본문 바로가기
KERD
논문KCI 등재

Isolation and Characterization of Human scFv Molecules Specific for Recombinant Human Heat Shock Protein (HSP) 70.1

이재선|백현정|서정선|차상훈
대한면역학회 (The Korean Association of Immunologists)|2004.03|Immune Network|제4권 제1호|pp. 7-15

2

조회

0

다운로드

0

인용

0

참고문헌

초록

Abstract

Background: The heat shock proteins (HSPs) play an important role in cellular protection mechanisms against physical or chemical stresses. In this study scFv antibodies specific for human HSP70.1 were isolated from a semi-synthetic human scFv library with the ultimate goal of developing anti-HSP70.1 intracellular antibody (intrabody) that may offer an attractive alternative to gene targeting to study the function of the protein in cells. Methods: A semi-synthetic human scFv display library (5 108 size) was constructed using pCANTAB-5E vector and the selection of the library against bacterially expressed recombinant human HSP70.1 was attempted by panning. Results: Three positive clones specific for recombinant HSP70.1 were identified. All three clones used VH subgroup III. On the other hand, VL of two clones belonged to the kappa light chain subgroup I, but the other utilized Vk subgroup IV Interestingly, these scFv molecules specifically reacted to the recombinant HSP70.1, yet failed to recognize native HSP70 induced in U937 human monocytic cells by heat treatment. Conclusion: Our results indicated that affinity selection of an scFv phage display library using recombinant antigens produced in E. coli might not guarantee the isolation of scFv antibody molecules specific for a native form of the antigen. Therefore, the source of target antigens needs to be chosen carefully in order to isolate biofunctional antibody molecules. (Immune Network 2004;4(1):7-15)

참고문헌 (33)

  1. "Human immune response to monoclonal antibodies", 15, pp.42~52 (1994)
  2. "Antibody humanization using monovalent phage display", 272, pp.10678~10684 (1997)
  3. "Selection and evolution of high- affinity human anti-viral antibodies", 14, pp.230~234 (1996)
  4. "By-passing immunization:Human antibodies from V-gene libraries displayed on phage", 222, pp.581~597 (1991)
  5. "By-passing immunization:Human antibodies from synthetic repertoires of germline VH gene segments rearranged in vitro", 227, pp.381~388 (1992)
  6. "Intracellular antibodies:development and therapeutic potential", 13, pp.306~310 (1995)
  7. "Phenotypic knockout of the high-affinity human interleukin 2 receptor by intracellular single-chain antibodies against the alpha subunit of the receptor", 92, pp.3137~3141 (1995)
  8. "Intracellular expression of single chain antibodies reverts ErbB-2 transformation", 269, pp.23931~23936 (1994)
  9. "A cancer gene therapy approach utilizing an anti-erbB-2 single-chain antibody-encoding adenovirus", 6, pp.3081~3087 (2000)
  10. "Intracellular expression of anti-p21ras single chain Fv fragments inhibits meiotic maturation of xenopus oocytes", 197, pp.422~427 (1993)
  11. "Combined intra- and extracellular immunization against human immunodeficiency virus type 1 infection with a human anti-gp120 antibody", 91, pp.5932~5936 (1994)
  12. "Inhibition of human immunodeficiency virus type 1 replication in vitro by a novel combination of anti-Tat single-chain intrabodies and NF-kappa B antagonists", 71, pp.6486~6494 (1997)
  13. "Human anti- HIV-1 tat sFv intrabodies for gene therapy of advanced HIV- 1-infection and AIDS", 231, pp.223~238 (1999)
  14. "Redox state of single chain Fv fragments targeted to the endoplasmic reticulum, cytosol and mitochondria", 13, pp.1110~1115 (1995)
  15. "Heat shock proteins and the inflammatory response", 856, pp.206~213 (1993)
  16. "Heat shock proteins increase resistance to apoptosis", 223, pp.63~70 (1996)
  17. "Heat shock protein 70 kDa: Molecular biology, biochemistry, and physiology", 80, pp.183~201 (1998)
  18. "Role of the major heat shock proteins as molecular chaperones", 9, pp.601~634 (1993)
  19. "Chromosomal localization of five murine HSP70 gene family members", ge(mics 16), pp.~1 2 3
  20. "Expression of heat shock proteins by isolated mouse spermatogenic cells", 8, pp.3260~3266 (1988)
  21. "Identification and se-quence analysis of a new member of the mouse HSP70 gene family and characterization of its unique cellular and developmental pattern of expression in the male germ line", 8, pp.2925~2932 (1988)
  22. "Impaired repair ability of hsp70 1 KO mouse after UVB irradiation", 28, pp.144~151 (2002)
  23. "Targeted hsp70.1 disruption increases infarction volume after focal cerebral ischemia in mice.", 32, pp.2905~2912 (2001)
  24. "Insights into regulation and function of the major stress-induced hsp70 molecular chaperone in vivo analysis of mice with targeted gene disruption of the hsp70. 1 or hsp70.3 gene", 21, pp.8575~8591 (2001)
  25. "Differential induction of Hsp70-encoding genes in human hematopoietic cells", 276, pp.31713~31719 (2001)
  26. "Construction and screening of antibody display libraries", pp.79~111 (1996), Academic Press
  27. "Phage antibodies:filamentous phage displaying antibody variable domains", 348, pp.552~554 (1990)
  28. "Heat shock protein 70 inhibits apoptosis downstream of cytochrome c release and upstream of caspase-3 activation", 275, pp.25665~25671 (2000)
  29. "Isolation of anti- mesothelin antibodies from a phage display library", 34, pp.9~20 (1997)
  30. "recombinant, fully human monoclonal antibody with antitumor activity constructed from phage-displayed antibody fragments", 17, pp.276~281 (1999)
  31. "Isolation of tumor cell-specific single-chain Fv from immunized mice using phage-antibody libraries and the re-construction of whole antibodies from these antibody fragments", 24, pp.952~958 (1994)
  32. "High-throughput generation and engineering of recombinant human antibodies", pp.67~84 (2001)
  33. "Properties of a panel of single chain variable fragments against potato leafroll virus obtained from two phage display libraries", 81, pp.159~168 (1999)

