title: High efficiency creation of human monoclonal antibody-producing hybridomas.
abstract: The native human antibody repertoire holds unexplored potential for the development of novel monoclonal antibody therapeutics. Current techniques that fuse immortal cells and primary B-lymphocytes are sub-optimal for the routine production of hybridomas that secrete human monoclonal antibodies. We have found that a murine cell line that ectopically expresses murine interleukin-6 (mIL-6) and human telomerase (hTERT) efficiently forms stable human antibody-secreting heterohybridomas through cell fusion with primary human B-lymphocytes. The hybrid cells maintain secretion of human antibodies derived from the primary B-lymphocytes through multiple rounds of cloning. Using splenic B-lymphocytes from a patient immunized with a Streptococcus pneumoniae capsular polysaccharide vaccine, we have succeeded in creating hybridomas that secrete human monoclonal antibodies specific for S. pneumoniae antigens. Using peripheral blood lymphocytes, we have similarly cloned a human antibody that binds a viral antigen. These experiments establish that SP2/0-derived cell lines ectopically expressing mIL-6 and hTERT will enable the rapid cloning of native human monoclonal antibodies.
authors.affiliation: The Whitehead Institute for Biomedical Research, 9 Cambridge Center, Cambridge, MA 02142, USA. Scott.Dessain@jefferson.edu
authors.forename: Scott K
authors.initials: SK
authors.lastname: Dessain
authors.forename: Sharad P
authors.initials: SP
authors.lastname: Adekar
authors.forename: Jennifer B
authors.initials: JB
authors.lastname: Stevens
authors.forename: Katherine A
authors.initials: KA
authors.lastname: Carpenter
authors.forename: Maria L
authors.initials: ML
authors.lastname: Skorski
authors.forename: Barry L
authors.initials: BL
authors.lastname: Barnoski
authors.forename: Richard A
authors.initials: RA
authors.lastname: Goldsby
authors.forename: Robert A
authors.initials: RA
authors.lastname: Weinberg
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis* / Q000276:immunology; D000918:Antibody Specificity; D000941:Antigens / Q000276:immunology; D001402:B-Lymphocytes / Q000166:cytology / Q000276:immunology / Q000378:metabolism; D002460:Cell Line; D002877:Chromosomes, Human; D002999:Clone Cells / Q000166:cytology / Q000276:immunology / Q000378:metabolism; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D006825:Hybridomas / Q000166:cytology / Q000276:immunology* / Q000378:metabolism*; D007136:Immunoglobulins / Q000276:immunology / Q000378:metabolism; D015850:Interleukin-6 / Q000378:metabolism; D007621:Karyotyping; D051379:Mice; D008875:Middle Aged; D011135:Polysaccharides, Bacterial / Q000276:immunology; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D013296:Streptococcus pneumoniae / Q000276:immunology; D019098:Telomerase / Q000235:genetics / Q000378:metabolism
chemical_list: D000911:Antibodies, Monoclonal; D000941:Antigens; D007136:Immunoglobulins; D015850:Interleukin-6; D011135:Polysaccharides, Bacterial; D012333:RNA, Messenger; D019098:Telomerase