4. Cell Separation - Clinical Research Products



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4. Cell Separation - Clinical Research Products Introduction of Dynabeads ClinExVivo 53 Dynabeads ClinExVivo Epoxy 54 Dynabeads ClinExVivo Sheep anti-mouse IgG 55 Visit for the latest product up-dates and references. 51

Application Overview Samples: Whole Blood Cord Blood Mononuclear Cells Bone Marrow Buffy Coat Spleen Lymph Node Tissue Digest Depletion Expansion Positive Isolation Negative Isolation DNA Isolation Organelle Isolation mrna Isolation Specific Protein Isolation Capture of Biotinylated Target 52

Introduction of Dynabeads ClinExVivo Cell Separation - Clinical Research Products Simple, rapid and reliable separation or stimulation of human cells as part of pre-clinical or phase I/II clinical trials in cell-based therapy. Introduction of Dynabeads ClinExVivo Applications Intended use Dynabeads ClinExVivo products are designed for use in pre-clinical or phase I/II clinical trials for cell-based therapy. They can be used during clinical research for ex vivo separation or stimulation of human cells. These versatile products can be easily coupled with your own antibodies or ligands. Cell separation is simple, rapid and reliable with high recovery efficiency. The viability and purity of the separated cells are excellent. 4 Quality Batch-to-batch variation is minimal when coupling antibodies or ligands to the monosized, polymer Dynabeads. This is due to a standardised surface area for coupling, which ensures consistent results in your clinical applications. Dynabeads ClinExVivo are produced under validated aseptic conditions. These strict conditions include: Aseptic processing in a fully validated Class 100 clean-room. Gamma-irradiated beads. Validated processes. Sterile buffers, vials and ligands (antibodies). Sterility testing according to United States Pharmacopeia. Endotoxin testing according to United States Pharmacopeia. Qualified operators. Certification Dynal Biotech is ISO 9001:2000 certified. US - A Master File is held with the United States Food & Drug Administration (FDA), which will assist users with applications for FDA approvals on clinical trials. If cross-referencing the Master File is of interest to an Investigational New Drug (IND) Application or other applications, please contact Dynal Biotech with the sponsor s and/or investigator s full name and address, along with project name and aim. Dynal Biotech will then be able to issue a Letter of Authorisation and this will enable the FDA to reference the Master File when reviewing the application. Fig.1: Aseptic processing in a fully validated Class 100 clean-room. Cell Separation - Clinical Research Products 53

Dynabeads ClinExVivo Epoxy 0434 Europe - The Dynabeads ClinExVivo products have been assessed by a Notified Body, Det Norske Veritas. Dynabeads ClinExVivo products conform to the requirements in the Medical Device Directive 93/42/EEC. The Certificates of Assessment EC are available upon request. Dynabeads ClinExVivo products are registered at the Norwegian Registry for Medical Devices, operated by the Norwegian Board of Health. This single registration is valid for all countries within the European Economic Area (EEA). The Council of European Union produces formal Directives covering a wide range of products intended for distribution within the EEA. The Medical Device Directive 93/42/EEC, in force from January 1995, has been mandatory since 13 June 1998. This directive describes the essential requirements as applied to Medical Devices and requires that any Devices intended for the European market must be CE marked. This marking, denoting Communautés Européennes, confirms that the Device complies with all relevant Directives. It must be affixed to Medical Devices that are marketed, distributed or sold anywhere in the EEA. The CE mark ensures free market admission without further controls. The Directive ensures free trade within the EEA, as well as safeguarding users and patients. The EC Declaration of Conformity places the ultimate responsibility for the Devices safety and product liability on the manufacturers. A Notified Body is an independent organisation appointed by a European national authority, to undertake conformity assessment before a product is CE marked. Det Norske Veritas (DNV) was appointed to be a Notified Body to carry out conformity assessment for Medical Devices 93/42/EEC on behalf of the European Union (EU). DNV approved Medical Devices should be marked as follows: 0434. Dynabeads ClinExVivo Epoxy (Product no. 402.01) Dynabeads ClinExVivo Epoxy are hydrophobic Dynabeads (diameter 4.5 µm) with activated epoxy groups on the surface. The epoxy group binds covalently to primary amino groups on proteins and peptides, and no further surface activation is required. The coupling reaction can take place at 4-37 o C for 16-20 hours at neutral ph, allowing the covalent binding of temperaturelabile antibodies. This product can be coated with the antibodies or ligands of your choice for pre-clinical or phase I/II clinical trials for cell based therapy. Applications Intended use Dynabeads ClinExVivo Epoxy can be used during pre-clinical work or phase I/II clinical trials. Coupled with ligands, such as antibodies, proteins, glycoproteins or carbohydrates, they can be used for the isolation, depletion or stimulation of specific human cells. 54

