Title

Ligand-receptor interaction quantified by newly engineered fluorescence proteins

Alternative Title

새로이 설계된 형광 단백질을 활용한 리간드-수용체 상호작용의 정량화

DGIST Authors

Eunho Kang ; Chang-Hun Lee ; Jin Hae Kim

Advisor

이창훈

Co-Advisor(s)

Jin Hae Kim

Issued Date

2025

Awarded Date

2025-02-01

Citation

Eunho Kang. (2025). Ligand-receptor interaction quantified by newly engineered fluorescence proteins. doi: 10.22677/THESIS.200000844467

Type

Thesis

Description

BiFC, Erythropoietin receptor, Protein engineering, Protein modelling

Table Of Contents

I. Introduction 1

II. Materials and methods 4
1. Designing biosensor system with spGFP assembly system 4
2. Transformation, culture and overexpression on e.coli system 4
3. Purification of target protein 5
4. Normalizing molar ration of sensor molecules 6
5. Detection of signal emission from biosensor system. 7
6. Cloning of target gene 7
7. Design of novel peptide ligand of erythropoietin receptor 8
8. Statistical analysis 9

III. Results 10
1. Biosensor property configuration 10
2. Biosensor activity confirmation 12
3. Evaluating activity of denatured EPO 15
4. Testing EPO’s interaction between spGFP domains 17
5. Cloning another biosensor with different receptor system 19
6. Ligand design with RFdiffusion – single binding peptides 21
7. Dimerization screening of designed ligands by BiFC – single binding peptides 23
8. Ligand design with RFdiffusion – dual binding peptides 25
9. Dimerization screening of designed ligands by BiFC – dual binding peptides 26
IV. Discussion 30
V. Conclusion 33
VI. References 35

URI

http://hdl.handle.net/20.500.11750/58013
http://dgist.dcollection.net/common/orgView/200000844467

DOI

10.22677/THESIS.200000844467

Degree

Master

Department

Department of New Biology

Publisher

DGIST

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