Ayoxxa Biosystems

Ayoxxa Biosystems (stylized in its logo as AYOXXA) is a biotechnology company founded in 2010 in Singapore, and headquartered in Germany.[5][6]

Ayoxxa Biosystems

Type	Private
Industry	Biotech
Founded	Singapore (2010), Germany (2012)
Founder	Andreas Schmidt, Dieter Trau
Headquarters	Cologne , Germany
Key people	Rodney Turner (CEO), Dieter Trau (CSO), Markus Zumbansen (CTO), Andreas Richter (CFO)
Products	Development of a Protein chip platform based on the Bead-based Arrays, named LUNARIS (TM) multiplexing technology
Production output	Biomarker Detection Kits
Services	Multiplexing and Readout Services, Assay Development Services
Owner	Andreas Schmidt, Dieter Trau, Wellington Partners, NRW.Bank, High-Tech Gründerfonds, Rainer Christine, Gregor Siebenkotten, KfW, b-to-v Partners, Creathor Venture, HR Ventures[1][2][3][4]
Number of employees	35
Subsidiaries	Ayoxxa Living Health Technologies Pte. Ltd (Singapore)
Website	www.ayoxxa.com

The company is known for developing protein chip capable of detecting at once multiple biomarkers, biomarker signatures (including markers for cancer, allergies, age related macular degeneration AMD, or infectious diseases) from a small biological sample. The protein chip yields large amounts of data, being primarily aimed for use in biomedical research in academia, clinic and industry.[5][7][8][9][10]

History

In 2006 the basic technology of the position-encoded bead-based Arrays multiplex protein chip began to be developed in the research labs of the Department of Bioengineering, at the National University of Singapore (NUS).[5][7][11] The project was led by Dieter Trau, Assistant Professor at NUS' Department of Bioengineering and Department of Chemical and Biomolecular Engineering.[5][7][12][13]

As a result, and with the continued support of NUS, the start-up Ayoxxa Living Health Technologies Pte. Ltd was founded in 2010.[5][9][11] The rights to the intellectual property developed at NUS were exclusively licensed to Ayoxxa, for the company to further develop.[5]

In 2012 the company expanded operations to Europe, establishing its headquarters in Cologne, Germany, as Ayoxxa Biosystems GmbH.[6] The company now develops partnerships, services, commercialization and further research from its headquarters.[8][14][15]

In December 2016, Ayoxxa Biosystems GmbH officially announced Mr. Rodney Turner as CEO and Dr. Markus Zumbansen as CTO.[16] Dr. Andreas Schmidt stepped down from CEO position in October 2016 and remains associated with AYOXXA as Board Member.

Ayoxxa protein chip I

Ayoxxa's protein chip or microarray technology enables the detection of large number of diseases through the protein analysis of a single droplet of blood or other bodily fluids.[5][7][9][17] This technology contrasts with previously established methods which were restricted to a single point testing, requiring considerable amounts of biological sample, and limiting the amount of analytes tested from each sample.[7][9][10][11][12]

Example of 96, 384 and 1536 well plates normally used for ELISA

Example of 37,500-probe DNA microarray with enlarged inset to show detail.
Ayoxxa's protein biochip technology is developed to achieve a comparable multiplex protein analysis (simultaneous analysis of multiple components in a single sample)

Instead of yielding one data point at a time (as is the case with classic ELISA), this new technology provides up to 10,000 data points,[18] with efficient labor input, using samples down to 3 microliters (µL) (range of 3 x 10⁻³ mL).[19][20] The multiplex technology approaches a level of analytical power (in throughput and accuracy) only previously seen in DNA sequencing arrays.[8][14]

The protein chip is made of silicon and is used to identify and qualify protein markers for cancer, allergies, cardiovascular or infectious diseases.[9] It simultaneously identifies multiple analytes, and the interactions between them. The technology is developed to run manually as well as fully automated, and giving more results quickly at high-throughput (yielding large amounts of data). The development of this technology is especially directed at the support biomedical research in academic and industry research,[11][12] with longer-term uses in pharmaceutical screening and preclinical diagnostics.[6][7][8][15][21]

