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Targeting acute myeloid cell surface using a recombinant antibody isolated from whole-cell biopanning of a phage display human scFv antibody library

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Abstract

To discover new therapeutic antibodies for treatment of acute myeloid leukemia (AML) without the requirement of a known antigen, a human single-chain variable fragment (scFv) library was used to isolate novel antibody fragments recognizing HL-60 AML cells. After three rounds of biopanning, scFv-expressing phages were selected to test for binding to the target cell by flow cytometry. The clone with highest binding specificity to HL-60 cells (designated y1HL63D6) was further investigated. Fluorescent staining indicated that y1HL63D6 scFv bound to a target located on the cell surface. Whole immunoglobulin, IgG-y1HL63D6 was then generated and tested for the binding against bone marrow mononuclear cells (BMMCs) from AML patients. Significantly higher fluorescent signals were observed for some patient samples when compared to normal BMMCs or non-AML patients’ BMMCs. Next, the IgG-y1HL63D6 format was tested for antibody-dependent cell cytotoxicity (ADCC). The results demonstrated that IgG-y1HL63D6 but not the control antibody, trastuzumab, could mediate specific killing of HL-60 target cells. In conclusion, our results indicate that specific antibodies can be isolated by biopanning whole cells with a non-immunized human scFv antibody phage display library and that the isolated antibody against HL-60 cells showed therapeutic potential.

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Data availability

The datasets generated during and/or analyzed during the current study are available from the corresponding author on reasonable request.

Abbreviations

AML:

Acute myeloid leukemia

scFv:

Single-chain variable fragment

IgG:

Immunoglobulin G

VH:

Variable heavy

VL:

Variable light

HC:

Heavy chain

LC:

Light chain

BMMCs:

Bone marrow mononuclear cells

PBMCs:

Peripheral blood mononuclear cells

ADCC:

Antibody-dependent cell-mediated cytotoxicity

MFI:

Median fluorescent intensity

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Acknowledgements

The authors would like to thank Miss Yuwadee Kitprasong, medical technologist at Sanphasitthiprasong hospital, for patient sample collection. We would also like to thank to all participants who volunteered for this study. We are in dept. of Prof. Dr. Pa-thai Yenchitsomanus from Siriraj Center of Research Excellence for Cancer Immunotherapy, Mahidol University, Thailand for several cell lines and grateful to MY lab members and KAC lab members for excellent technical assistances and advice.

Funding

This research was supported by Thailand Science Research and Innovation (TSRI) (TRF Senior Research grant number RTA6180012) and BIOTEC, the National Science and Technology Development Agency (NSTDA) (grant number P-18–50127), and Ministry of Higher Education, Science, Research and Innovation (MHESI) (grant number 256101A3040017). TS was co-supported by the Royal Golden Jubilee PhD program of Thailand (rgj.trf.or.th) and Suranaree University of Technology (www.sut.ac.th) [grant number PHD/0098/2553]. She also was supported by Newton Fund from British Council, as well as grants from MY Lab. MY was also supported by the Distinguished Research Professor Grant (NRCT 808/2563) of the National Research Council of Thailand.

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Authors and Affiliations

  1. Molecular Biotechnology Laboratory, School of Biotechnology, Institute of Agricultural Technology, Suranaree University of Technology, 111 University Avenue, Nakhon Ratchasima, 30000, Thailand

    Thitima Sumphanapai & Montarop Yamabhai

  2. University College London Cancer Institute, 72 Huntley Street, London, WC1E 6BT, UK

    Kerry Chester & Jenny Yeung

  3. Hematology Unit, Sunpasitthiprasong Hospital, Ubon Ratchathani, 34000, Thailand

    Surasak Sawatnatee

  4. University College London Great Ormond Street Institute of Child Health, 30 Guilford Street, London, WC1N 1EH, UK

    Jenny Yeung

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  1. Thitima Sumphanapai
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Contributions

Thitima Sumphanapai performed the experiments, data collection, and writing of the initial draft. Surasak Sawatnatee provided patient samples and had a consultant/advisory role. Jenny Yeung conceived the experiments, supervised assay techniques, and revised the manuscript. Kerry Chester conceived the experiments and provided the critical review, commentary, and revision. Montarop Yamabhai conceived the experiments, conceptualization, supervision, edited the manuscript, and acquisition of the financial support for the project leading to this publication. All authors reviewed the manuscript.

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Correspondence to Montarop Yamabhai.

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The authors declare that they have no competing interests.

Ethics approval

The patient samples were collected under the approval of the Ethical Review Board of Sanphasitthiprasong hospital, Ubon Ratchathani, Thailand. The peripheral blood samples of healthy donors were obtained under the approval of the research ethics board of the Suranaree University of Technology, Nakhon Ratchasima, Thailand, in accordance with the Declaration of Helsinki.

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All samples analyzed in this study were obtained after informed consent of the donors.

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Sumphanapai, T., Chester, K., Sawatnatee, S. et al. Targeting acute myeloid cell surface using a recombinant antibody isolated from whole-cell biopanning of a phage display human scFv antibody library. Med Oncol 39, 205 (2022). https://doi.org/10.1007/s12032-022-01806-9

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