Selection of recombinant anti-HuD Fab fragments from a phage display antibody library of a lung cancer patient with paraneoplastic encephalomyelitis

J Neuroimmunol. 1998 Mar 1;82(2):200-9. doi: 10.1016/s0165-5728(97)00199-9.

Abstract

Antibodies against the HuD antigen expressed in small-cell lung cancer (SCLC) cross-react with proteins expressed in neurons of the central and peripheral nervous system and are associated with paraneoplastic encephalomyelitis and sensory neuropathy (PEM/SN). We isolated anti-HuD Fab fragments from an antibody phage display library that was constructed from mRNA of a metastatic lymph node from a patient with SCLC and Pem/SN. Fab GLN495 recognized HuD and other related proteins (HuC and Hel-N1, or Hu antigens) in immunoblots of these recombinant proteins and in immunohistochemical and Western blot analysis of SCLC and neurons. Fab GLN495 inhibited up to 75% of the anti-Hu antibodies of the patient from which it was derived, suggesting that recognizes a dominant epitope in the polyclonal anti-Hu antibody response. Fab GLN495 also competed with anti-Hu sera from most but not all patients with PEM/SN, indicating that the same epitope is recognized by a large subgroup of patients. Human monoclonal anti-HuD antibodies may be useful in diagnosis of HuD expressing tumors and in clarifying the autoimmune etiology of PEM/SN. This study, the first to demonstrate that tumor specific recombinant antibodies can be isolated from metastatic lymph node tissue, shows that this approach may be generally applicable to isolate human antibodies against tumor specific antigens.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Amino Acid Sequence
  • Antibodies / immunology
  • Bacteriophages / immunology
  • Carcinoma, Small Cell / complications
  • Carcinoma, Small Cell / immunology*
  • ELAV Proteins
  • ELAV-Like Protein 4
  • Encephalomyelitis / complications
  • Encephalomyelitis / immunology*
  • Epitopes / immunology
  • Gene Deletion
  • Humans
  • Immunoglobulin Fab Fragments / immunology*
  • Lung Neoplasms / complications
  • Lung Neoplasms / immunology*
  • Molecular Sequence Data
  • Nerve Tissue Proteins / immunology
  • Neurons / immunology
  • Paraneoplastic Syndromes / complications
  • Paraneoplastic Syndromes / immunology*
  • RNA-Binding Proteins / genetics
  • RNA-Binding Proteins / immunology*
  • Recombinant Proteins

Substances

  • Antibodies
  • ELAV Proteins
  • ELAV-Like Protein 4
  • ELAVL4 protein, human
  • Epitopes
  • Immunoglobulin Fab Fragments
  • Nerve Tissue Proteins
  • RNA-Binding Proteins
  • Recombinant Proteins