Dynabeads ClinExVivo Sheep anti-mouse IgG The required monoclonal antibodies (mab) or other ligands must be coupled to Dynabeads ClinExVivo Epoxy before use. For detailed instructions, please visit our website at. The coupling procedure and cell separation should be performed under aseptic conditions. Separation strategy: The cells can be separated using a direct technique such as depletion, negative or positive isolation (see page 11). Starting samples: Whole blood, bone marrow, umbilical cord blood, apheresis product, buffy coat and mononuclear cells (MNC). Certification For further information, please refer to the introduction of Dynabeads ClinExVivo products (page 53). EU - Dynabeads ClinExVivo Epoxy are CE 0434 marked and conform to the Medical Device Directive 93/42/EEC, class IIb. The Certificate of Assessment EC, issued by the Notified Body, Det Norske Veritas, is available upon request. US - A Master File is held with the FDA. The product contains: 10 ml of Dynabeads ClinExVivo Epoxy. Supplied at 4 x 10 8 beads/ml. Dynabeads ClinExVivo Sheep anti-mouse IgG (Product no. 422.01) Dynabeads ClinExVivo Sheep anti-mouse IgG are coated with polyclonal anti-mouse IgG and have a diameter of 4.5 µm. This product will bind the primary mouse IgG heavy and light chains, as well as the IgM light chain. No further surface activation is required. The coupling reaction takes place at 2-8 o C for 30 minutes with a gentle rotation. This product can be coated with the mouse IgG of your choice for pre-clinical or phase I/II clinical trials for cell based therapy. Applications Intended use Dynabeads ClinExVivo Sheep anti-mouse IgG can be used during pre-clinical work or phase I/II clinical trials. Using the required IgG mab, in combination with Dynabeads ClinExVivo Sheep anti-mouse IgG, ex vivo isolation, depletion or stimulation of the specific cell population can be conducted. For detailed instructions please visit our website at:. The coupling procedure and cell separation should be performed under aseptic conditions. Separation strategy: The cells can be separated by a direct or indirect technique, such as depletion, negative or positive isolation (see page 11). Starting samples: Whole blood, bone marrow, umbilical cord blood, apheresis product, buffy coat and MNC. Fig. 1: Scanning electron micrograph of an HIVinfected H9 cell isolated with anti-p24 coated Dynabeads. Courtesy of Prof. Dr. R.H. Dennin. 4 Cell Separation - Clinical Research Products 55

Dynabeads ClinExVivo Sheep anti-mouse IgG Certification For futher information, plese refer to the introduction to Dynabeads ClinExVivo products (page 53). EU - Dynabeads ClinExVivo Sheep anti-mouse IgG are CE 0434 marked and conform to the Medical Device Directive 93/42/EEC, class III. The Certificate of Assessment EC, issued by the Notified Body, Det Norske Veritas, is available upon request. US - A Master File is held with the FDA. The product contains: 10 ml of Dynabeads ClinExVivo Sheep anti-mouse IgG. Supplied at 4 x 10 8 beads/ml. Fig.1: Human hepatocyte isolated with Dynabeads. Courtesy of Dr. H. Danielsen. Dynabeads CustomDesign Products in Cellular Applications Enhance your ligand s performance Dynal Biotech has the competence and facilities to tailor-make a product for you. With Dynabeads CustomDesign products, our proprietary technology and know-how are made to work to your specifications. By customising the coupling of your ligand to Dynabeads, its performance in your application can be improved. For further details, see chapter 7. Dynal ClinExVivo Workstation Coming Soon! The Dynal ClinExVivo Workstation is intended for cellular research applications. It is a versatile, programmable device for medium to largescale magnetic cell separation based on the Dynabeads technology. The workstation combines the confidence of commercially-available closed disposable transfer bags and an integral programmable unit that allows you to control the temperature of your sample and vary the rotation speed during both the incubation and the magnetic separation steps. For further details, see chapter 8. Ordering Information Product Volume Product No. Dynabeads ClinExVivo Epoxy 10 ml 402.01 Dynabeads ClinExVivo Sheep anti-mouse IgG 10 ml 422.01 56