관련 논문AI추천

Single-Stranded Variable Fragment Gene Libraries Built for Phage Display: An Updated Review of Design, Selection and ApplicationCordeiro Isabelle Bezerra, da Silva Eliza Raquel Duarte, de Alcântara Kelson Ávila Graça, de Andrade Edmar Vaz, de Lima Guerra Corado André, de Souza Caio Coutinho, Glória Juliane Corrêa, Mariúba Luis André Morais한국미생물·생명공학회 · Journal of Microbiology and Biotechnology2025 · 제35권 Expression and Characterization of a Single-Chain Variable Fragment against Human LOX-1 in Escherichia coli and Brevibacillus choshinensisHongyu Xiang, Jun-Yan Xiang, Ling Liu, Ping Kong, Qiuhong Xie, Wei Hu한국미생물·생명공학회 · Journal of Microbiology and Biotechnology2017 · 제27권 제5호Porphyromonas gingivalis의 열충격단백-특이성 단클론항체의 개발김성조, 이니나, 이주연, 최점일대한치주과학회 · Journal of Periodontal & Implant Science2007 · 제37권 제1호Affinity Maturation of an Epidermal Growth Factor Receptor Targeting Human Monoclonal Antibody ER414 by CDR Mutation김세호, Gwang-Won Hong, Ki-Hwan Chang, Min-Soo Kim, Mi-Sun Seo, Yong-Nam Shin대한면역학회 · Immune Network2012 · 제12권 제4호Development of Bispecific Antibody for Cancer Immunotherapy: Focus on T Cell Engaging Antibody김대희, 문다인, 박윤지, 이승우, Tae Nara대한면역학회 · Immune Network2022 · 제22권 제1호Comprehensive Identification of Tumor-associated Antigens via Isolation of Human Monoclonal Antibodies that may be TherapeuticYoshikazu Kurosawa대한면역학회 · Immune Network2009 · 제9권 제1호

이 간행물의 다른 논문 (8건)

이식면역학의 역사적 고찰박정규pp. 1-6Phage Display 기법을 이용한 B형 간염 바이러스 Polymerase의 RNase H 활성을 억제하는 인간 단세포군 항체의 개발정영주, 이성락, 박세광, 최인학, 이영이, 송은경, 김익중pp. 16-22인체 SIP 단백질에 특이적인 단일클론 항체의 특성김진숙, 윤선영, 이영희, 김창남, 김주헌, 주종혁, 권두한, 최인성, 강호범, 김재화pp. 23-30HHD Mice를 이용한 대장암세포유래 펩타이드 특이적 CD8+ T 세포의 입양전이안인숙, 정헌순, 도형기, Lea Eisenbach, Ezra Vadai, Esther Tzehoval, Boaz Tirosh, Francois A. Lemonnier, 도명술pp. 31-37골수성백혈병에서 배양한 수지상세포(Dendritic Cell)에 대한 종양항원 감작법으로 IL-12 첨가와 융합법의 효과김기원, 박석영, 홍영선pp. 38-43
원문 보기