title: Integrating scFv into xMAP Assays for the Detection of Marine Toxins.
abstract: Marine toxins, such as saxitoxin and domoic acid are associated with algae blooms and can bioaccumulate in shell fish which present both health and economic concerns. The ability to detect the presence of toxin is paramount for the administration of the correct supportive care in case of intoxication; environmental monitoring to detect the presence of toxin is also important for prevention of intoxication. Immunoassays are one tool that has successfully been applied to the detection of marine toxins. Herein, we had the variable regions of two saxitoxin binding monoclonal antibodies sequenced and used the information to produce recombinant constructs that consist of linked heavy and light variable domains that make up the binding domains of the antibodies (scFv). Recombinantly produced binding elements such as scFv provide an alternative to traditional antibodies and serve to "preserve" monoclonal antibodies as they can be easily recreated from their sequence data. In this paper, we combined the anti-saxitoxin scFv developed here with a previously developed anti-domoic acid scFv and demonstrated their utility in a microsphere-based competitive immunoassay format. In addition to detection in buffer, we demonstrated equivalent sensitivity in oyster and scallop matrices. The potential for multiplexed detection using scFvs in this immunoassay format is demonstrated.
authors.affiliation: U.S. Naval Research Laboratory, Center for Biomolecular Science and Engineering, 4555 Overlook Ave SW, Washington, DC 20375, USA. lisa.shriverlake@nrl.navy.mil.
authors.forename: Lisa C
authors.initials: LC
authors.lastname: Shriver-Lake
authors.affiliation: U.S. Naval Research Laboratory, Center for Biomolecular Science and Engineering, 4555 Overlook Ave SW, Washington, DC 20375, USA. jinny.liu@nrl.navy.mil.
authors.forename: Jinny L
authors.initials: JL
authors.lastname: Liu
authors.affiliation: Nova Research Inc., 1900 Elkin St., Suite 230, Alexandria, VA 22308, USA. plee142@su.edu.
authors.forename: P Audrey
authors.initials: PA
authors.lastname: Brozozog Lee
authors.affiliation: U.S. Naval Research Laboratory, Center for Biomolecular Science and Engineering, 4555 Overlook Ave SW, Washington, DC 20375, USA. ellen.goldman@nrl.navy.mil.
authors.forename: Ellen R
authors.initials: ER
authors.lastname: Goldman
authors.affiliation: Chair of Hygiene and Technology of Milk, LMU München, Schönleutnerstraße 8/219, 85764 Oberschleißheim, Germany. r.dietrich@mh.vetmed.uni-muenchen.de.
authors.forename: Richard
authors.initials: R
authors.lastname: Dietrich
authors.affiliation: Chair of Hygiene and Technology of Milk, LMU München, Schönleutnerstraße 8/219, 85764 Oberschleißheim, Germany. e.maertlbauer@mh.vetmed.uni-muenchen.de.
authors.forename: Erwin
authors.initials: E
authors.lastname: Märtlbauer
authors.affiliation: U.S. Naval Research Laboratory, Center for Biomolecular Science and Engineering, 4555 Overlook Ave SW, Washington, DC 20375, USA. george.anderson@nrl.navy.mil.
authors.forename: George P
authors.initials: GP
authors.lastname: Anderson
mesh_terms: D000818:Animals; D005506:Food Contamination / Q000032:analysis; D007118:Immunoassay; D007074:Immunoglobulin G / Q000276:immunology; D007608:Kainic Acid / Q000031:analogs & derivatives* / Q000032:analysis / Q000276:immunology; D010125:Ostreidae; D049897:Pectinidae; D012530:Saxitoxin / Q000032:analysis* / Q000276:immunology; D057127:Single-Chain Antibodies / Q000276:immunology*
chemical_list: D007074:Immunoglobulin G; D057127:Single-Chain Antibodies; D012530:Saxitoxin; C012301:domoic acid; D007608:Kainic Acid
keywords: domoic acid; saxitoxin; single chain antibody; xMAP