CE-IVD reagents

Support for GxP processes in cell manufacturing

Germany/Austria/ SwitzerlandMiltenyi Biotec GmbH Friedrich-Ebert-Straße 68 51429 Bergisch Gladbach Germany Phone +49 2204 8306-0 Fax +49 2204 85197 macs@miltenyibiotec.de

USA/CanadaMiltenyi Biotec Inc. 2303 Lindbergh Street Auburn, CA 95602, USA Phone 800 FOR MACS Phone +1 530 888 8871 Fax +1 877 591 1060 macs@miltenyibiotec.com

AustraliaMiltenyi Biotec Australia Pty. Ltd. Unit 16 A, 2 Eden Park Drive Macquarie Park NSW 2113 Australia Phone +61 2 8877 7400 Fax +61 2 9889 5044 macs@miltenyibiotec.com.au

BeneluxMiltenyi Biotec B.V. Sandifortdreef 17 2333 ZZ Leiden The Netherlands macs@miltenyibiotec.nlCustomer service The NetherlandsPhone 0800 4020120 Fax 0800 4020100Customer service BelgiumPhone 0800 94016 Fax 0800 99626Customer service LuxembourgPhone 800 24971 Fax 800 24984

ChinaMiltenyi Biotec Technology & Trading (Shanghai) Co., Ltd. Rooms 2303 and 2309 No. 319, Xianxia Road Changning District 200051 Shanghai, P.R. China Phone +86 21 62351005 Fax +86 21 62350953 macs@miltenyibiotec.com.cn

FranceMiltenyi Biotec SAS 10 rue Mercoeur 75011 Paris, France Phone +33 1 56 98 16 16 Fax +33 1 56 98 16 17 macs@miltenyibiotec.fr

ItalyMiltenyi Biotec S.r.l. Via Paolo Nanni Costa, 3040133 Bologna Italy Phone +39 051 6 460 411 Fax +39 051 6 460 499 macs@miltenyibiotec.it

JapanMiltenyi Biotec K.K. Nittsu-Eitai Building 5F 16-10 Fuyuki, Koto-ku, Tokyo 135-0041, Japan Phone +81 3 5646 8910 Fax +81 3 5646 8911 macs@miltenyibiotec.jp

Nordics and BalticsMiltenyi Biotec Norden AB Scheelevägen 17 223 70 Lund Sweden macs@miltenyibiotec.seCustomer service SwedenPhone 0200-111 800 Fax 046-280 72 99 Customer service DenmarkPhone 80 20 30 10 Fax +46 46 280 72 99 Customer service Norway, Finland, Iceland, and Baltic countriesPhone +46 46 280 72 80 Fax +46 46 280 72 99

SingaporeMiltenyi Biotec Asia Pacific Pte Ltd. 100 Beach Road #28-06 to 28-08 Shaw Tower Singapore 189702 Phone +65 6238 8183 Fax +65 6238 0302 macs@miltenyibiotec.com.sg

South KoreaMiltenyi Biotec Korea Co., Ltd Arigi Bldg. 8F 562 Nonhyeon-ro Gangnam-gu Seoul 06136, South Korea Phone +82 2 555 1988 Fax +82 2 555 8890 macs@miltenyibiotec.co.kr

SpainMiltenyi Biotec S.L. C/Luis Buñuel 2 Ciudad de la Imagen 28223 Pozuelo de Alarcón (Madrid) Spain Phone +34 91 512 12 90 Fax +34 91 512 12 91 macs@miltenyibiotec.es

United KingdomMiltenyi Biotec Ltd. Almac House, Church Lane Bisley, Surrey GU24 9DR, UK Phone +44 1483 799 800 Fax +44 1483 799 811 macs@miltenyibiotec.co.uk

www.miltenyibiotec.com

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Miltenyi Biotec provides products and services worldwide. Visit www.miltenyibiotec.com/local to find your nearest Miltenyi Biotec contact. Front cover image ‘Eye’ copyright © Miles Away Photography / Shutterstock.com.

