title: Production of functional antibodies generated in a nonlytic insect cell expression system.
abstract: A monoclonal antibody directed against the type 2 adenovirus (Ad2) penton base protein was cloned and expressed in Spodoptera frugiperda (Sf9) cells using a nonlytic vector system. The coding sequences for the immunoglobulin light and heavy chains were placed under the control of the Orgyia pseudotsugata multicapsid nucleopolyhedrosis virus immediate-early 2 (OpIE2) promoter. Transfected Sf9 cells continuously secreted the antibody which retained the ability to recognize both native and recombinant Ad2 penton base proteins. Bifunctional penton base antibodies were also generated by fusing a gene for a growth factor or a cytokine at the 3' end of the Ig constant heavy chain domain. The quantity and activity of recombinant antibodies generated in the nonlytic insect cell system could be determined relatively quickly compared to other expression systems. Moreover, these recombinant proteins were not subjected to proteolytic degradation as frequently occurs during baculovirus-mediated cell lysis and the levels of recombinant antibodies produced in the nonlytic system were comparable to those reported for cytolytic baculovirus vectors.
authors.affiliation: Department of Immunology, The Scripps Research Institute, 10550 North Torrey Pines Road, La Jolla, CA 92037, USA.
authors.forename: E
authors.initials: E
authors.lastname: Li
authors.forename: S L
authors.initials: SL
authors.lastname: Brown
authors.forename: C S
authors.initials: CS
authors.lastname: Dolman
authors.forename: G B
authors.initials: GB
authors.lastname: Brown
authors.forename: G R
authors.initials: GR
authors.lastname: Nemerow
mesh_terms: D000256:Adenoviridae / Q000276:immunology; D000595:Amino Acid Sequence; D000818:Animals; D002213:Capsid / Q000276:immunology; D036022:Capsid Proteins; D002460:Cell Line; D003001:Cloning, Molecular / Q000379:methods; D018076:DNA, Complementary; D004591:Electrophoresis, Polyacrylamide Gel; D005822:Genetic Vectors; D006825:Hybridomas; D007074:Immunoglobulin G / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification; D007143:Immunoglobulin Heavy Chains / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification; D007147:Immunoglobulin Light Chains / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification; D051379:Mice; D008969:Molecular Sequence Data; D009036:Moths; D017924:Nucleopolyhedroviruses / Q000235:genetics; D011401:Promoter Regions, Genetic; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000302:isolation & purification; D018411:Spodoptera; D014162:Transfection / Q000379:methods
chemical_list: D036022:Capsid Proteins; D018076:DNA, Complementary; D007074:Immunoglobulin G; D007143:Immunoglobulin Heavy Chains; D007147:Immunoglobulin Light Chains; D011994:Recombinant Proteins; C043322:penton protein, adenovirus