Brazilian company develops and exports DNA polymerases

Fábio de Castro  |  FAPESP Innovative R&D – Cellco Biotec, a company headquartered in São Carlos, São Paulo State, has developed a novel process that enables high-quality DNA polymerases to be produced on a commercial scale for the first time in Brazil.

The project was supported by FAPESP’s Innovative Research in Small Business Program (PIPE), and was so successful that besides offering the Brazilian market enzymes with comparable quality to the imported product and at a more attractive price, the firm was able to sign its first export contract with a European client.

The researcher responsible for the project, Amanda Bernardes Muniz, explains that DNA polymerases are enzymes that are widely used for in vitro manipulation of DNA, including DNA cloning, sequencing and mutagenesis, among other techniques. Their applications range from medical diagnosis to forensic analysis based on genetic material, to cite just two examples.

“Although DNA polymerases are routinely used by laboratories, until now Brazil has never produced them to a sufficiently high standard of quality for commercialization. Our goal was to produce enzymes in Brazil with comparable quality to the imported products already on the market,” Muniz tells Innovative R&D.

Extensively trained in physics and molecular biology, Muniz says the firm developed quality control processes with the support of Jena Bioscience, a German company that is one of Cellco’s financial backers. “These enzymes are produced in bacteria, and are rendered useless if contaminated by any other DNA during the production process, so rigorous quality control is fundamental to commercialization,” she explains.

Maria Amélia Dotta, a biologist who worked in the pharmaceutical industry before becoming one of Cellco’s founders in 2014, says the need for import substitution was a key driver. “We were users of these enzymes and were deeply frustrated by having to depend on imports,” she says.

“Price fluctuations and local currency depreciation made it very hard indeed to plan projects and deliver results on schedule,” Dotta adds. “This experience in our work with molecular biology showed us that there was an opportunity to develop a product. We began doing so and decided to apply to FAPESP for funding.”

Dotta met Muniz while both were studying for a PhD at the University of São Paulo’s São Carlos Physics Institute (IFSC-USP), where they shared a research interest in recombinant proteins. Muniz joined the firm in 2016, and in the same year the project was approved by PIPE-FAPESP.

In 2018, having developed the production process, the firm won another grant from PIPE-FAPESP for Phase 2, which aimed at commercialization. “Even before the end of the project, the product was on the market, thanks to refinement of the enzyme developed in Amanda [Muniz]’s project,” Dotta recalls.

Muniz explains that genetic engineering enabled them to enhance the properties of the enzyme, which is Taq DNA polymerase (isolated from Thermus aquaticus), one of the most widely used and increasingly deployed in genotyping and diagnostics.

“By adding certain protein domains [independent parts of the polypeptide chain], we produced an enzyme that makes more processive polymerases. In other words, it’s faster than the normal Taq polymerase,” Muniz says.

According to Dotta, refinement of the product thanks to their research guaranteed an excellent market response. “We’ve developed a product of very high quality. This gives us the confidence to offer a product that’s been exhaustively studied and tested,” she says.

The presence of microbial DNA and inhibitors of the PCR reaction in the preparation process has to be avoided to guarantee the quality of the enzyme. This is one of the main challenges of commercial production and validation for industrial applications.

Cellco developed a quality control system that assures production of a high-performance and high-purity enzyme thanks to its partnership with Jena Bioscience, which now buys the product.

“As for all Cellco’s products, we developed the entire quality control process on the basis of Jena’s requirements for acquiring enzymes produced by other companies,” Muniz says. “All our processes are submitted to Jena’s very rigorous quality control, which gives us security.”

According to Dotta, Cellco has exported its enzyme to Germany since 2018 and has become Jena’s main supplier. Founded in 1998, Jena produces and supplies reagents to research laboratories and pharmaceutical companies in several countries.

“Jena invested in us from the start, possibly with the expectation that we would become a supplier of theirs. They saw Brazil as a promising market for life sciences, and especially for reagents,” Dotta says.

The Latin American market for DNA polymerases, according to the researchers, was estimated at BRL 1 billion in 2020, and is set to grow 6.7% per year until 2025, reaching BRL 1.38 billion.

Besides the two phases of the enzyme development project, completed in February 2021, Cellco also won support from PIPE-FAPESP for a project currently under way to develop a kit for diagnosing COVID-19 by multiplex RT-qPCR (reverse transcription quantitative polymerase chain reaction). The multiplex method, in which all reactions take place in a single tube, makes the process considerably cheaper (more at: https://agencia.fapesp.br/33891). 

In another project funded by PIPE-FAPESP, Cellco had already developed a process to produce deoxyribonucleotide triphosphates (dNTPs), used to amplify DNA in PCR testing with the help of Taq DNA polymerase.