速。
高載量
μSphere是一種可精確控制孔徑和粒徑的單分布多孔微球體，其比表面積高達>500m2/g,用其制備的層析填料具有很高的功能基團密度，其動態載量。圖1是NTA-μSphere和NTA-Sepharose FF對比15個His-標簽蛋白的載量分布。圖中藍色柱代表NTA-μSphere對某樣品的載量占NTA-Sepharose FF載量的倍數，紅線是NTA-Sepharose FF的相對載量等高線，可見NTA-μSphere顯著超過NTA-Sepharose FF。
高分辨率
μSphere的粒徑相對較小(50μm±5%)且分布相當均一，機械強度高，不易變形，因此分辨率明顯高于糖基微球。圖2是NTA-μSphere和NTA- Sepharose FF上樣量等條件均一條件下的洗脫峰，顯然，NTA-μSphere的峰形更"sharp"。
其他特點
除上述3H外，mSphere在pH1到14都是穩定的，因此可以用0.5 MNaOH甚至更高的堿濃度進行在位清洗(CIP)。堿液在位清洗是科研產品除熱源以及雜蛋白的方法。
Appendix
下表是mSphere與Agarose微球與蛋白純化有關的理化性質的比較。從中可以看出，mSphere具有多方面的優勢。
主要基材類型總結與對比
類型 | 基材 | 粒徑(μm） | pH穩定性 | 壓力 | 流速(cm/h) |
Source 15 | PS/DVB | 15 | 1~14 | 10MPa | 900 |
Source 30 | PS/DVB | 30 | 1~14 | 10 | 2000 |
Resource | PS/DVB | 15 | 1~14 | 10 | 1800 |
MonoBeads | Monodisperse porous PS/DVB | 10 | 1~14 | 10 | 1200 |
MiniBeads | Monodisperse PS/DVB | 3, non porous | 1~14 | 10 |
|
Fast Flow | 6% cross-linked agarose | 45-165, avr. 90 | 2~13 | 0.3 | 300 |
Capto | Highly cross-linked agarose with dextran surface extender | 90 (d50V) | 2~13 | 0.3 | 1200 |
HP | 6% cross-linked agarose | 24-44, avr. 34 | 1~14 | 0.3 | 150 |
mSphere | Polyacrylate or PS/DVB | 3, 10, 15, 30, 50, 100 | 1~14 | 10 | 1200~2000 |
IMAC-μSphere
This is an affinity purification matrix for His-Tag Fusion proteins and other metal proteins.
polymer μSphere-based chromatographic matrix High Flow Rate，High capacity and high performance
IMAC μSphere 6 cess IMAC medium, designed for purifying proteins and peptides that have an affinity for metal ions, such as histidine-tagged proteins but also untagged recombinant or native proteins
l Uncharged IMAC resin allowing possibility to charge with your metal ion of choice for optimized selectivity
l Convenient purification of histidine-tagged proteins, also when Ni2+ is not the best choice of metal ion
l As BioProcess medium it fulfills industrial demands for security of supply, robust performance and regulatory support.
l Available in the convenient and time-saving prepacked HiTrap and HiPrep formats.
IMAC μSphere 6 is a member of the range of IMAC chromatography media for capture and intermediate purification.
IMAC μSphere 6 is composed of highly cross-linked beads modified with a novel chelating ligａnd immobilized to the base matrix. IMAC μSphere 6 is supplied free of metal ions, allowing the user to charge it with the most appropriate metal ion for purification of a target protein.
The coupling chemistry and the novel ligａnd is designed to give minimal ligａnd leakage while providing high capacity.
IMAC μSphere 6 is available in a range of different bulk pack sizes and convenient pre-packed formats for easy scale-up and process development.
As member of the BioProcess media range, IMAC μSphere 6 meets industrial demands with security of supply and comprehensive technical and regulatory support
Appendix:
Summary and Comparison of Matrices Extensively and Successfully Used in Protein Purification and Analysis
Category | Matrix | Particle size (μm） | pH Stability | Max Pressure | Max Flow Rate(cm/h) |
Source 15 | PS/DVB | 15 | 1~14 | 10MPa | 900 |
Source 30 | PS/DVB | 30 | 1~14 | 10 | 2000 |
Resource | PS/DVB | 15 | 1~14 | 10 | 1800 |
MonoBeads | Monodisperse porous PS/DVB | 10 | 1~14 | 10 | 1200 |
MiniBeads | Monodisperse PS/DVB | 3, non porous | 1~14 | 10 |
|
Fast Flow | 6% cross-linked agarose | 45-165, avr. 90 | 2~13 | 0.3 | 300 |
Capto | Highly cross-linked agarose with dextran surface extender | 90 (d50V) | 2~13 | 0.3 | 1200 |
HP | 6% cross-linked agarose | 24-44, avr. 34 | 1~14 | 0.3 | 150 |
mSphere | Polyacrylate or PS/DVB | 3, 10, 15, 30, 50, 100 | 1~14 | 10 | 1200~2000 |
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