Development of cell assays for exploration of novel immunotherapeutic drugs for cancer treatment
(2025) KIMM05 20251Department of Immunotechnology
- Abstract
- Bladder cancer remains a prevalent and deadly cancer form worldwide. While immunotherapy has yielded significant progress, additional therapeutic strategies are needed to improve the response in bladder cancer patients. A novel immune cell target, referred to as “protein X”, has previously been identified in Lundberg’s lab. Antibodies developed against protein X were provided to us, with the aim of establishing a set of cell-based assays for characterization and functional screening of the IgG candidates.
Using an IgG cell binding assay based on a cell line expressing human protein X, as well as on genetically modified HEK293 cells expressing murine or cynomolgus orthologues of protein X, we identified cross-reactive IgG clones. To... (More) - Bladder cancer remains a prevalent and deadly cancer form worldwide. While immunotherapy has yielded significant progress, additional therapeutic strategies are needed to improve the response in bladder cancer patients. A novel immune cell target, referred to as “protein X”, has previously been identified in Lundberg’s lab. Antibodies developed against protein X were provided to us, with the aim of establishing a set of cell-based assays for characterization and functional screening of the IgG candidates.
Using an IgG cell binding assay based on a cell line expressing human protein X, as well as on genetically modified HEK293 cells expressing murine or cynomolgus orthologues of protein X, we identified cross-reactive IgG clones. To evaluate off-target binding, peripheral blood mononuclear cells from healthy donors were used, enabling the selection of clones with low off-target interactions and promising developability. A neutrophil-based off-target assay, monitoring production of reactive oxygen species, confirmed that the IgGs did not induce stronger activation than nivolumab, an approved immunotherapeutic antibody. Finally, we evaluated various setups for a T cell proliferation assay and identified key parameters for the establishment of a robust screening. When optimized, this assay will be used to assess the immunosuppressive function of protein X.
Together, these screening approaches allowed initial screening of IgGs targeting protein X and identification of IgG clones with promising properties. Although further assay refinement and in-depth characterization is necessary, our results support the continued development of protein X-targeting antibodies and provide a foundation for their future use in immunotherapy against bladder cancer. (Less) - Popular Abstract
- Development of methods to find new antibody drugs against cancer
Bladder cancer is a common and serious disease that affects many lives around the world. While new immunotherapies – treatments that help the body’s immune system to eliminate tumor cells – have revolutionized bladder cancer treatment over the last decade, many patients still don’t respond to them. In this project, we took part in a larger initiative to identify new immunotherapies that boost the immune system’s ability to fight cancer.
One kind of immunotherapy is based on antibodies, which are biological molecules that selectively bind to proteins in our body to either block or enhance their biological functions. When developing antibody drugs, you commonly start... (More) - Development of methods to find new antibody drugs against cancer
Bladder cancer is a common and serious disease that affects many lives around the world. While new immunotherapies – treatments that help the body’s immune system to eliminate tumor cells – have revolutionized bladder cancer treatment over the last decade, many patients still don’t respond to them. In this project, we took part in a larger initiative to identify new immunotherapies that boost the immune system’s ability to fight cancer.
One kind of immunotherapy is based on antibodies, which are biological molecules that selectively bind to proteins in our body to either block or enhance their biological functions. When developing antibody drugs, you commonly start with a pool of candidate antibodies that are evaluated to find the best one.
In this study, we investigated a new promising immune target previously identified by Lundberg’s lab. Special antibodies that recognize this protein target were provided and our goal was to develop laboratory methods on living cells to evaluate the antibody candidates’ potential as immunotherapeutic drugs.
We first identified which antibodies could work across different species, an important step when preparing for future animal studies. To accomplish this, cells that carried human, mouse or monkey versions of the immune target were used. Moreover, we investigated whether the antibody candidates unintentionally bind to other cells in the human body, by testing them on human blood. Most candidates showed low stickiness, meaning they’re likely to be safe to use.
Secondly, we developed a method to investigate if the antibodies caused unwanted immune responses involving neutrophils – immune cells taking part in our first line of defense. None of the candidates triggered stronger responses than an already-approved cancer drug, suggesting sufficient safety potential. However, the tests showed that neutrophils can react very differently in people, and therefore the method needs more work to be reproducible and reliable. Lastly, we began to establish another method, this time using T cells – immune cells that are crucial for the defense against cancer. The goal was to investigate if the immune target indeed would suppress T cells, which it did! We also identified key protocol adjustments; however, more fine-tuning is needed for a successful method.
To conclude, this project contributes to the growing body of work aimed at identifying the top antibody candidates for new bladder cancer treatment. With continued improvement of the laboratory methods developed here, we are optimistic that these efforts in the end will provide a new drug candidate for bladder cancer treatment. (Less)
Please use this url to cite or link to this publication:
http://lup.lub.lu.se/student-papers/record/9201341
- author
- Fond, Fabian LU and Karlsson, Johanna LU
- supervisor
- organization
- course
- KIMM05 20251
- year
- 2025
- type
- H2 - Master's Degree (Two Years)
- subject
- keywords
- Bladder cancer, cell assays, antibody, immunotherapy, immuno-oncology, optimization
- language
- English
- id
- 9201341
- date added to LUP
- 2025-06-18 09:54:47
- date last changed
- 2025-06-18 09:54:47
@misc{9201341,
abstract = {{Bladder cancer remains a prevalent and deadly cancer form worldwide. While immunotherapy has yielded significant progress, additional therapeutic strategies are needed to improve the response in bladder cancer patients. A novel immune cell target, referred to as “protein X”, has previously been identified in Lundberg’s lab. Antibodies developed against protein X were provided to us, with the aim of establishing a set of cell-based assays for characterization and functional screening of the IgG candidates.
Using an IgG cell binding assay based on a cell line expressing human protein X, as well as on genetically modified HEK293 cells expressing murine or cynomolgus orthologues of protein X, we identified cross-reactive IgG clones. To evaluate off-target binding, peripheral blood mononuclear cells from healthy donors were used, enabling the selection of clones with low off-target interactions and promising developability. A neutrophil-based off-target assay, monitoring production of reactive oxygen species, confirmed that the IgGs did not induce stronger activation than nivolumab, an approved immunotherapeutic antibody. Finally, we evaluated various setups for a T cell proliferation assay and identified key parameters for the establishment of a robust screening. When optimized, this assay will be used to assess the immunosuppressive function of protein X.
Together, these screening approaches allowed initial screening of IgGs targeting protein X and identification of IgG clones with promising properties. Although further assay refinement and in-depth characterization is necessary, our results support the continued development of protein X-targeting antibodies and provide a foundation for their future use in immunotherapy against bladder cancer.}},
author = {{Fond, Fabian and Karlsson, Johanna}},
language = {{eng}},
note = {{Student Paper}},
title = {{Development of cell assays for exploration of novel immunotherapeutic drugs for cancer treatment}},
year = {{2025}},
}