Enhanced residualizing power of radioiodinated immunoconjugate using a bi-functional linker for radioimmunotherapy
- 주제(키워드) Radioimmunotherapy , Radioimmunoconjugate
- 발행기관 고려대학교 대학원
- 지도교수 박세호
- 지도교수 최태현
- 발행년도 2015
- 학위수여년월 2015. 2
- 학위구분 석사
- 학과 일반대학원 생명과학과
- 세부전공 분자의생명과학
- 원문페이지 73 p
- 실제URI http://www.dcollection.net/handler/korea/000000057706
- 본문언어 영어
- 제출원본 000045827695
초록/요약
Radioimmunotherapy (RIT) is an attractive therapeutic concept, aiming to deliver tumoricial radiation doses to tumors without causing significant radiation toxicity to normal tissues. But a poor retention of radioactivity is a major challenge when these mAbs are radioiodinated by the direct electrophilic approach. To circumvent this problem, (N-(4-isothiocyanatobenzyl)-2-(3-(tributyl stannyl) phenyl) acetamide (FCCS12026) which new bi-functional linker was synthesized. To compare difference of the direct radioiodinated mAbs and indirect radioiodinated mAbs which using bi-functional linker in vitro, the in vitro characterization were assessed by cell binding assay and internalization assay in selected 6 cell lines. The results of in vitro experiment shown that [125I]-FCCS12027-Cetuximab has higher value of specific binding, immunoreactive fraction, total binding to total activity ratios, surface % bound, and internalized % bound than [125I]-Cetuximab. Consequently, indirect labeled Cetuximab using bi-functional linker has higher binding affinity in tumor cell lines and increased immunoreactive fraction than direct labeled Cetuximab. Thus, the labeled mAb using novel linker was anticipated to increase tumor retention in vivo compared with directly labeled mAb
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TABLE OF CONTENTS
LIST OF TABLES ……………………….……………….………I
LIST OF FIGURES …………………………..……….………..III
LIST OF ABBREVIATION …………………..……….……V
ABSTRACT ………………………………………………………VI
1. INTRODUCTION ..........................................................1
2. MATERIALS AND METHODS .................................4
2.1 General …………………………………………………………………...4
2.2 Direct radiolabeling …………………………………..…..……5
2.3 Indirect radiolabeling ………………………………….………..….6
2.4 Protein quantitation analysis …………………………..…….….7
2.5 Cell line ……………………………………………………………….....8
2.6 Culture condition and Cell counting ………….……....8
2.7 Cell binding assay ………………………………………….…9
2.8 Internalization assay ………………………………………..10
2.9 Statistical analysis ……………………………………….….11
3. RESULTS ........................................................................12
3.1 Two type of radioiodinated antibody conjugates …….....12
3.1.1 Direct radiolabeled Cetuximab ……………..….……….....….12
3.1.2 InDirect radiolabeled Cetuximab using novel linker …..12
3.2 Assessment of immunoreactive fraction by cell binding assay….………………………………………………………………………....…16
3.3 In vitro internalization assay ………………………..….……32
4. DISCUSSION ………………………………………...........…48
5. REFERENCES …………………………………………..…….52
6. ABSTRACT IN KOREAN …………………………….……58
7. ACKNOWLEDGEMENT ………………………….……....61
