YMC Phases Preparative Scale. YMC Europe GmbH Friederike Becker
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1 YMC Phases Preparative Scale YMC Europe GmbH Friederike Becker
2 Production Facility Komatsu / Ishikawa Province Japan Courtesy of Courtesy YMC Europe of YMC GmbH Europe GmbH
3 YMC will never knowingly change any product that is under use in a validated production procedure or validated analytical method. Courtesy of YMC Europe GmbH
4 Analysis and Purification Natural Products Pharmaceuticals Bio- Pharmace uticals Peptides / Oligonucleotides Courtesy of Courtesy YMC Europe of YMC GmbH Europe GmbH
5 Analysis and Purification Silica Hybrid Polymer IEX Courtesy of Courtesy YMC Europe of YMC GmbH Europe GmbH
6 Hybrid phase YMC Innovations for (U)HPLC to Prep: YMC-Triart Ion exchange for proteins YMC-BioPro Silica based innovation: YMC-HG! Courtesy of YMC Europe GmbH
7 > 20 Fully Scalable Phases 15 mm N=4.600 P=250 psi 10 mm N=7.900 P=600 psi 5 mm N= P=2.600 psi Courtesy of YMC Europe GmbH
8 Selectivities in HG-Bulk RP: C18 / C8 / C4 / AQ Specialities: Phenyl / TMS NP: SIL / NH / Diol / CN Courtesy of YMC Europe GmbH
9 YMC*HG Bulk * Perfect spherical shape and smooth surface *chosen area is worst case and not representative for material Courtesy of YMC Europe GmbH
10 YMC*HG Bulk * No balloon particles *chosen area is worst case and not representative for material Courtesy of YMC Europe GmbH
11 Dynamic Mechanical Stability Courtesy of YMC Europe GmbH
12 YMC*HG vs. Conventional Silica Narrow particle size distribution Uniformity coefficient D 10 / D µm, 12 nm 15 µm, 20 nm conventional new Conventional new % better 30% better Courtesy of YMC Europe GmbH
13 YMC*HG Advantages Fast column packing Particle quality High overload capacity Pore size distribution Long lifetime = High productivity Courtesy of YMC Europe GmbH
14 YMC-Triart Versatile hybrid silica columns
15 Why multilayer hybrid particle? Selectivity ph & Temperature Robustness Courtesy of YMC Europe GmbH / YMC Co. Ltd.
16 Multilayer hybrid particle Inorganic siloxane phase Organic siloxane phase Multi-layered Hybrid Structure Courtesy of YMC Europe GmbH / YMC Co. Ltd.
17 Particle size (µm) Selectivities Triart C18 Triart C8 Diol-Hilic 1,9; 3; 5; 10; 20 um ph
18 Good Choice for MD Courtesy of YMC Europe GmbH / YMC Co. Ltd.
19 Stability at high ph Courtesy of YMC Europe GmbH / YMC Co. Ltd.
20 Flexibility in MD 1. Saccharin pka= Dextromethorphan pka:= Amitriptylen pka= Butylparaben pka= Ibuprofen pka=4.4
21
22 Separation of peptides at low ph
23 Structurally similar peptides at ph 7 ph 2.2 ph Column : YMC-Pack PolymerC18, 250 x 4.6 mmid Mobile phase : acetonitrile / water / TFA (18/82/0.05) (ph 2.2) Flow rate : 0.5 ml/min Detection : UV at 220nm Injection volume : 10µL Sample : 1. Angiotensin III 2. Angiotensin II 3. Angiotensin I Column : Triart C8-L (20 nm, 10 µm), 250 x 4.6 mmid Mobile phase A : acetonitrile / 10mM NH 4 H 2 PO 4 + NH 4 OH 50/50 (ph 7) Mobile phase B : 10mM NH 4 H 2 PO 4 + NH 4 OH (ph 7) Gradient : 35-65% A in 25min Flow rate : 1ml/min Detection : UV at 215nm Injection volume : 10µL Sample : 1. Angiotensin II 2. Angiotensin III 3. Angiotensin I
24 Mechanical Stability 1. Column packing Column size : 100 X 50 mm i. d. Packing pressure : 930 psi 2. Pressure measurement Eluent : methanol/water (85/15) Flow rate : 50 ml/min 3. Unpacking
25 Mechanical Stability Psi/m Hybrid silica for YMC-Triart Prep (10 mm, 200 Å) Conventional silica (10 mm, 200 Å) Packing number
26 Mechanical Stability 1. Packing 11. Packing Conventional silica (10 mm, 200 Å)
27 Mechanical Stability 1. Packing 11. Packing YMC-Triart (10 mm, 200 Å) Courtesy of YMC Europe GmbH / YMC Co. Ltd.
28 AU AU AU Chemical Stability 1st injection 20th injection After regeneration M NaOH min min min
29 AU AU AU AU AU AU AU Initial After 10 cleaning cycles After 35 cleaning cycles After 70 cleaning cycles YMC-Triart Prep C8-S (10 mm, 20 nm) min min min Good peak shape min Initial After 10 cleaning cycles After 35 cleaning cycles Conventional silica based C8 (10 mm, 20 nm) min min Peak broadening! min Extended lifetime!
