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Development of one-step purification process to analyze the quality of the culture media
- Author(s)
- 이자연
- Advisor
- 서민덕
- Department
- 일반대학원 분자과학기술학과
- Publisher
- The Graduate School, Ajou University
- Publication Year
- 2020-08
- Language
- eng
- Keyword
- onestep; process; purification
- Abstract
- 본 "One-step" 연구는 현재 생산 중인 recombinant glycoprotein 배양액에서 품질분석을 위한 기존 다수의 정제 공정을 대신하는 단백질과 coupled affinity 정제를 이용하여 개발을 진행하였다. 여기서는 antibody-antigen 상호작용을 응용하여 recombinant glycoprotein과 based resin과 coupled 하여 정제 공정을 진행하였고 recombinant glycoprotein에 glycan이 다수 관찰되는 것을 이용한 glycan-targeting 레진들 (Immobilized D-galactose gel, HA ultragel, Con-A sepharose FF)를 사용하였다. 이 공정을 통해 기존 공정보다 좋은 performance를 기대하였다. 단백질과 resin과의 결합은 확인되었다. 이와는 대조적으로, 기존의 알려진 정제 공정과 다르게 affinity test에서 recombinant glycoprotein 이 낮은 affinity를 확인하였다. 다른 glycan-targeting resin은 단백질 구조에서 D-galactose가 immobilized D-galactose gel에 검출되지 않았고, HA ultragel과 Con-A sepharose FF 4B resin들은 단백질과 ligand 상호작용의 capacity가 낮았다. 따라서 커플링 공정으로 항체화한 단백질과 결합한 resin과 glycan-targeting하는 4가지 resin을 이용하여 recombinant glycoprotein에 대한 정제 공정을 기존의 정제 공정과 비교하였다.
- Alternative Abstract
- This "One-step" study is trying to develop a protein coupled affinity purification which is instead of a number of conventional processes for the quality analysis of items in the recombinant glycoprotein culture fluid currently in production. Here, We purification processes by coupled with based resin through recombinant glycoprotein antibodies applying antibody-antigen- interactions and observing for many glycan to recombinant glycoprotein of using glycan-targeting resins (Immobilized D-galactose gel, HA ultragel, Con-A sepharose FF) used it. We estimated the process that has good performance by comparing with the existing process through the process. The coupling process to based resin with protein was confirmed. In contrast, to its confirmed coupling resin use purification process did showed a low affinity in the purification process of recombinant glycoprotein that was other affinity tested. The other glycan-targeting resins are Immobilized D-galactose gel without to detected D-galactose in protein structures and the HA ultragel or Con-A sepharose FF 4B resins were low capacity ligand interactions with proteins. Thus, The purification process for recombinant glycoprotein was compared with the traditional purification process using three resins that are glycan-targeting and coupled with recombinant glycoprotein antibody resin.
- Fulltext
- Appears in Collections:
- Graduate School of Ajou University > Department of Molecular Science and Technology > 3. Theses(Master)
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