Olerup News April 2015

NEWS April 2015

Introducing Real-Time PCR...

Olerup SSP AB introduces a new real-time PCR product line, QTYPE™, for the quantitative detection of HLA alleles. The initial product, for low-resolution HLA typing, is set for introduction in the second quarter of 2015. Read more in the article on page 4

Providing more than reagents A few words from Roswitha Keller, General Manager, Olerup GmbH. Page 2

Olerup SSP – Teaching an old dog new tricks What’s Now, What’s New, What’s Next?

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Capture-HLA™ – A unique Next Gen approach for HLA typing By incorporating traditional HLA loci as well as non-HLA markers, Conexio will launch a proprietary suite of NGS products.

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Increased exon coverage with new range of class II SBT Resolver™ kits By entering the real-time PCR market, Olerup not only provides innovative real-time PCR kits, we are also proud to present our own-design, real-time PCR machine, the ACCUTYPER™ Page 5

Updated class II sequencing products from Conexio Genomics.

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New release of Assign™ SBT 4.7 Introducing GSA analysis function and other new features.

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NEWS

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Providing more than reagents Olerup News shares ongoing events, topics and product updates with its readers and customers. Pioneering SSPtyping in the early 90’s, Olerup has grown over the years and new products have been added to our portfolio. The second edition of Olerup News gives you an insight into coming qPCR products and instrumentation from Olerup, new updates to the SBT product line, and the latest news on XM-ONE® and Gamma-Type™.

NEW PRODUCT LINE – INTRODUCTION OF OLERUP qPCR KITS

The innovative Olerup qPCR kits, QTYPE™, that many of our readers have been waiting for will shortly be available in Europe, the Middle East and Asia. In conjunction with the release of QTYPE™, we are also introducing ACCUTYPER™, a qPCR instrument for a seamless laboratory workflow in combination with QTYPE™. …..SBT TOO

Apart from the SSP and qPCR kits, Olerup also supplies SBT products for HLA-typing, and it may interest our readers to know that the world-renowned Stefan Morsch Foundation in Birkenfeld, Germany – one of the biggest registry labs on the planet – has recently decided to use the SBT Resolver™ product line in its routine typing procedures. The SBT Resolver™ product line, developed and manufactured by Conexio Genomics, is provided by Olerup together with the well-renowned sequencing software Assign™ SBT. As part of Conexio’s commitment to improving SBT Resolver™ products, we are pleased to announce a revamped set of class II SBT Resolver™ assays, which you can read more about in this edition.

WORLWIDE DISTRIBUTION

Where do you get our products from?

If your laboratory is in Germany, Austria or the BeNeLux countries, you may know that you can order directly from Olerup GmbH in Vienna. But even if your laboratory is situated elsewhere, Olerup GmbH has a very well-established network of highly competent distributors who specialize in supplying products for the HLA and Blood Bank market.

In total, Olerup has more than 25 distributors worldwide, many of whom have supplied customers with Olerup products for more than 15 years. Our distributors stay with us because they know from experience how good Olerup products are. They place great value in the fact that Olerup is the world’s number one SSP supplier, and we have reached and maintained this position by always supplying top-class, up-to-date products. The introduction of the new Olerup qPCR kits once again provides HLA and Blood Bank laboratories with products that will greatly facilitate their typing work

Roswitha Keller General Manager, Olerup GmbH

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NEWS

Capture-HLA™ - Next-Gen HLA typing that stands out from mainstream solutions Amongst Conexio’s latest developments, we are proud to present Capture-HLA™, an innovative alternative to PCR-based Next-Generation sequencing approaches. Capture-HLA™ utilizes capture probes with a proprietary design for multiple loci to isolate DNA fragments containing HLA-specific sequences.

This unique probe design enables the detection of all currently reported HLA alleles, as well as any new alleles to be discovered in the future.

Using capture probe technology for HLA typing with Next-Generation Sequencing technology offers many advantages:

Fragment genomic DNA

Repair fragments and ligate adapters

- Simple sample preparation due to testing all loci in a single tube.

- Easily updated with additional probes for other transplant-relevant loci, including Gamma Block SNPs. - Eliminates all PCR-associated errors and artifacts.

Enrichment of complete fragments

Capture-HLA™ probes isolate HLA specific fragments

Enrichment of captured fregments

Analyse in Assign MPS ™

- Compatible with all current popular Next-Gen sequencers.

Preliminary testing with Capture Probes has demonstrated high resolution HLA typing with identification of novel alleles that have arisen due to nucleotide substitutions.

Find out more about Capture-HLA™ at EFI: Abstract session 9 held on 28th April 15:30-17:00, Room 3.

Olerup and venues 2015 Olerup products and representatives can be seen throughout 2015 at the following meetings, conferences and exhibitions. We look forward meeting you.

