Generation of Human Antibodies Against a Specific Peptide: a Novel Strategy Based on Human Cells

Anticancer Res. 2016 Sep;36(9):4613-21. doi: 10.21873/anticanres.11011.

Abstract

Background/aim: We developed a novel platform to produce fully human antibodies that could be used as tools in cancer research. This study was based on maturation of B-cells into plasma cells and the subsequent secretion of fully human antibodies against a 9-mer peptide.

Materials and methods: A peptide was introduced to dendritic cells, followed by co-cultivation with cluster of differentiation 4 (CD4+) cells from healthy donors and CD19(+) cells from patients with cancer. In order to detect plasma cells, we used microscopy, flow cytometry, enzyme-linked immunosorbent assays (ELISAs), and western blotting.

Results: Oval-shaped cells with eccentric nuclei were found which expressed high levels of CD27 (up to 73%) and CD138 (up to 31%). An antibody against the selected peptide was detected in the culture supernatants of all samples according to ELISA and western blot analyses.

Conclusion: This platform successfully generated fully human antibodies against a specific epitope.

Keywords: Monoclonal antibodies; breast cancer.

MeSH terms

  • Adult
  • Aged
  • Aged, 80 and over
  • Antibodies, Monoclonal / chemistry*
  • Antibodies, Monoclonal / immunology
  • Antigens, CD19 / metabolism
  • Breast Neoplasms / immunology*
  • Breast Neoplasms / therapy*
  • CD4-Positive T-Lymphocytes / cytology
  • Coculture Techniques
  • Dendritic Cells / cytology
  • Enzyme-Linked Immunosorbent Assay
  • Epitopes / chemistry
  • Female
  • Flow Cytometry
  • Humans
  • Leukocytes, Mononuclear / cytology
  • Male
  • Middle Aged
  • Peptides / immunology*
  • Syndecan-1 / metabolism
  • Tumor Necrosis Factor Receptor Superfamily, Member 7 / metabolism
  • Young Adult

Substances

  • Antibodies, Monoclonal
  • Antigens, CD19
  • CD19 molecule, human
  • Epitopes
  • Peptides
  • SDC1 protein, human
  • Syndecan-1
  • Tumor Necrosis Factor Receptor Superfamily, Member 7