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Biotechnology and Life Sciences in Baden-Württemberg

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12.03.2012

Q-bios provides client-specific biotechnology services

Q-bios GmbH, a spin-off from the Mannheim University of Applied Sciences, is a biotechnology company that provides services related to the production and purification of diagnostic and therapeutic proteins as well as the development and amplification of suitable cell lines. The company offers its services, which are based on state-of-the-art technologies, to clients from the biotechnology, diagnostic and pharmaceutical industries.

Dr. Heiko Flammann (© private)

“Q-bios GmbH biotechnology" is an emerging biotechnology company based in Mannheim, offering services and products for the pharmaceutical, diagnostic and biotechnology industries. The company is particularly focused on the production of recombinant proteins and polypeptides as well as the development and amplification of cell lines according to client requirements. Q-bios uses a broad range of technologies from the fields of microbiology, molecular genetics, cell and immunobiology and protein analytics and can rely on the skills and long-term research experience of its founders, Dr. Heiko Flammann and PD Dr. Andreas Lux.

Economic exploitation of research know-how

PD Dr. Andreas Lux (© private)

Q-bios is a spin-off from the Mannheim University of Applied Sciences where Dr. Flammann and Dr. Lux worked as scientists in the Institute of Molecular and Cell Biology. According to Professor Dr. Mathias Hafner, head of the institute and mentor to the two company founders, creative start-up companies are best placed to be able to dynamically transfer research know-how into industrial applications when they have a team with scientific experience and competent external advisors. The establishment of Q-bios is proof of the institute’s efforts to turn biotechnological research results into the rapid and inexpensive production of biomaterials.

In 2009, Flammann and Lux won an EXIST business start-up grant from the German Federal Ministry of Economics and Technology that enabled them to turn their business idea into reality. Mannheim University of Applied Sciences provided the founders with rooms (including an S2 laboratory) and state-of-the-art equipment at the Institute of Molecular and Cell Biology and hence with the conditions required for the operative establishment of the company during the start-up phase.

Superior expression systems for the production of proteins

Mannheim University of Applied Sciences campus; headquarters of Q-bios GmbH (© Mannheim University of Applied Sciences)

The company uses different expression systems for the production of proteins of interest for diagnostic and therapeutic applications. The expression systems include bacteria, yeasts, insect cells such as Sf9 and Sf21 (suspension cultures in serum-free medium derived from the ovaries of the butterfly Spodoptera frugiperda which can be transfected with baculoviruses carrying the gene required) and mammalian cell cultures such as Chinese hamster ovary cells (CHO-K1), human embryonic kidney cells (HEK 293) and mouse myeloma cells (NSO). The company develops the cell lines and isolates individual clones according to client requirements based on client-specific cells. Q-bios has also developed a proprietary “Expression Enhancer System” (EES) which enables high quantities of proteins to be produced in transient suspension cell lines. This expression system is superior to other expression systems on the market. EES is also suitable for producing high amounts of proteins in stable suspension cell lines. Q-bios also offers modified and apoptosis-resistant HEK293 and NSO suspension cell lines for the transient and stable production of proteins.

The coding sequence of the sought-after protein will be cloned – tagged if necessary with histidine (His6) or glutathione-S transferase (GST) – into a cell-system-specific expression vector. The gene will subsequently be sequenced to verify whether cloning has been successful. The expressed proteins are purified using permeation, ion exchange and affinity chromatography using GST, protein A or protein G columns, or using nickel chelate chromatography.

The purity and quality of the clones are checked using gel electrophoresis and immunoblots involving tag- or protein-specific antibodies, or ELISA tests, if necessary. The company also offers immunofluorescence tests, polymerase chain reaction (PCR, including quantitative PCR) and different cell culture assays with all tests tailored to client requirements.