MEET US IN 2015 EFI April 26-29, Geneva, Switzerland

ASHI Sep 28-Oct 2, Savannah, Georgia

ASHI Regional April 30 - May 2, San Antonio, TX, USA

DGI October 8-10, Lübeck, Germany

East-West Immunogenetics Conference June 4-6, Ljubljana, Slovenia

APHIA October 11-14, Bangkok, Thailand

ASHI Regional June 19-21, Philadelphia, PA, USA

Eurotransplant October 15-16, Netherlands

Birkenfelder Tage June 10-11, Birkenfeld, Germany

MEDICA November 16-19, Düsseldorf, Germany

BSHI September 9-10, Cambridge, UK

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NEWS

Olerup introduces product line for quick automated real-time HLA typing Olerup introduces QTYPE™, a new reagent product line for HLA typing with real-time PCR. The first QTYPE™ products to be released enable detection of HLA alleles at low/ intermediate resolution. These initial products are set to launch mid-2015 for research use only, followed by the launch of CE-marked and FDA 510(k) cleared products early 2016. Olerup SSP real-time PCR kits are developed for use with a range of popular real-time PCR instruments.

Moreover, Olerup will also provide ACCUTYPER™, a reliable and convenient qPCR instrument that, in combination with QTYPE™, makes the laboratory workflow even more seamless. For more information about this instrument, see next page.

“To develop a user-friendly yet advanced product line for more automated HLA typing has been the focus of our development team for some time. We are now proud to be able to commercialize our new concept”, says Anna Hedlund, Head of R&D at Olerup.

The first QTYPE™ products to be introduced on the market are: A-B-DR-DQ, 8 loci, 128 wells/test A-B-C-DRB-DQ, 9 loci, 192 wells/test A-B-C-DRB-DQ-DP, 11 loci, 384 wells/test

Hydrolysis Probes Olerup real-time PCR kits utilize the multiplexing capabilities of hydrolysis probes to increase the capacity for unique assays in each well. Using hydrolysis probes allows robust typing from low-input DNA concentrations, and removes the risk of misinterpreting primer dimers and other PCR artifacts seen in real-time techniques based on intercalating dyes. Due to their sequence-specific nature, hydrolysis probes also contribute towards increased specificity of the assay, potentially decreasing the number of ambiguities in the reported results.

orescent signal with each PCR cycle. This is done automatically on the real-time PCR instrument, eliminating the requirement for gel electrophoresis or other end-stage detection techniques. The instrument outputs a data file reporting the change in fluorescent signal in each cycle for each detection channel and each well of the PCR tray.

This data file is imported into the HLA interpretation and result-handling program. Interpretation is automatic and displays the most likely genotype; the user is then free to select how the results are reported. The software mirrors the laboratory workflow and guides the user to the correct result.

By combining the multiplexing power, sensitivity and specificity of hydrolysis probes with up to 384-well kits, Olerup provides a robust, easy-to-use HLA-typing product line with minimal ambiguities, whilst still retaining plenty of capacity to cope with future IMGT / HLA database updates.

Hydrolysis probes report the presence of PCR amplification as an increase in flu-

Fluorescence

HLA - positive IC

HLA - negative

Cycles

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Completing the workflow Generating reliable data with ACCUTYPER™…

As part of the real-time concept, Olerup provides the plate-based real-time PCR amplification and detection instrument ACCUTYPER™, a reliable and convenient qPCR instrument that smoothens laboratory workflow in combination with QTYPE™ assays.

ACCUTYPER™ comes with the option of blocks for 384 and 96 well plates, which can be changed in a matter of seconds. The instrument, which is front loaded and only 60 cm wide, is easy to place in the laboratory.

Its silver heating block ensures fast and uniform cycling while the five excitation and emission filter channel combinations ensure the flexibility that is needed for HLA typing, as well as for many other multi­plexed applications. The instrument also features pre-installed intuitive control software, making installation and run management particularly carefree. ...powered by SCORE™ The test workflow is simple; just add the DNA to the master mix and dispense it into the plate. Then seal the plate and put it into ACCUTYPER™. The instrument takes over from then on. All you need to do, once the run is over, is export the result to SCORE™.

Data handling and result interpretation is powered by the Olerup HLA result handling software SCORE™. When adding the real-time module to SCORE™, the software was also redesigned to enhance user-friendliness. The new version of SCORE™ is designed around a simplified workflow for both QTYPE™ and SSP tests; we believe it is a real benefit for our users to use the same software both for real-time typing as well as traditional SSP typing. The workflow in SCORE™ is split across four main screens: the patient and sample handling screen, the test screen, the analysis screen and the report screen.