CE-IVD reagents are for in vitro diagnostic use, and are not intended for human in vivo applications. In the EU, CE-IVD reagents are available as CE marked in vitro diagnostic medical devices and meet the requirements of the European directive 98/79/EC on in vitro diagnostic medical devices (IVDD). CE-IVD reagents are designed and manufactured under the ISO 13485 certified quality system. These products are not available for use in the United States. MACS and the MACS logo are registered trademarks or trademarks of Miltenyi Biotec GmbH and/or its affiliates in various countries worldwide. Copyright © 2018 Miltenyi Biotec GmbH and/or its affiliates. All rights reserved.

Designed and manufactured according to quality management system ISO 13485

Fulfilling essential regulatory requirements for in vitro diagnostic medical devices

Optimized for reliable in-process and quality control in cell manufacturing and translational workflows

In order to enable our customers to meet regulatory guidelines, we provide CE-IVD reagents for clinical flow cytometry.

These reagents are in vitro diagnostic (IVD) medical devices that are in compliance with the European Directive 98/79/EC and designed and manufactured according to the quality management system ISO 13485. This involves highly controlled manufacturing processes, validated testing procedures, and an extensive documentation level. The resulting CE-IVD quality ensures:

• High level of documentation

• High level of in-process controls

• Conformity with European Directive 98/79/EC prior to first sale

• Quality audits by notified bodies after market introduction

What is a CE-IVD reagent?

CE-IVD reagents in translational workflows

Working in cell manufacturing? Quality control of cellular products for advanced therapies require a high level of standardization, which strongly depends on consistent reagent performance. CE-IVD reagents not only provide a high-quality standard for incoming goods inspection, in-process control, and release tests, but also offer the following advantages over research use only (RUO) reagents:

• Compliance with regulatory requirements when applying for clinical trials

• Lot-to-lot consistency for increased reproducibility over time

• Reduced in-house efforts during assay validation

*IPC/QC – incoming goods inspection, in-process control and release test

IN-PROCESS CONTROLCell manufacturing02

DONOR

INCOMING GOODS INSPECTIONBloodBone marrow Leukapheresis

01PATIENT

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In-process control and quality control (IPC/QC*) during cell manufacturing

CE-IVD reagents

The CE-IVD reagents we provide are ready-to-use products that do not require further optimization, or modification. Each reagent comes with a package insert that includes detailed instructions on its intended purpose. The following documents are provided upon request:

• Certificate of Analysis (CoA)

• Certificate of Conformance (CoC)

• Declaration of Conformity (DoC)

Many of our CE-IVD antibodies are available as recombinant REAfinity Antibodies, which are produced in a biologically and chemically defined in vitro expression system. This provides an additional level of standardization, leading to greater reproducibility of results.

Ready-to-go CE-IVD

REAfinity™ Recombinant Antibodies – optimized for flow cytometry

Our portfolio of recombinantly engineered antibodies provides great lot-to-lot consistency and purity.

The recombinant technology diminishes the need for FcR blocking and allows for analyses with only a single isotype control. This results in reproducible data with no more background signal, while saving efforts when setting up experiments.

For more information, visit: www.miltenyibiotec.com/reafinity

Product highlight

CD34/CD133 Enumeration Kit in CE IVD quality grade

CD34+ and CD34+/CD133+ cells harbor the capability to differentiate into cell types of several different tissues and are therefore interesting for various therapeutic uses. CD34 and CD133 enriched stem cell grafts have been employed in autologous and allogenic transplantation both in haploidentical¹-³ as well as HLA-matched settings⁴.

