Highlights

In brief

Targeted gene integration helps create a wider range of antibodies with diverse N-glycan structures to explore for new therapeutic options.

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Hitting the antibody manufacturing sweet spot

31 Oct 2022

Scientists create a platform for reliably customising antibody molecular structures, opening the door to more potent and effective antibody medicines.

Antibodies aren’t just the cornerstone of the human immune response: scientists have used them to create medicines against many diseases such as cancer and COVID-19, thanks to their intricate molecular structures can be customised to enhance or tailor therapeutic effects. For example, studding the surface of antibodies with chains of sugars called N-glycans can make antibody treatments more potent, or help them stick around in the patient longer.

However, mass-producing antibody medicines is no easy feat, given that they must be created in living cells. Currently, the pharmaceutical industry uses specialised cell cultures to manufacture antibodies, which experts say are unreliable when it comes to making the antibodies’ N-glycan ‘coats’.

“To improve N-glycan processing in these cells, genes encoding for glycan-modifying enzymes are traditionally overexpressed via random integration technology,” explained Yuan Sheng Yang, Principal Scientist at A*STAR’s Bioprocessing Technology Institute (BTI). This technique results in highly diverse batches of antibodies which can lead to inconsistencies in how these treatments work in patients.

Seeking to optimise this process, Yang led a group of researchers who developed a targeted gene integration technology. This novel platform uses an enzyme called recombinase which snips two pre-determined sites in the genomes of antibody-expressing cells, before swapping this section out for a specific gene sequence.

Yang’s team used targeted integration as a tool to study the mechanisms by which sugars are attached to antibodies during production. They studied a panel of 42 human genes linked to sugar synthesis, sugar transport and N-glycan chain extension.

Their experiments revealed that turning on a gene called B4GalT1 helped produce antibodies with thick coatings of galactosyl, an N-glycan known to boost the therapeutic function of antibodies. Moreover, the team showed that increasing the expression of a second gene, ST6Gal1, produced highly-sialylated antibodies, which prevents antibodies from being cleared from the body too quickly.

“Targeted integration technology can produce antibodies carrying a diverse range of glycan structures to suit different therapeutic needs,” Yang said, adding that the team still needs to figure out how to boost antibody production yields using the new technique.

“We hope to further improve the current platform to reach the production titre of industry standards for clinical or commercial use,” Yang commented. Future studies will also dive deeper into how various glycan profiles affect the function of therapeutic antibodies.

The A*STAR-affiliated researchers contributing to this research are from the Bioprocessing Technology Institute (BTI).

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References

Nguyen, N.T.B., Lin, J., Tay, S.J., M., Yeo, J. et al. Multiplexed engineering glycosyltransferase genes in CHO cells via targeted integration for producing antibodies with diverse complex‑type N‑glycans, Scientific Reports 11:12969 (2021) | article

About the Researchers

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Yuan Sheng Yang

Scientist, Group Leader of Animal Cell Technology Group 1

Bioprocessing Technology Institute (BTI)
Yuan Sheng Yang received his doctorate degree from the Department of Chemical Engineering at Vanderbilt University in 2005. Upon graduation, he joined A*STAR's Bioprocessing Technology Institute (BTI). Yang is currently heading the Cell Line Development group and with a focus on the development of platform technologies for accelerating the process of cell line development and antibody development. He has published 54 peer reviewed papers and has filed six patents. His cell line development and antibody development platform technologies have been licensed to dozens of companies, resulting in license revenues exceeding US$3M. The CHO cell line platform Yang helped developed in collaboration with industry partners features unrivalled productivity and speed, enabling the development of five antibody drugs into clinical trials.
Ngan Tran Bich Nguyen received her PhD in Molecular and Cell Biology from the National University of Singapore in 2018. Her graduate work focused on understanding the mechanisms of inflammation suppression and resolution to develop therapeutics for inflammatory lung diseases. She then joined the Cell Line Development group at A*STAR's Bioprocessing Technology Institute (BTI), where she is working on developing targeted integration technologies for CHO cell engineering to produce therapeutic antibodies with improved efficacy.

This article was made for A*STAR Research by Wildtype Media Group