After running your assay in the real-time instrument, the raw data file is imported into SCORE™. Interpretation is performed automatically and the most likely genotype is displayed. Users are then free to select

how the results are reported. The software mirrors the laboratory workflow and guides the user to the correct result. At Olerup, we believe that it is important for our highly knowledgeable users to, if desired, be able to evaluate any result in scientific depth. This remains an important feature of SCORE™ and is available in the redesigned program for QTYPE™ assays as well as for traditional SSPs.

“ACCUTYPER™ is a robust and easy-touse instrument. The QTYPE™ kits, ACCUTYPER™ instrument and SCORE interpretation software complete the ‘Olerup HLA in real-time’ concept in one integrated system”, says Ben Passey, Development Manager at Olerup.

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Gamma-Type™– Identification of Gamma block SNP mismatches that are associated with higher risk of GVHD Conventional HLA typing in hematopoietic stem cell transplantation (HSCT) includes loci at only 3 of the 4 major MHC genomic blocks, resulting in potential haplotype mismatching. Realizing that the Gamma block may be a key locus when predicting haplotype matching, Conexio Genomics has developed a powerful new addition to the HSCT matching arsenal.

Gamma-Type™ is a simple PCR-SSP for 25 Single Nucleotide Gene Polymorphisms (SNPs) within the Gamma block of the MHC. This block includes the C4, C2 and Bf genes.

Collectively, the SNPs provide a highly polymorphic locus that offers additional MHC information for predicting haplotype matching/mismatching

Accumulating data supports inclusion of Gamma block typing in HSCT donor selection Recent studies have shown that Gamma-Type™ mismatches are associated with severe acute GVHD. At the recent BMT tandem meeting in San Diego, Feb 11-15, 2015, Dr. Medhat Askar and colleagues at the Cleveland Clinic presented their initial findings with the Gamma-Type™ assay. Investigating 236 HSCT recipient/unrelated donor pairs transplanted at their center (20002010), Dr. Askar and colleagues concluded that mismatches of more than one of the 25 SNPs in the Gamma-Type™ assay were associated with higher frequency of acute GVHD.

Updated Class II SBT Resolver™ assays Olerup is a proud distributor of Conexio Genomics’ SBT Resolver™ product line, providing a flexible and high-resolution Sanger-based HLA typing solution to customers. As part of Conexio’s commitment to improving this product line, we take great pleasure in presenting a range of recently released class II updates: -

SBT Resolver™ DRB1: LG-PD5.2-7.

Includes bi-directional sequencing of exon 3 covering all DRB1 allele groups.

-

SBT Resolver™ DQB1: AN-PD6.2-3.

Full coverage of both exon 2 and exon 3 with bi-directional sequencing.

-

SBT Resolver™ DPB1: KD-PD10.2-1.

Complete coverage of exons 1-5.

NEWS

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New software release of Assign SBT 4.7 A new release version (v4.7.1) of Conexio’s fourth generation SBT analysis software is imminent. The new version will be available for download from: http://www.olerup-sbt.com/ assign/. New features include: Autosave feature A new autosave feature creates a backup copy of the user’s layout 5 minutes after a change is made. If you are working on a saved layout and the software closes unintentionally (due to PC error, software crash, etc.), the backup file remains and can be loaded so that unsaved changes can be reapplied to your project.

GSA analysis function This new function enables analysis of sequencing data produced using Group Specific Amplification (GSA) reagent solutions.

- Importing all GSA data at once and producing a single result/report dramatically streamlines and reduces analysis time for the user.

- Users can easily define their own GSA strategy in the software settings and choose whether to analyse against either genomic or cDNA references.

- Assign™ SBT 4.7.1 is the first fully flexible SBT analysis software that permits locus specific amplification, HARPS and GSA analysis.

- A high-resolution heterozygous typing result is produced by importing and analysing all GSA data for a single sample together. This includes analysing two alleles by two separate GSAs and/or multiple GSAs for a single allele if necessary.

For a complete list of changes from the previous version (Assign™ SBT v4.7.0.232), please refer to the Assign™ SBT v4.7.1 Update Document (TEC184).

Sequence Analysis Software

Conexio’s 4th Generation SBT analysis software

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Continuously updating the SSP portfolio With more than 20 years of experience in oligo design and SSP kit development, Olerup SSP AB continues to provide the most up-todate SSP HLA typing kits on the market. We take pride in being a leader in HLA typing with SSP technology. Today, we make valuable use of all our expertise in oligo design and the manufacturing of reliable HLA typing products by developing and introducing real-time PCR-based products, in addition to our strong SSP portfolio.

Nevertheless, our focus on designing and manufacturing top-of-the-line SSP products will also be maintained in the future.

Class II SSP tray to provide enhanced typing information catered towards common patient antibody/donor interactions.

