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A New Format of Single Chain Tri-specific Antibody with Diminished Molecular Size Efficiently Induces Ovarian Tumor Cell Killing

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Abstract

A combination of bi-specific antibodies (BsAb), anti-tumor×anti-CD3 and anti-tumor×anti-CD28, is effective in vitro and in vivo, whereas production of two kinds of bi-specific antibodies is labor intensive and administration is complicated. Accordingly, we previously developed a new model of single chain tri-specific antibody (scTsAb), sTRI, which linked both the CD3 and CD28 signals for T-cell activation in one molecule, and demonstrated its capacity for triggering T-cells to kill ovary tumor cells. To improve the pharmacokinetics further and decrease the immunogenicity of scTsAb, we have now generated a new format of scTsAb, TR3H, whose molecular size is smaller than sTRI. Here we describe the construction, purification and characterization of TR3H. TR3H scTsAb bound to effector cells and tumor target cells specifically and induced redirected lyses of ovary tumor cells through freshly isolated, unstimulated human peripheral blood lymphocytes (PBLs). This new format of scTsAb possesses properties that support its potential as a new tumor immunotherapeutic agent.

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Abbreviations

EC50 :

half-maximal effective concentration

Fc:

fragment crystallizable

Fv:

variable fragment of antibody

HSA:

human serum albumin

mAb:

monoclonal antibody

OC:

ovarian cancer

PBL:

peripheral blood lymphocyte

PBMC:

peripheral blood mononuclear cells

scBsAb:

single chain bi-specific antibody

ScFv:

single chain Fv

scTsAb:

single chain tri-specific antibody

TAA:

tumor-associated antigen

TBS:

tris-buffered saline

TCR:

T-cell receptor

VH:

variable region of heavy chain

VL:

variable region of light chain

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Authors and Affiliations

  1. Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Datun Road, 100101, Beijing, China

    Jing Liu, Qi Zhao, Xiangbin Wang, Liping Song, Zhang Zhong, Qing Lin & Hualiang Huang

  2. College of Life Sciences, Lanzhou University, 730000, Lanzhou, China

    Baofeng Zhao

  3. Pharmaceutical Department, Shenyang Pharmaceutical University, 110015, Shenyang, China

    Julong Cheng

Authors
  1. Jing Liu
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  2. Qi Zhao
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  3. Baofeng Zhao
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  4. Julong Cheng
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  5. Xiangbin Wang
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  6. Liping Song
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  7. Zhang Zhong
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  8. Qing Lin
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  9. Hualiang Huang
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Corresponding author

Correspondence to Hualiang Huang.

Additional information

Revisions requested August 2005; Revisions received 14 September 2005

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Liu, J., Zhao, Q., Zhao, B. et al. A New Format of Single Chain Tri-specific Antibody with Diminished Molecular Size Efficiently Induces Ovarian Tumor Cell Killing. Biotechnol Lett 27, 1821–1827 (2005). https://doi.org/10.1007/s10529-005-6732-4

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  • DOI: https://doi.org/10.1007/s10529-005-6732-4

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