30 Number of possible peptide production cycles Competitor A-1 Competitor A-2 YMCbasic Competitor B YMC-Triart Prep C8-L Confidential Courtesy of YMC Europe GmbH
31 YMC-Triart Prep C18/C8-S Scalable material (1,9 µm, 3 µm, 5 µm, 10 µm, 15 µm 20 µm) 100% aqueous mobile phase possible Cleaning with 0.1 M NaOH possible
32 YMC-Triart QA
33 YMC-Triart Multilayered hybrid particles ph & temperature stability Versatile selectivity UHPLC
34 YMC-BioPro Designed for IEX Proteins and Peptides Preparative Scale Economic Gel
35 YMC-BioPro preparative scale YMC-BioPro Q YMC-BioPro S Bead hydrophilic polymer beads Particle size 5 µm, 10 µm, 30 µm, 75 µm Charged group -CH 2 N + (CH 3 ) 3 -(CH 2 ) 4 SO - 3 ph range
36 YMC-BioPro preparative scale Features Excellent dynamic binding capacity (DBC) Low non-specific adsorption and high recovery High capacity with low operating pressure even at high flow rates Scalable from analytical to prep
37 DBC at different flow rates High DBC even at high flow rates for rapid large-scale purification!
38 CIP study of BioPro S75 Separation of proteins Determination of DBC and Recovery CIP with 1.0 M NaOH (5CV at 400 cm/hr)
39 DBC (mg/ml-resin) Recovery () CIP study of BioPro S DBC Numbers of CIP with 1.0 M NaOH
40 CIP study of BioPro S75
41 DBC (mg/ml-resin) YMC-BioPro QA BSA Trypsin inhibitor IgG M.W. (X 10 5 ) Protein MW kda DBC Eluted amount (mg/mlresin) (mg/mlresin) Recovery (%) IgG BSA Try. I
42 DBC (mg/ml-resin) YMC-BioPro SP IgG Avidin Lysozyme M.W. (x 10 5 ) Protein MW kda DBC Eluted amount (mg/mlresin) (mg/mlresin) Recov ery (%) IgG Avidin Lys
43 Cation-exchange purification of N,N-bis-methionyl-histone Н1.3 YMC-BioPro S-75, 120 x 25 mmid, V = 60 ml DBC Dr. Gusarov et al., Russian Journal of Biopharmaceuticals, Vol 2, No 1 (2010)
44 Cation-exchange purification of N,N-bis-methionyl-histone Н1.3 Sorbent SP-sepharose FF BioPro S-75 Macro-prep HS-100 Matrix Polysaccharide Polymethacrylate Polystytrene Particle size, um Porosity, nm DBC by histone, g/l Yield, % Purity (HPLC), % Dr. Gusarov et al., Russian Journal of Biopharmaceuticals, Vol 2, No 1 (2010)
45 Human Serum Protein YMC-BioPro QA 5mm, 50 x 4.6 mm ID High resolution of transferrin forms. TSKgel BioAssist Q 10 mm, 50 x 4.6 mm ID 10mm, 50 x 5.0 mm ID GE MonoQ 10 mm, 50 x 4.6 mm ID Eluent: A) 20 mm Tris-HCl; B) 20 mm Tris-HCl containing 0.5 M NaCl; Flow rate: 0.5 ml/min; Temperature: 25 C; Detection: UV at 280 nm; Injection: 20 ml; Sample: Human serum (100 ml/ml)
46 mau Client data: YMC-BioPro for mab-qc (analytical) DAD1 E, Sig=280,10 Ref=360,100 (MR24MAR09\ \MR24MAR D) DAD1 E, Sig=280,10 Ref=360,100 (MR24MAR09\ \MR24MAR D) 80 YMC BioPro SP-F 60 Dionex ProPac WCX min
47 Client data: YMC-BioPro for mab-qc (analytical) Serial # s
48 Quality Management DIN EN ISO 9001:2008 (Production & Sales) Drug Master File for standard phases available (YMC*Gel, YMC-BioPro) Regulatory Support File for YMC-BioPro GMP/FDA compliance available for Pilot columns
49 Drug Master File / Regulatory Support File
50 Efficient Preparative Applications of YMC-BioPro Proteins & Antibodies Chymotrypsinogen A Herceptin Filgrastim Histones Interferon Factor VIII IgY, IgG Variants of mabs
51 Selection of References for YMC*Gel & YMC-BioPro Eli Lilly Novo Nordisk Novartis Sandoz PolyPeptide Laboratories Lek Bachem Bayer Schering Pharma Ipsen Boehringer Ingelheim Sigma Aldrich TEVA/API Cilag American Peptides Genzyme Minapharm Carbogen Roche AstraZeneca Gedeon Richter Bayer Hyglos Rentschler Chromacon Masterclon IBCH Moscow Roche
52 Preparative columns Inner diameter: mm Length: mm Semi-prep columns Inner diameter: 50 mm Length: mm Courtesy of YMC Europe GmbH
53 Courtesy of YMC Europe GmbH
54 Glass Pilot columns Courtesy of YMC Europe GmbH
55 Facilities in Japan Courtesy of YMC Europe GmbH
56 Customised stationary phases TQP12S21 minus TOS08S11 TAS08S11 Higher loadability and resolution OMG99S11 Better resolution
57 YMC is certified according to DIN ISO 9001 Drug Master File for standard phases available Courtesy of YMC Europe GmbH
Stability Axial Compression Technology Application Ordering Information
42 YMC-Actus YMC-Actus 43 YMC-Actus Contents Stability... 44 Axial Compression Technology... 45 Application...46-47 Ordering Information...48-49 Introduction Fast semi-preparative chromatography Semi-preparative
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