Welcome to a paradigm shift in biopharmaceutical downstream processing

Why a Paradigm Change?


CHRETO's single-use purification technology offers a paradigm shift for the biopharmaceutical industry by

 

  • Facilitating a fast, easy-to-use, scalable and cost efficient recovery of large biomolecules, in a single setup

  • Increasing productivity and reducing costs

  • Introducing the first truly single-use technology for the affinity purification of polyclonal (for plasma fractionation, IVIGs) and monoclonal antibodies (mAbs)


"The need for better single-use and disposable systems in downstream processing is growing…The move toward disposable upstream bioprocessing will push downstream operations toward innovative alternatives that may include membrane adsorbers and prepacked single-use chromatography columns."

 

"Disposable Chromatography: Options Are Increasing" Genetic Engineering News, Dec. 10, 2013

Learn how CHRETO Technology® works

DAP and CHRETO technology®

The principle of the CHRETO Technology® is straightforward
 The DAP molecules bind specifically to the target molecules in solution and form complexes. The complexes are separated on a support in the form of a membrane. After washing away the impurities, the complex is dissolved and the target molecule can then be collected.
 This procedure allows the process to be membrane based as well as single use which gives a long list of advantages over the current column technology based on Protein A, without compromising process yield and product quality. 

CHRETO Technology is a single-use, membrane-based affinity purification technology that will revolutionize downstream processing for the Biopharmaceutical industry. The technology is based on a DUAL AFFINITY PROTEIN (DAP) molecule which is constructed combining binding domains from protein A (alpha-helix structures in blue, below) that binds to the Fc region of antibodies and streptavidin (beta-sheet structures in yellow, below) that display high affinity for biotin (structure in red at the bottom of streptavidin). Reaction conditions in CHRETO Technology are radically different, much faster and more efficient than current column-based approaches. The figure below depicts a DAP monomer.

CHRETO Technology®

The proprietary CHRETO Technology® is a single-use kit format with a recombinant "Dual Affinity Protein" (DAP) molecule binding to the target molecule in solution, a generic membrane support, buffers, assays and ancillaries. The kit package format strongly supports the biopharmaceutical industries' need for manufacturing process simplicity, easy of use, fast set-up and linear scalability from R&D to pilot and full manufacturing scale. Advantages for the Plasma Fractionation Industry (IVIG production) CHRETO Technology® avoids the technical and practical shortcomings of the 60 year old Cohn fractionation technology and provides novel business opportunities due to improved yields, higher purity and better safety. The price competitive CHRETO Technology® will enable closing the gap between market demand and product availability of gamma-globulins. The plasma fractionation industry can significantly increase earnings when they implement CHRETO Technology® in their production. CHRETO® Technology improves the IVIG yield from plasma above 90 %. The yield will be up to 7.2g/litre of plasma depending on the source and fraction chosen. Advantages for the Biopharmaceutical Industry (mAb production) CHRETO Technology® avoids the technical and practical shortcomings of the 30 year old column affinity chromatography technology. Especially the bottleneck issues, packaging of large diameter columns with difficult-to-control flow properties as well as need for repeated extensively quality controlled cleaning and validation steps.
CHRETO Technology® process overview illustrating the catch, clean and collect steps. See details below (How does it work?).

How does it work?

The characteristics of the DAP molecule include 1) easy elution and no leakage, 2) specific binding to product (to secure selective reaction with target protein(s) and easy dissolving of formed DAP-target complexes and 3) tight binding to solid support (to secure immobilization and final removal of DAP molecules from product stream)

 

The purification process consists of three steps: catch, clean and collect.

  1. Catch. The DAP molecule reacts in solution with the feed stream containing the target molecule(s) e.g., an antibody.

  2. Clean. Impurities including HCP (host cell proteins) and DNA are removed followed by dissociation of the DAP-antibody complex and capture of DAP to a biotin support.

  3. Collect. The purified antibody is collected and the membrane cartridge is discharged.

Productivity enhancement

CHRETO Technology® provides cycle time improvements. Process savings per batch are 20 hours, corresponding to 40-50 days in manufacturing facility per year.
Comparison of Protein A column process versus CHRETO Technology®. Results were obtained at a workshop on manufacturing costs assessment in collaboration with a major biopharmaceutical manufacturer and NNE Pharmaplan, based on a process including 2000 L and harvest at 5 g/L.

Lab results

Below, two examples of the application of the technology are shown.

Plasma Fractionation CHRETO Technology® was used to purify gamma globulins from human plasma (IVIG). The recovery was 6 g of gamma globulin/L plasma achieved in our prototype test represents a 1.5-fold increase in yield compared to industry standard of 4 g/L plasma. Furthermore, the current CHRETO Technology® improves the IVIG yield from plasma above 90 %. The yield can reach 7.2g/litre of plasma depending on the source and fraction chosen.

The left-hand figure (above) shows the chromatogram profile, with three major protein peaks representing, from left to right: proteins not bound to DAP, proteins washed away from DAP-target complexes and the elution of the DAP-purified IVIG, respectively. Arrows indicate the fractions that are further analysed by SDS-PAGE (figure on the right). Lanes 6-8 on the SDS-PAGE contain highly purified gamma globulins.