In addition to producing SSP kits with superior quality and resolution, we are also dedicated to meeting new requirements and requests for product performance and design. Some of our most recent improvements include the DQA-DQB-DR Enhanced combi tray. In direct response to customer feedback, Olerup SSP AB recently launched this new

Enhanced Class II tray features • High-resolution DQA1 • DRB1, DRB345 and DQB1 low-resolution typing with the following enhancements (see Table 1 below).

Table 1 Resolution of null alleles

DRB4*01:03N and DRB5*01:08N

Resolution of common DR51 and DR52 groups DR51 groups: DRB5*01 vs. 02 DR52 groups: DRB3*01 vs. 02 vs. 03 Enhanced DRB1 resolution

DRB1*04: DRB1*04:01 vs. 04:02 vs.

04:03 vs. 04:05 vs. 04:07 DRB1*14: DRB1*14:01 vs. 14:54

Enhanced DQB1 resolution

DQB1*06: DQB1*06:01 vs. 06:02 vs.

06:03 vs. 06:04 vs. 06:09

What’s New During the first months of 2015, our team has been engaged in redesigning two of our Disease Associated kits as follows: Improved Olerup SSP® kit for alleles associated with Coeliac Disease • Redesigning the Olerup SSP® kit for alleles associated with Coeliac Disease, DQA1*02,05; DQB1*02,0302 (101.90324), has improved resolution focused on common, well-documented (CWD) alleles and facilitated the interpretation process.

These changes are further outlined in our new Tech Note on the subject, available through our website.

Improved Olerup SSP® kit for alleles associated with Narcolepsy • The redesigned Olerup SSP® kit for alleles associated with Narcolepsy, DQB1*06:02; DQA1*01:02 (101.901-24), follows the same focus of improved resolution regar­ ding common and well-documented (CWD) alleles. Note that due to customer demand, we have also included a DR*15group specific primer mix to the redesigned lot.

New Null allele kits • To meet the requirements of the 2015 NMDP policy for confirmatory typing of unrelated donors and patients, Olerup will expand its repertoire of convenient Add-on kits to include products for the additional null alleles that now require to be tested.

These new kits will be available during the first half of 2015.

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XM-ONE

XM-ONE

®

®

TRANSPLANTATION CROSSMATCH

A Tool for Enhanced Risk Assessment

Improving outcomes in kidney transplantation: Non-HLA antibodies and Enhanced Clinical Risk Assessment The XM-ONE® transplantation cross match is used prior kidney transplantation, to detect non-HLA antibodies against the donor’s endothelia. Results from the first clinical trial using XM-ONE®, published in February 2009 in the medical journal Transplantation1, demonstrated that XM-ONE® could predict rejection reactions in patients that were approved for transplants using test methods that are considered standard. Later publications have shown that XM-ONE® is a valuable complement to traditional antibody testing prior organ transplantation.

XM-ONE® is CE marked for sales in Europe and has been cleared by the FDA for sales in the U.S. Read more about XM-ONE® at www.absorber.se 1

Breimer, M, Rydberg, L, Jackson, AM et al, Multicenter evaluation of a novel endothelial cell crossmatch test in kidney transplantation, Transplantation 2009, 87(4):549-556.

Manufactured by:

Distributed by:

www.absorber.se

www.olerup.com

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Olerup @ the web Olerup web pages are easiest to access through www.olerup.com, the portal to our

For easy access, we have divided our currently available products into three broad category product web pages: SSP, SEQUENCING and ANTIBODY DETECTION. Information

distribution network. Among other information, you can find news about our companies, see what meetings Olerup will attend and, of course, get further information about our different products.

has been working with us for many years and are well-established partners to HLA laboratories in their respective region/country. Nevertheless, every year we add new distributors to our global network.

Distributor contact information is also easily accessible at the web page. Olerup is currently represented in more than 50 countries around the world. The majority of our distributors

about the new RT-PCR product line, which builds on the SSP-technique, can be found under the SSP pages.

As always, your feedback is important to us.

So if you have suggestions on how we can make our presence on the web even better, please let us know.

OLE-150412B-1

At Olerup, we strive to continuously develop and improve our web services. We believe that information and news of our products and companies should be easy accessible for users. If you have not recently visited our web pages, please do so and see how they can be of value for you.

PMS 2935 C

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Introducing QTYPE™ Speed and Precision in Real Time By combining the multiplexing power, sensitivity and specificity of hydrolysis probes, QTYPE™ is a quick, robust, and easy-to-use alternative in HLA typing. The technology ensures improved resolution whilst retaining plenty of capacity to cope with future allele updates.

As part of our real time concept, we also provide ACCUTYPER™, a reliable and convenient qPCR- instrument that will make the workflow with QTYPE™ seamless.

Data handling and result interpretation is of course powered by SCORE™

QT-150415 A-1

For Research Use Only.

Manufactured by:

Distributed by:

www.olerup-ssp.com

www.olerup.com