Cellufine MAX HIC Butyl, Phenyl Technical Data Sheet

Transcription

1 Hydrophobic Interaction Chromatography Media MAX HIC Butyl, Technical Data Sheet 2-1, temachi 2-Chome Chiyoda-ku, Tokyo JAPAN Phone / Fax cellufine@jnc-corp.co.jp web: http: //

2 Technical DATA Sheet MAX HIC ( and Butyl) High Flow Rate typed Media MAX is a 2nd generation media with high flow characteristics. JNC developed a new, highly cross-linked base resin for MAX series. MAX hydrophobic interaction chromatography is now available with and Butyl chemistries. MAX base resin Cellulose, a natural polysaccharide, possesses unique crystalline molecular structure differing from noncrystalline polysaccharides such as agarose. Thus has distinctive pore structure as shown in the pictograph (Fig. 1). The new MAX series offers the largest pore size of all chromatography media. The benefit of such pore size in MAX HIC media provides superior strength and excellent mass transfer. Characteristics of MAX HIC Media The basic characteristics of MAX HIC media are shown in Table 1. All MAX HIC media are based on 9 μm (average) highly cross-linked cellulose beads. MAX HIC media are designed for use in bio-pharmaceuticals manufacturing processes. Matrix Particle size Highly Cross-linked Cellulose ~13 μm Ligand type Butyl BSA adsorption capacity (mg/ml) BSA elution efficiency (%) Polyclonal IgG 1% DBC (mg/ml) perating pressure MAX LS <.3 MPa ph stability ph 2 ~ 13 Storage % ethanol Table 1. Characteristics of MAX HIC media Ligand content is controllable in MAX HIC media. JNC is able to optimize both and media on request as indicated in Figure BSA adsorption ( ) and recovery ( ) 1 Fig 1. SEM analysis of MAX base resin Partial structure of MAX HIC Ligand structure for MAX HIC media are described in Fig.2. BSA adsorption(mg/ml) Recovery (%) MAX LS R H Ligand contents 1 LS MAX Butyl H Fig.2. Ligand Structure of MAX HIC R Fig.2. Relationships between ligand contents and BSA adsorption capacity or protein recovery in. 2 IN-MAX HIC-MC-EN Ver.3.1 (1-July-1)

3 Pressure-flow Properties of MAX HIC Media MAX HIC media enable high-flow operation, which is essential to efficient purification of biopharmaceuticals. The figures below show pressure-flow velocity curves of MAX HIC media (Fig. 3). All MAX HIC media are operable at practical flow velocities and pressures. Column: 6.6 mm ID 5 mm L Buffer A: 1 mm phosphate buffer (ph 7) M (NH ) 2 S Buffer B: 1 mm phosphate (ph 7) Proteins: Ribonuclease A, Cytochrome C, Lysozyme Model Protein Adsorption for MAX Pressure (MPa) MAX MAX phenyl6 Mab select 1 3 Flow velocity (cm/h) Adsorption capacity (mg/ml) poly IgG Column: 5 mm ID x 1 cm L BSA concentration: 1 mg/ml : LS : M NaCl 1M NaCl BSA Buffer: 5 mm Tris-HCl (ph 8.5) + NaCl Fig 3. Pressure-flow velocity curves of and (2.2 cm I.D. x cm L), Mobile phase: pure water at 2. Model Protein Separation Performance for HIC media MAX HIC media are optimized for high resolution. Fig. shows Model protein separation with (standard and LS) and. Protein separation studies show that relative binding strength are > LS >. BSA 1% DBC(mg/ml) Fig.5. Salt Concentration vs. Model Protein Adsorption of Dynamic Binding Capacity for MAX vs. Salt Concentration LS LS 1 M (NH ) 2 S 1.5 M (NH ) 2 S 2 M (NH ) 2 S Column: 5 mm ID x 5 mm L Flow velocity: 15 cm/hr BSA concentration: 1 mg/ml Buffer: mm Phosphate (ph7) + (NH ) 2 S Fig.6. Salt Concentration vs. BSA-DBC of Fig. Model Proteins Separation for and 3 IN-MAX HIC-MC-EN Ver.3.1 (1-July-1)

4 Polyclonal IgG ) poly IgG 1% DBC (mg/ml) LS Chemical Stability and Cleaning-In-Place Cellulose is well-known as a natural product having chemical and physical stability. Thus, since is derived from cellulose, it also is stable to chemicals and caustic and acidic solutions. CIP of all MAX HIC media can be carried out with.5 M NaH solution. Used media should be stored in % ethanol at 2-25 after cleaning..5 M (NH ) 2 S Column: 5 mm ID x 5 mm L Flow velocity: 15 cm/hr 1 M (NH ) 2 S IgG concentration: 1 mg/ml Buffer: mm Phosphate (ph 7) + (NH ) 2 S Fig.7.. Polyclonal IgG-DBC of at different salt concentrations Ethanol (7%) Isopropyl alcohol (3%) Guanidine hydrochloride (6M) Urea (6M) NaH (.5M) Detergents Autoclave (121 ºC, min) Dynamic Binding Capacity for MAX Butyl 1 % DBC (mg/ml) 3 1 BSA IgG Column: 5 mm ID x 5 cm L Flow rate:.5 ml/ min Buffer: 1 mm Phosphate (ph 7.) + 2 M (NH ) 2 S / BSA 1 M (NH ) 2 S / polyclonal IgG Fig.8. DBC of Efficient mass-transfer characteristics of MAX HIC media translate to superior dynamic binding capacities (DBC). Figures 6 to 8 show DBC of model proteins for MAX HIC media. MAX HIC media are suitable for use in down-stream steps in bio-pharmaceutical purification. IN-MAX HIC-MC-EN Ver.3.1 (1-July-1)

5 rdering Information Product Name MAX Pack Size 1 ml 5 ml 1 lt 1 ml 5 ml 1 lt Catalogue No Product Name LS Pack Size 1 ml 5 ml 1 lt Catalogue No Technical Support contacts (North & South America, Europe) JNC America Incorporated 555 Theodore Fremd Avenue, Suite C-6 Rye, NY 158 USA TEL: FAX: cellufine@jncamericany.com Please Send Purchase rders to: (North America & Europe) JNC America Incorporated 555 Theodore Fremd Avenue, Suite C-6 Rye, NY 158 USA TEL: FAX: cellufine@jncamericany.com (Asia & thers) JNC Corporation 2-1, temachi 2-Chome, Chiyoda-ku Tokyo Japan Tel: Fax: cellufine@jnc-corp.co.jp (Asia & thers) JNC Corporation 2-1, temachi 2-Chome, Chiyoda-ku Tokyo Japan Tel: Fax: cellufine@jnc-corp.co.jp Please visit our web site for more information http: // 5 IN-MAX HIC-MC-EN Ver.3.1 (1-July-1)