Tell us about your project!
Our main project is to develop future cancer drugs, and we have developed a platform around antibody drug conjugates (ADCs), where we test various actionable receptors as novel ADC targets. This project uses the GlyCLICK kit to conjugate antibodies to highly potent linker-toxins (payloads). By using GlyCLICK, we can develop highly homogeneous ADCs with a well-defined drug-to-antibody ratio (DAR). Usually, ADC development relies on conjugation to the cysteine- or lysine-residues of an antibody, which result in highly heterogeneous products with varying DARs.
How did SmartEnzymes enhance your project?
When comparing the conjugation methods using MALDI-TOF mass spectrometry, the GlyCLICK prepared ADCs result in a narrow peak. In contrast, the cysteine-conjugated ADCs have several and broader peaks, suggesting that the GlyCLICK ADCs are highly homogeneous compared to the cysteine-conjugated ADCs. Even though techniques such as hydrophobic interaction chromatography (HIC) or reverse-phase HPLC can ensure more homogenous ADC batches, these methods are time-consuming, require expensive equipment, and generate lower ADC yield. In our project, drug homogeneity is of the highest importance, making the GlyCLICK preferred to other conjugation methods.
As GlyCLICK prepares the antibody for click-chemistry conjugation, the antibody can be conjugated to other components such as fluorescent dyes. This allows us to investigate how the conjugation method affects antibody functions such as binding affinity, stability, and internalization in target positive cell lines. Compared to interchain cysteine conjugation, we obtain more stable ADCs with the GlyCLICK technology since the cysteine conjugation relies on breaking some of the stabilizing cysteine bonds in the antibody.
Overall, the GlyCLICK kit has dramatically improved our project by making the ADC development more efficient and making the ADCs safer to use without losing potency.
Discover the SmartEnzymes used in this project!
GlyCLICK