PCR enzymes

A Q-bios laboratory at Mannheim University of Applied Sciences (© Mannheim University of Applied Sciences)

Q-bios’ product line is mainly focused on the production and client-specific formulation of DNA polymerases. DNA polymerases such as those isolated from the thermostable bacterium Thermus aquaticus (Taq) are enzymes that are able to amplify the smallest quantities of DNA in the presence of suitable primer sequences, thus making the DNA detectable. The use of Taq polymerase has made polymerase chain reactions (PCR, with reaction temperatures of between 55 and 72^oC) applicable to a broad range of molecular biology and diagnostics problems, including the detection of bacterial and viral pathogens.

Q-bios can offer DNA polymerases with variable activities and formulations. For example, the Taq polymerase is often offered in combination with so-called “HotStart” mixtures in which specific inhibitors (e.g. antibodies, aptamers) prevent unspecific syntheses from occurring at low temperatures (e.g. at room temperature). This is done by inhibiting the polymerase’s activity at ambient temperature, and instantly activating it at the required reaction temperatures (e.g. above 50^oC).

Virus experts

Q-bios plans to expand its scientific knowledge and experience in handling adenoviruses and adeno-associated viruses (AAV) and make this area another business priority in the German market. AAV are single-strand DNA viruses that can be integrated into the genome of host cells, but are not replicated. In order for AAV to replicate in the host cell, an adenovirus needs to be present as this helper virus encodes the proteins that AAV need for replication. As AAV are not known to cause disease, but are excellent tools for infecting cells, they are not only used in research, but also increasingly for gene therapy. The production and purification of AAV and the production of recombinant AAV variants is a rather sophisticated service, which the company provides to research institutions that lack the necessary know-how.

The expertise of the Mannheim-based company in the field of virology might be put to good use in the diagnosis of hantaviruses (see yellow box). Some hantaviruses have been associated with human disease, some are known to cause potentially fatal diseases in humans, including haemorrhagic fever with renal syndrome (HFRS). Hantavirus diseases need to be notified to health authorities (ed. note: in Germany, to the Robert Koch Institute). The severity of disease depends on the type of virus that causes it. While the scientific typing of this virus group is established, there is still huge need for test methods that can reliably diagnose and differentiate the different types of hantavirus. And Q-bios also has something to offer for this problem. The Q-bios founders offer new, improved antigens for the development of reliable test methods for the medically most important types of hantavirus. A round-robin test has shown that Q-bios antigens can be successfully used in ELISA (enzyme-linked immunosorbent assay) tests to identify the virus type with which a patient has been infected.

Background: hantaviruses

Hantaviruses are enveloped, negative sense RNA [ss(-)RNA] viruses in the Bunyaviridae family. The name hantavirus is derived from the Hantan River area in South Korea where thousands of American soldiers developed haemorrhagic fever with renal syndrome during the Korean War at the beginning of the 1950s. The virus that caused the disease wasn’t isolated until 1977. Around 25 or so different hantaviruses have been described, of which around 50 % are associated with human disease (lung and kidney diseases, severe fever attacks, outer and inner bleeding). Hantaviruses occur worldwide. Hantavirus infections that cause a disease known as haemorrhagic fever with renal syndrome are also quite frequent in Germany. Typical disease symptoms are high fever, head- and muscle aches, skin and mucosal bleeding. Complications include renal failure of variable severity; hantavirus infection can get worse quickly, acute kidney failure can occur and may lead to death.

The most important types of hantavirus found in Germany are the following: Hantavirus, Puumala virus and the Dobrava-Belgrade virus. The Puumala virus is the main hantavirus species in Germany and induces milder forms of disease without haemorrhage. Incidence rates in different countries fluctuate considerably from year to year. Normally, only around 100 to 200 hantavirus infections are registered in Germany every year. However, the Robert Koch Institute registered as many as 1687 cases in 2007. In 2010, more than 800 hantavirus infections occurred in the German state of Baden-Württemberg alone. Transmission occurs mainly by inhalation of aerosolized droplets of secretions from infected rodents or aerosolized particles of dust etc. carrying the virus. In Germany, the bank vole Myodes glareolus seems to be the key source of infections; it has been found that human infections correlate with the occurrence of M. glareolus, which are occasionally found to reproduce quickly and massively.

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