Protein microarray

Microarrays are built of an 8 cm x 2 cm silicon base plate, or biochip, with dozens of separate 'wells' or containers located on top, each one offering a capacity for up to 20 uL used to incubate with reagents, blocking or washing solutions. Each of these macro-wells, contain thousands of micro-wells at the bottom, each measuring about 1/20th the diameter of a single strand of human hair.[12][20][22]

The wells on the chip are filled with antibodies (proteins) linked to microspheres to reside in the cavities.[9][11][12] Antibodies by nature are produced by immune cells. They 'bind' specifically to the antigens on the surface of viruses, bacteria, and diseased cells to clear and to prevent health damages to the body. Since antibodies are highly specific proteins, binding to specific antigens, they have been exploited to identify and measure protein biomarkers in immunoassays (e.g. ELISA).[22]

Since different antibodies 'bind' themselves to antigens, this affinity can be exploited for protein analytics by filling the wells with thousands of antibodies, and observing if and how much of a target antigen is present in a sample, or not. Ayoxxa's biochip can be used to identify thousands of different proteins in a single sample, including markers for diseases, where before numerous of the classic ELISA assays would be required to achieve the same results.[7][11][14]

Microscopic photograph of microbeads. These are polymer nanoparticles, typically 0.5 to 500 micrometres in diameter. In protein laboratory work, researchers attach different antibodies to different beads, keeping a record of what specific type of antibody each bead contains, and then track those beads in order to observe which ones react when exposed to a sample.

To observe the reaction of the different antibodies to the sample, the chip is placed under a standard laboratory fluorescent microscope with a digital camera, which snaps shots of it. It is then possible to electronically analyze and generate data of the different molecules and proteins present in the samples.[5][17]

In-situ Encoded Bead-based Arrays (IEBA) technology

The technology centers on the patented In-situ Encoded Bead-based Arrays (IEBA),[23][24] originally developed at the National University of Singapore (NUS), of which Ayoxxa was made the exclusive licensee.[5][7][12] Unlike other available bead-based microarrays, IEBA achieves greater multiplexing capability by recording the position of randomly distributed beads without the need for physical labels for bead identification, using instead the assignment of unique coordinates to each bead with an in-house software.[14][17][19][20]

These batches of different beads, each batch being an assay or capture site for a specific biomolecule, are applied sequentially creating a unique pattern for every well. The coordinates of each individual bead in each sequential batch is recorded in a large map/decoding data table that is provided to the user alongside the carrier (like a USB flashdrive). The assay itself follows the sandwich ELISA principle with a read out based on a fluorescent reporter introduced at the final step of the assay. Following imaging using fluorescence microscope technology on the reacted chip, the company's analysis software identifies the individual beads according to their unique signals, later decrypted with the company's software to deliver a customized report to the user.[17][20] This approach significantly reduces the complexity of downstream analysis while increasing the number of individual protein targets that can be analyzed in very low sample volumes.[14][17][19]

Open platform and interoperability

Ayoxxa has stated that its focus is in developing an open and adaptable IEBA platform, that can be easily incorporated into laboratories by being compatible with the existing laboratory technology that researchers are accustomed to using. Furthermore, IEBA is being developed to run with automated liquid handling systems used in high-throughput screening. It has also stated its aim at making the IEBA an open platform, adaptable to the diverse research needs.[19][20] Ayoxxa's CEO Andreas Schmidt compared the company's open platform philosophy as that of iTunes and iPhone, where many different apps (different biomarkers) run on the iPhone (Ayoxxa’s platform).[11]

The IBEAs can be used from low sample numbers up to the standard 384 well plate format, basically being a more advanced ELISA, keeping the current standards and protocols.[17][19][20] The arrays are designed to be readily adaptable to standard high throughput screening systems.[14][19] The IEBA technology can be used with existing bioassays protocols (particularly the bioassays on beads), and readers without the need to invest in a flow cytometer or other capital-intensive devices to read it out.[19][20]

