고체상 접합기술을 이용한 항체-약물 복합체 (ADC)의 합성
- 고체상 접합기술을 이용한 항체-약물 복합체 (ADC)의 합성
- Other Titles
- Application of solid-phase conjugation technology for synthesis of Antibody-Drug-Conjugate (ADC)
- Alternative Author(s)
- WONJUN KIM
- Issue Date
- Antibody drug conjugate (ADC) is one of the topics for the next generation oncology therapeutics because of its higher therapeutic index and potency. ADC consists of three basic components: antibody, linker, and cytotoxic drug. Many research has tried to optimize the combination of these components through conventional solution-phase conjugation, which requires purified antibody for conjugating and also multi-steps of buffer exchange and purification after conjugation. Solid-phase conjugation, in which antibody is first immobilized to solid matrix by affinity interaction, reduced by a reducing agent to expose the reactive thiol groups, and then a drug-linker complex is specifically conjugated to the immobilized antibody through thiol-maleimide reaction followed by ADC elution from the matrix. Solid-phase conjugation could be a viable alternative since it would circumvent several post-conjugation purification steps, and eventually achieve a higher conjugation yield. Also, the immobilization step in solid-phase conjugation process could achieve purification and conjugation step simultaneously
i.e., it could synthesize ADC directly from antibody secreting cell culture broth. In this study, we used Herceptin® (antibody), SMCC (linker), and doxorubicin (drug) for ADC synthesis. We performed the solid-phase conjugation of ADC from purified antibody solution (“neat solution”) and culture broth mimicking solution. The results are identical indicating that the solid-phase conjugation process is not significantly interfered by the culture broth ingredients. We also confirmed the reproducibility of solid-phase conjugation process, and characterized the synthesized ADC by RP-HPLC, size exclusion chromatography (SEC), MALDI-TOF/TOF mass spectrometer, surface plasmon resonance (SPR), and cellular cytotoxicity test. Protein A-based solid-phase conjugation process shows ca. 20% higher yield than the conventional solution-phase conjugation process, which suggests solid-phase conjugation process is a feasible and attractive process for ADC synthesis directly from cell culture broth.
- Appears in Collections:
- GRADUATE SCHOOL[S](대학원) > BIONANOTECHNOLOGY(바이오나노학과) > Theses (Master)
- Files in This Item:
There are no files associated with this item.
- RIS (EndNote)
- XLS (Excel)