To provide a simple and standardized tool for the enumeration of CD34+/CD133+ cells by flow cytometry, we have developed the CD34/CD133 Enumeration Kit, based on the International Society of Hematotherapy and Graft Engineering (ISHAGE) guidelines. The kit is suitable for single-platform approaches, enabling absolute cell count determination by analysis of commercially available counting particles. A variety of human cell sources can be used, including whole blood, leukapheresis harvest, bone marrow, cord blood, and all fractions of CD34+ and CD133+ cells separated using the CliniMACS® System (fig. 1). More information on the CD34/CD133 Enumeration Kit (order no. 170-070-709) can be found in the respective package insert.

Literature: 1. Lang, P. et al. (2004) Br. J. Haematol. 124: 72–79.2. Lang, P. et al. (2004) Bone Marrow Transplant. 33: 879–880.3. Bitan, M. et al. (2005) Exp. Hematol. 33: 713–718.4. Bornhauser, M. et al. (2005) Leukemia 19: 161–165.

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A Bone marrow

Figure 1: Flow cytometric analysis of CD34+/CD133+ cells using the CD34/CD133 Enumeration Kit. Different starting materials, including bone marrow (A), cord blood (B), leukapheresis product (C), and CliniMACS CD34+CD133+ fraction from leukapheresis harvest were analyzed following the displayed gating strategy. For the bone marrow sample (A) all gating steps 1–9 are shown. For the other starting materials (B–D) steps 4, 7, and 9 are displayed. All samples were stained with the CD34/CD133 Staining Cocktail containing anti-CD45-FITC, anti-CD34-APC, and anti-CD133/2-PE antibodies. The kit allows for the identification of CD45+ leukocytes, CD34+ hematopoietic progenitor cells, and CD34+/CD133+ hematopoietic progenitor cells. Dead cells can be excluded from the analysis by adding 7-aminoactinomycin D (7-AAD), which is also included in the kit. The reproducibility CV was <0.1 for all specimen tested. Please refer to the package insert for sample requirements and preparation of solutions.

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2. Leukocytes 3. Viable cells CD34+ cells CD34+ CD133+ cells

CD133+ determination

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Gene-engineered T cells, such as chimeric antigen receptor (CAR) T cells, are T cells that have been redirected to elicit an anti-tumor immune response by targeting a specific tumor antigen. The detection of immune cell populations and determination of transduction efficiency by flow cytometry is important for the production process and can be used as part of the release criteria for the final CAR T cell product.

We have developed two panels of recombinantly engineered REAfinity Antibodies that can be used to detect immune cell populations and measure CAR T cell transduction efficiency by flow cytometry. Together with the corresponding MACSQuant Express Modes, our antibody panels provide a high level of standardization, facilitating reproducibility in flow cytometry applications.

Panel

FluorochromeImmune cell composition*

CAR T cell transduction* Clone

VioBlue CD45 CD45 REA747

VioGreen CD4 CD4 REA623

FITC CD3 CD3 REA613

PE CD16/CD56 CD19 or CD20 CAR Detection reagent**

REA423/REA196

PerCP 7-AAD** 7-AAD**

PE-Vio 770 CD20 or CD19 Free channel for additional marker of choice

REA780 or REA675

APC CD14 CD14 REA599

APC-Vio 770 CD8 CD8 REA734

MACSQuant Express Mode

Yes** Yes**

Antibody panels for immune phenotyping and detection of CD20+ and CD19+ CAR T cells

* Unless otherwise noted, reagents are available in CE-IVD quality. ** Available in CE-IVD quality soon.

Quality control in cell manufacturing

miltenyibiotec.com/IVD

Germany/Austria/ SwitzerlandMiltenyi Biotec GmbH Friedrich-Ebert-Straße 68 51429 Bergisch Gladbach Germany Phone +49 2204 8306-0 Fax +49 2204 85197 macs@miltenyibiotec.de

USA/CanadaMiltenyi Biotec Inc. 2303 Lindbergh Street Auburn, CA 95602, USA Phone 800 FOR MACS Phone +1 530 888 8871 Fax +1 877 591 1060 macs@miltenyibiotec.com