Recognition

The company has been recognized for its scientific advancements and entrepreneurial drive.[12] Some awards the company received include:

• In 2008, proof of concept grant from the National Research Foundation.[25]
• In 2010, S$200,000 SMART Singapore–MIT alliance of Research and Technology Innovation Award.[9]
• In 2010, first prize in Best of Biotech, a global life sciences business plan competition in Austria.[5][11][12][25]
• In 2010, the company was included among the world's top 50 most promising startup companies by the US-based Kauffman Foundation, often described as the world's largest foundation devoted to entrepreneurship.[11]
• In 2011, Skolkovo Award in INSEAD's 21st Business Venture Competition.[9][26]
• In 2013, runner-up award in the Asian Entrepreneurship Award held in Japan.[21]
• In 2013, GründerChampions (Founder Champions) KfW North Rhine-Westphalia regional award, which rewards successful new "creative and sustainable business ideas that create social value", given during the German Founders and Entrepreneurs Days.[27]
• in 2014, S$100,000 Promising NUS Start-up Award. This was as part of the Innovation & Enterprise (I&E) Awards organized by the NUS Enterprise and The National University of Singapore Society (NUSS). The organizers stated that the winners were "the start-ups that we believe have the greatest potential to scale and make a real impact through their products and technologies."[28]

