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고체상 접합기술을 이용한 항체-약물 복합체 (ADC)의 합성

Title
고체상 접합기술을 이용한 항체-약물 복합체 (ADC)의 합성
Other Titles
Application of solid-phase conjugation technology for synthesis of Antibody-Drug-Conjugate (ADC)
Author
김원준
Alternative Author(s)
WONJUN KIM
Advisor(s)
이은규
Issue Date
2018-02
Publisher
한양대학교
Degree
Master
Abstract
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.
URI
http://www.dcollection.net/handler/hanyang/000000105637http://repository.hanyang.ac.kr/handle/20.500.11754/68101
Appears in Collections:
GRADUATE SCHOOL[S](대학원) > BIONANOTECHNOLOGY(바이오나노학과) > Theses (Master)
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