AustraliaMiltenyi Biotec Australia Pty. Ltd. Unit 16 A, 2 Eden Park Drive Macquarie Park NSW 2113 Australia Phone +61 2 8877 7400 Fax +61 2 9889 5044 macs@miltenyibiotec.com.au

BeneluxMiltenyi Biotec B.V. Sandifortdreef 17 2333 ZZ Leiden The Netherlands macs@miltenyibiotec.nlCustomer service The NetherlandsPhone 0800 4020120 Fax 0800 4020100Customer service BelgiumPhone 0800 94016 Fax 0800 99626Customer service LuxembourgPhone 800 24971 Fax 800 24984

ChinaMiltenyi Biotec Technology & Trading (Shanghai) Co., Ltd. Rooms 2303 and 2309 No. 319, Xianxia Road Changning District 200051 Shanghai, P.R. China Phone +86 21 62351005 Fax +86 21 62350953 macs@miltenyibiotec.com.cn

FranceMiltenyi Biotec SAS 10 rue Mercoeur 75011 Paris, France Phone +33 1 56 98 16 16 Fax +33 1 56 98 16 17 macs@miltenyibiotec.fr

ItalyMiltenyi Biotec S.r.l. Via Paolo Nanni Costa, 3040133 Bologna Italy Phone +39 051 6 460 411 Fax +39 051 6 460 499 macs@miltenyibiotec.it

JapanMiltenyi Biotec K.K. Nittsu-Eitai Building 5F 16-10 Fuyuki, Koto-ku, Tokyo 135-0041, Japan Phone +81 3 5646 8910 Fax +81 3 5646 8911 macs@miltenyibiotec.jp

Nordics and BalticsMiltenyi Biotec Norden AB Scheelevägen 17 223 70 Lund Sweden macs@miltenyibiotec.seCustomer service SwedenPhone 0200-111 800 Fax 046-280 72 99 Customer service DenmarkPhone 80 20 30 10 Fax +46 46 280 72 99 Customer service Norway, Finland, Iceland, and Baltic countriesPhone +46 46 280 72 80 Fax +46 46 280 72 99

SingaporeMiltenyi Biotec Asia Pacific Pte Ltd. 100 Beach Road #28-06 to 28-08 Shaw Tower Singapore 189702 Phone +65 6238 8183 Fax +65 6238 0302 macs@miltenyibiotec.com.sg

South KoreaMiltenyi Biotec Korea Co., Ltd Arigi Bldg. 8F 562 Nonhyeon-ro Gangnam-gu Seoul 06136, South Korea Phone +82 2 555 1988 Fax +82 2 555 8890 macs@miltenyibiotec.co.kr

SpainMiltenyi Biotec S.L. C/Luis Buñuel 2 Ciudad de la Imagen 28223 Pozuelo de Alarcón (Madrid) Spain Phone +34 91 512 12 90 Fax +34 91 512 12 91 macs@miltenyibiotec.es

United KingdomMiltenyi Biotec Ltd. Almac House, Church Lane Bisley, Surrey GU24 9DR, UK Phone +44 1483 799 800 Fax +44 1483 799 811 macs@miltenyibiotec.co.uk

www.miltenyibiotec.com

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121-

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Miltenyi Biotec provides products and services worldwide. Visit www.miltenyibiotec.com/local to find your nearest Miltenyi Biotec contact. Front cover image ‘Eye’ copyright © Miles Away Photography / Shutterstock.com.

CE-IVD reagents are for in vitro diagnostic use, and are not intended for human in vivo applications. In the EU, CE-IVD reagents are available as CE marked in vitro diagnostic medical devices and meet the requirements of the European directive 98/79/EC on in vitro diagnostic medical devices (IVDD). CE-IVD reagents are designed and manufactured under the ISO 13485 certified quality system. These products are not available for use in the United States. MACS and the MACS logo are registered trademarks or trademarks of Miltenyi Biotec GmbH and/or its affiliates in various countries worldwide. Copyright © 2018 Miltenyi Biotec GmbH and/or its affiliates. All rights reserved.