 

High purity was obtained using a single purification step, as can be seen in lane 7-9 on the SDS-gel (right-hand panel above).

mAb purification CHRETO Technology® was used to purify a monoclonal antibody (mAb) using directly the clarified cell broth.

The left-hand figure (above) shows the chromatogram profile, with two major protein peaks representing the fractions containing the flow through after reaction with DAP (Fraction 7 and Lane 7 on the SDS gel, right panel above), and the fraction containing the purified mAb (Fraction 8 and Lane 8 on the SDS gel above). Lane 3 shows purified DAP; Lane 4 shows the feedstock used and Lane 6 show the same sample run without addition of DAP. Lanes 1 and 10 include the MW marker.

Company

Background
CHRETO was funded in 2010, as a spin out of Novozymes, by the inventor Jan Kyhse-Andersen and Torben Jørgensen to develop and commercialize a novel protein purification technology for bio-manufacturing.

 

CHRETO is based on a set of fundamental values

 

Creativity, Honesty, Respect and Trustworthiness

The company acquired in 2014 the rights to the original IP related to protein purification technology from Novozymes.


A prototype of the technology has been developed and thoroughly tested by industry experts and a leading pharmaceutical company in Europe during 2014-2016.

Vision and Goals

Our vision is that CHRETO Technology® will radically improve the process efficiency of downstream processing of large biomolecules. We believe that CHRETO Technology® will bring a true revolution to biomanufacturing, which has been sought by the industry for decades. 
Our goals are: 1) to have the first product evaluated in 2020 (achieved), 2) to launch the first commercial product in 2021 and 3) to position CHRETO as the preferred purification technology for IVIG and monoclonal antibodies going forward.

Funding

CHRETO has been supported by funding from an Eurostars project, a collaboration agreement with a major European pharmaceutical and Skeikampen Holding ApS together with National grants.

 

In September 2016, Biotage AB and Brohuvudet AB invested a total of DKK 20 million (15 + 5, respectively)  and Innovationsfonden invested DKK 4.6 million in CHRETO to secure development of the proprietary purification technology.

 

Shareholders (77 %)

 

  • Jan Kyhse-Andersen Holding ApS
  • Biotage AB
  • Novozymes A/S
  • Brohuvudet AB
Board of Directors

Who we are

Meet the the people working at CHRETO

Board of Directors

Kim Bjørnstrup - Chairman of the board

Kim  has a management carrier of more than 25 years in the pharmaceutical industry. He graduated  from Copenhagen University in 1984 and was licensed as a Lawyer in 1987.

From 1987 to 1992, he was Internal Legal Counsel to Lundbeck AS and Later Coloplast AS.

From 1992 to 2010 he worked at Octapharma AG in Switzerland, a swiss biotech, initially as Licensing manager and the last 10 years Vice Chairman /CEO. The Company had revenues exceeding one billion EURO and 4000 employees.

In 2010, he re-started Xeltis AG a Swiss biotech with a group of investors, specialising in self-generating heart valves.

From 2013 to 2014, he was the CEO of BPL UK a company with revenues of 300 million EURO and 2000 employees.

From 2015 he worked as CEO at Assistance HR Partners AS, where he I s co-owner, a company specialising in management training recruitment and temporary work.

 

Lars Bäckman - Member of the board

Lars Bäckman holds the position as VP of Corporate Development with Biotage AB since January 2007 and he is also the General Manager of Biotage subsidiary MIP Technologies. Prior joining Biotage, Lars Bäckman held the position as SVP of Corporate Development at Affibody. He has previous experiences acting as a lawyer at the Swedish law firm Hamilton & Co, specialized in Corporate Finance and Mergers & Acquisitions, and from Venture Management at the Business Development firm Googol. Mr. Bäckman holds a LLM from the Stockholm University and was born in 1961.

Sussi Wetterlin - Member of the board

Sussi is CEO of BASTAonline, a company focused on the substitution of chemicals of concern in the Building and Construction sector.

 

Sussi has more than 20 years of experience from the chemical industry within the environmental and commercial area. Sussi has extensive managerial experience with proven track record of leading international sales organisations during build-up, growth and turnaround phases. She held a position as European Sales Director in the Wood Industry sector with focus on the emerging markets in Eastern Europe and Russia.

 

Sussi has a M.Sc in Toxicology from Karolinska Institutet in Stockholm

 

Mario Augusto Maia - Member of the board

Business Development Novozymes A/S

Frank Nygaard - Member of the board

CMC and Project Director, Symphogen A/S

Click on the names to see CV

Contact us

Get in touch with us to hear more about DAP technology and its exciting potential for the biopharmaceutical and plasma fractionation industries.

Tel: +45 22 92 80 82

 Email: info@chreto.com

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CHRETO :: Lejrvej 17 :: DK-3500 Værløse :: Denmark :: tel: +45 22 92 80 82 :: Email: info@chreto.com