Production and engineering of recombinant antibodies
Korea (KMSA) - Ajou university, Gyung gi province
Department of Microbiology, College of Medicine
Myung-Hee Kwon
Myung-Hee Kwon
English, Korean
4 weeks
Cities/Months Jan Feb Mar Apr May Jun Jul Augt Sep Oct Nov Dec
No No No No No Yes Yes No No No No No
Type of Research Project
- Basic science
What is the background of the project?
The use of antibodies to modulate the functions of antigens by interacting with them inside cells in living cells is a valuable application in biotechnology, medicine, and cell biology. The rate by which a antibody is being internalized by cells is an important factor for successful modulation of a target protein function. Many techniques for antibody internalization including radiolabels, fluorescent microscopy, flow cytometry and cellular toxicity assays have been developed. However, none of these methods have yet evaded the necessary for purifying antibody protein to be examined and using a fluorescence-labeled secondary antibody and antibody modifications by genetic manipulation and/or chemical conjugation. Thus, a rapid and simple technique is need for the quantitative detection of cellular internalizing antibody.
What is the aim of the project?
Develpment of plate-based techniques for the quantitative detection of cellular internalizing antibody
What techniques and methods are used?
 For antibody production: gene cloning, cell culture  For antibody purification: affinity chromatography  For analysis of antibody activity: enzyme-linked immunosorbent assay (ELISA)  For analysis of antibody internalizaton: flow cytometry, fluorescence resonance energy transfer (FRET) assay, confocal microscopy, western blotting
What is the role of the student?
- The student will mainly observe
- The student will observe the practical experiments but will be highly involved in the analysis of the results
- The tasks will be done under supervision
What are the tasks expected to be accomplished by the student?
Students are expected to participate in experiments including gene cloning, animal cell culture for antibody production, transfection, establishment of stable cell line, purification of recombinant antibody by affinity chromatography, ELISA, SDS-PAGE. Students are also expected to participate in the data analysis of western blotting, co-immunoprecipitation, flow cytometry, fluorescence resonance energy transfer (FRET) assay, confocal microscopy, surface plasmon resonance (SPR) and reporter gene assay.
Will there be any theoretical teaching provided (preliminary readings, lectures, courses, seminars etc)
Informal and personal meetings
What is expected from the student at the end of the research exchange? What will be the general outcome of the student?
- No specific outcome is expected
What skills are required of the student? Is there any special knowledge or a certain level of studies needed?
No specific skills are not required. Subjects passed: Biochemistry and Molecular biology
Are there any legal limitations in the student’s involvement
Type of students accepted
This project accepts:
- Medical students
- Graduated students (less than 6 months)
- Pre-Medical students from the American-British system
- Students in biomedical fields
- Park H; Kim M; Seo Y; Ham Y; Cho MY; Kwon MH. Cytosolic Internalization of Anti-DNA Antibodies by Human Monocytes Induces Production of Pro-inflammatory Cytokines Independently of the Tripartite Motif-Containing 21 (TRIM21)-Mediated Pathway. Front Immunol. 2018 Sep 4;9:2019.
- Park H; Kim M; Kim HJ; Lee Y; Seo Y; Pham CD; Lee J; Byun SJ; Kwon MH. Heparan sulfate proteoglycans (HSPGs) and chondroitin sulfate proteoglycans (CSPGs) function as endocytic receptors for an internalizing anti-nucleic acid antibody. Sci Rep. 2017 Oct 30;7(1):14373.