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

• Published in: Journal of neuroimmunology Vol. 82; no. 2; pp. 200 - 209
• Main Authors: Graus, Y.F, Verschuuren, J.J, Degenhardt, A, van Breda Vriesman, P.J, De Baets, M.H, Posner, J.B, Burton, D.R, Dalmau, J
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 01-03-1998
• Subjects: 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; HuD; Human antibody; Humans; Immunoglobulin Fab Fragments - immunology; Lung Neoplasms - complications; Lung Neoplasms - immunology; Molecular Sequence Data; Nerve Tissue Proteins - immunology; Neurons - immunology; Paraneoplastic syndrome; Paraneoplastic Syndromes - complications; Paraneoplastic Syndromes - immunology; Phage display; Recombinant Proteins; RNA-Binding Proteins - genetics; RNA-Binding Proteins - immunology; Tumor antigen; Phage display; Paraneoplastic syndrome; HuD; Tumor antigen; Human antibody
• ISSN: 0165-5728; 1872-8421
• DOI: 10.1016/S0165-5728(97)00199-9

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 it 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.