title: Development of Human Monoclonal Antibodies Against Respiratory Syncytial Virus Using a High Efficiency Human Hybridoma Technique.
abstract: Human monoclonal antibodies against RSV have high potential for use as prophylaxis or therapeutic molecules, and they also can be used to define the structure of protective epitopes for rational vaccine design. In the past, however, isolation of human monoclonal antibodies was difficult and inefficient. Here, we describe contemporary methods for activation and proliferation of primary human memory B cells followed by cytofusion to non-secreting myeloma cells by dielectrophoresis to generate human hybridomas secreting RSV-specific monoclonal antibodies. We also provide experimental methods for screening human B cell lines to obtain RSV-specific lines, especially lines secreting neutralizing antibodies.
authors.affiliation: Department of Pathology, Microbiology and Immunology,, Vanderbilt University Medical Center, Vanderbilt University, Nashville, TN, 37232, USA.|The Vanderbilt Vaccine Center, Vanderbilt University Medical Center, Vanderbilt University, 2213 Garland Avenue, Nashville, TN, 37232-0417, USA.
authors.forename: Gabriela
authors.initials: G
authors.lastname: Alvarado
authors.affiliation: Department of Pathology, Microbiology and Immunology,, Vanderbilt University Medical Center, Vanderbilt University, Nashville, TN, 37232, USA. james.crowe@vanderbilt.edu.|The Vanderbilt Vaccine Center, Vanderbilt University Medical Center, Vanderbilt University, 2213 Garland Avenue, Nashville, TN, 37232-0417, USA. james.crowe@vanderbilt.edu.|Department of Pediatrics, The Vanderbilt Vaccine Center, Vanderbilt University Medical Center, Vanderbilt University, Nashville, TN, 37232, USA. james.crowe@vanderbilt.edu.
authors.forename: James E
authors.initials: JE
authors.lastname: Crowe
mesh_terms: D000911:Antibodies, Monoclonal / Q000378:metabolism*; D057134:Antibodies, Neutralizing / Q000378:metabolism; D001402:B-Lymphocytes / Q000166:cytology* / Q000276:immunology; D002460:Cell Line; D049109:Cell Proliferation; D006801:Humans; D006825:Hybridomas / Q000166:cytology* / Q000276:immunology; D008213:Lymphocyte Activation; D009500:Neutralization Tests; D018113:Respiratory Syncytial Virus, Human / Q000276:immunology*
chemical_list: D000911:Antibodies, Monoclonal; D057134:Antibodies, Neutralizing
keywords: Hybridoma; Monoclonal antibodies; Respiratory syncytial virus; Virus neutralization