Development of an artificial antibody specific for HLA/peptide complex derived from cancer stem-like cell/cancer-initiating cell antigen DNAJB8

• Published in: British journal of cancer 2020-10, Vol.123 (9), p.1387-1394
• Main Authors: Tadano, Hiroki, Tsukahara, Tomohide, Mizushima, Emi, Akamatsu, Asuka, Watanabe, Kazue, Nojima, Iyori, Kubo, Terufumi, Kanaseki, Takayuki, Hirohashi, Yoshihiko, Sato, Noriyuki, Torigoe, Toshihiko
• Format: Article
• Language: English
• Subjects: 631/250/580/1884/2323; 692/4017; Affinity; Antibodies; Antibody Specificity; Antibody-Dependent Cell Cytotoxicity; Antigen-presenting cells; Antigens; Antigens, Neoplasm - genetics; Antigens, Neoplasm - metabolism; Biomedical and Life Sciences; Biomedicine; Bispecific antibodies; Bone Neoplasms - immunology; Bone Neoplasms - pathology; Bone Neoplasms - therapy; Cancer Research; Cancer Vaccines - biosynthesis; Cancer Vaccines - therapeutic use; Carcinoma, Renal Cell - immunology; Carcinoma, Renal Cell - pathology; Carcinoma, Renal Cell - therapy; Cell Line, Tumor; Cell Transformation, Neoplastic - genetics; Cell Transformation, Neoplastic - metabolism; Cytotoxicity; Drug Resistance; Epidemiology; Flow cytometry; HEK293 Cells; Histocompatibility antigen HLA; HLA-A24 Antigen - genetics; HLA-A24 Antigen - immunology; HLA-A24 Antigen - metabolism; HSP40 Heat-Shock Proteins - genetics; HSP40 Heat-Shock Proteins - immunology; HSP40 Heat-Shock Proteins - metabolism; HT29 Cells; Humans; Immune response; Immunoglobulin G; Immunotherapy; Immunotherapy - methods; Kidney cancer; Kidney Neoplasms - immunology; Kidney Neoplasms - pathology; Kidney Neoplasms - therapy; Lymphocytes T; Molecular Chaperones - genetics; Molecular Chaperones - immunology; Molecular Chaperones - metabolism; Molecular Medicine; Neoplastic Stem Cells - immunology; Neoplastic Stem Cells - metabolism; Neoplastic Stem Cells - pathology; Nerve Tissue Proteins - genetics; Nerve Tissue Proteins - immunology; Nerve Tissue Proteins - metabolism; Oncology; Osteosarcoma; Osteosarcoma - immunology; Osteosarcoma - pathology; Osteosarcoma - therapy; Panning; Peptide Fragments - immunology; Peptide Fragments - metabolism; Peptide Library; Peptides; Phages; Protein Engineering - methods; Renal cell carcinoma; Single-Chain Antibodies - biosynthesis; Single-Chain Antibodies - therapeutic use; Surface plasmon resonance; Tumor cell lines; Tumors; Vaccination
• ISSN: 0007-0920; 1532-1827
• DOI: 10.1038/s41416-020-1017-1

Background Peptide-vaccination therapy targeting tumour-associated antigens can elicit immune responses, but cannot be used to eliminate large tumour burden. In this study, we developed a therapeutic single-chain variable-fragment (scFv) antibody that recognises the cancer stem-like cell/cancer-initiating cell (CSC/CIC) antigen, DNAJB8. Methods We screened scFv clones reacting with HLA-A24:20/DNAJB8-derived peptide (DNAJB8_143) complex using naive scFv phage-display libraries. Reactivity and affinity of scFv clones against the cognate antigen were quantified using FACS and surface plasmon resonance. Candidate scFv clones were engineered to human IgG1 (hIgG1) and T-cell-engaging bispecific antibody (CD3xJB8). Complement-dependent cytotoxicity (CDC) and bispecific antibody-dependent cellular cytotoxicity (BADCC) were assessed. Results scFv clones A10 and B10 were isolated after bio-panning. Both A10-hIgG1 and B10-hIgG1 reacted with DNAJB8-143 peptide-pulsed antigen-presenting cells and HLA-A24(+)/DNAJB8(+) renal cell carcinoma and osteosarcoma cell lines. A10-hIgG1 and B10-hIgG1 showed strong affinity with the cognate HLA/peptide complex ( K D  = 2.96 × 10 −9  M and 5.04 × 10 −9  M, respectively). A10-hIgG1 and B10-hIgG1 showed CDC against HLA-A24(+)/DNAJB8(+) cell lines. B10-(CD3xJB8) showed superior BADCC to A10-(CD3xJB8). Conclusion We isolated artificial scFv antibodies reactive to CSC/CIC antigen DNAJB8-derived peptide naturally present on renal cell carcinoma and sarcoma. Immunotherapy using these engineered antibodies could be promising.