References

1. "AyoxxA raises EUR 2.6 million in series A". BioCentury Extra. Redwood City, California: BioCentury Publications, Inc. September 20, 2012. Archived from the original on 2014-04-13. Retrieved Apr 2, 2014.
2. "AyoxxA Biosystems Closes EUR2.6M Series A for Protein-Analysis Biochips". Dow Jones VentureWire. New York: Dow Jones & Company. September 20, 2012. Retrieved Apr 13, 2014.
3. "Ayoxxa gets funding, collaborates with NUS". BioSpectrum. Mumbai, India: Athena Information Solutions Pvt. Ltd. January 21, 2013. Archived from the original on April 4, 2013. Retrieved Apr 2, 2014.
4. "HTGF portfolio company AYOXXA Biosystems wins new investors to close a € 9,0m Series B round". Bonn, Germany: High-Tech Gründerfonds. July 10, 2014. Archived from the original on 2014-11-05. Retrieved Oct 29, 2014.
5. Tan, Amanda (August 8, 2012). "Protein chip to detect diseases faster; Ayoxxa's technology will also help scientists learn more about infections. Some 5,000 new tech startups are registered in Singapore annually, many of which are developing cuttingedge innovations to improve the lives of people around the world. In the last of a threepart series, two of these companies, Ayoxxa and YFind, are featured" (PDF). The Straits Times. Singapore. p. B17, Money Section. Archived (PDF) from the original on 2014-04-13. Retrieved Apr 2, 2014.
6. Chng, Grace (January 24, 2013). "Start Singapore". The Straits Times. Singapore. p. Money section.
7. Ramchandani, Nisha (Jan 22, 2013). "AyoxxA, NUS in joint research to further develop technology". The Business Times. Singapore. Retrieved Apr 2, 2014.
8. Moran, Nuala (September 26, 2012). "Wellington Pads Fund with $90.8M for Life Sciences". BioWorld International. Atlanta, Georgia: Thomson Reuters. 17 (39): 6.
9. Greve, Henrich (June 30, 2014). "Market Entry: Where to Start-Up?". Fountainebleau, France: INSEAD. Archived from the original on 2014-08-13. Retrieved Oct 29, 2014.
10. lim, Valerie (February 28, 2012). "The Ayoxxa Platform for Protein Multiplexing: A Novel Approach for Quantifying Multiple Proteins In Very Small Sample Volumes". Total Biopharma. Archived from the original on 2014-04-13. Retrieved Apr 12, 2014.
11. Basu, Radha (January 1, 2011). "Testing time for biotech scientist, A biochip that can detect hundreds of diseases" (PDF). The Straits Times. Singapore. p. D4. Archived (PDF) from the original on 2014-04-13. Retrieved Apr 2, 2014.
12. "Powerful disease detecting chip" (PDF). The Straits Times. Singapore. October 23, 2011. p. News section. Archived (PDF) from the original on 2014-04-14. Retrieved Apr 2, 2014.
13. "JN Medsys: Pioneers of '80% faster' digital PCRs". BioSpectrum. Mumbai, India: Athena Information Solutions Pvt. Ltd. Oct 24, 2013. Archived from the original on April 13, 2014. Retrieved Apr 2, 2014.
14. "Biochips firm AyoxxA gets $3.37 million funding". BioSpectrum Asia. India. September 21, 2012. Archived from the original on October 31, 2012. Retrieved Apr 13, 2014.
15. "Ayoxxa Raises $3.4M to Develop Protein Bead Array Platform". Genomeweb. September 20, 2012. Archived from the original on 2012-09-24. Retrieved Apr 13, 2014.
16. "Unternehmensregister". Unternehmensregister. Archived from the original (Web) on 24 December 2016. Retrieved 26 October 2016.
17. "Innovative Biochips für das Protein-Multiplexing" [Innovative biochips for protein multiplexing]. Laborwelt (in German). Germany: BIOCOM. 14 (2): 10. 2013. Archived from the original (PDF) on May 31, 2014. Retrieved Apr 13, 2014.
18. "AYOXXA BIOSYSTEMS GMBH". Zurich, Switzerland: German Biotech Database. Sep 10, 2014. Archived from the original on October 29, 2014. Retrieved Oct 29, 2014.
19. "Innovative biochips for protein multiplexing". BioSpectrum Asia. MM ACTIV: 17–18. October 2014.
20. Ashford, Molika (October 2, 2012). "Ayoxxa to Use $3.4M in Financing to Build Bead Chips for First Customers". Genomeweb BioArray News. Archived from the original on 2012-10-13. Retrieved Apr 2, 2014.
21. Chng, Grace (June 6, 2013). "Start S'pore". The Straits Times. Singapore. p. Money section. Archived from the original on April 13, 2014. Retrieved Apr 2, 2014.
22. Elshal, Mohamed F.; McCoy, J. Philip (2006). "Multiplex bead array assays: performance evaluation and comparison of sensitivity to ELISA". Methods. Elsevier. 38 (4): 317–323. doi:10.1016/j.ymeth.2005.11.010. PMC 1534009. PMID 16481199.
23. Ghosh, S.; Schmidt, A.; Trau, D. (2012). "Fast Bead Detection and Inexact Microarray Pattern Matching for In-situ Encoded Bead-based Array". Proceedings of the International Conference on Computer Vision Theory and Applications (VISAPP) -- Sponsored by Institute for Systems and Technologies of Information, Control and Communication (INSTICC). Portugal: SciTePress – Science and Technology Publications. 2: 5–14. doi:10.5220/0003826800050014. ISBN 978-989-8565-04-4.
24. US application 2010075865, Trau, Dieter; Liu, Wen-Tso & Johnson Ng, Kian Kok, "microarray system and a process for producing microarrays", published 2010-03-25, assigned to National University of Singapore
25. Tham, Irene (October 14, 2011). "Obstacles in the way of local tech startups; Poll shows hightech firms find it hard to get funding and patents". The Straits Times. Singapore. Archived from the original on October 6, 2013. Retrieved Apr 10, 2014.
26. "21st INSEAD Business Venture Competition". France: INSDEAD. 2011. Archived from the original on April 12, 2014. Retrieved Apr 12, 2014.
27. "GründerChampions 2013" [Founders Champions 2013] (in German). Germany: Deutsche Gründer- und Unternehmertage| Deutsche Gründer- und Unternehmertage. 2013. Archived from the original on March 31, 2014. Retrieved Apr 12, 2014.
28. "Record number of winners for the 2014 Innovation & Enterprise Awards" (Press release). Singapore: National University of Singapore. April 15, 2014. Archived from the original on 2014-05-13. Retrieved 2014-04-22.

External links

• Ayoxxa Biosystems on Twitter