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United States Department of Agriculture

Agricultural Research Service

Title: Construction of Murine Hybridomas Which Secrete Monoclonal Antibodies Specific for Hypodermin A, a Serine Protease of the Common Cattle Grub, Hypoderma Lineatum

item Temeyer, Kevin
item Pruett Jr, John
item Beetham, Patricia

Submitted to: Southwestern Entomologist
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: September 2, 1996
Publication Date: N/A

Interpretive Summary: Three novel cell lines were constructed which secrete chemically pure antibodies in cell cultures. These antibodies recognize and bind to hypodermin A, an enzyme produced by the common cattle grub. Cattle grubs are parasitic insects of cattle which cause economic damage, including damage to meat and hides. Hypodermin A is a component of a new vaccine for cattle which may help reduce damage by cattle grubs. Antibodies produced by the new cell lines can be used to determine how much hypodermin A is present at various stages of industrial production, to concentrate and purify hypodermin A for vaccine preparation or other uses, or to test for the presence of parasitic cattle grubs in cattle by testing for the presence of hypodermin A. These cell lines and their antibodies are new and useful tools to aid in biochemical studies and control of cattle grubs.

Technical Abstract: Murine hybridomas were constructed that secrete monoclonal antibodies specific for hypodermin A (HyA), a serine protease produced by first instar larvae of the common cattle grub, Hypoderma lineatum (Villers). Monoclonal antibodies produced by the three hybridoma cultures, HL1-A1, HL1-A2 and HL1-A3 were each of subclass IgG1. HL1-A1 antibody bound only the native form of HyA, while HL1-A2 and HL1-A3 antibodies bound both native and SDS-denatured forms of HyA. Binding of either HL1-A1, HL1-A2 or HL1-A3 monoclonal antibodies to HyA hindered subsequent binding of any of the other monoclonal antibodies. HL1-A1 monoclonal antibodies were purified and utilized to develop a sandwich ELISA capable of quantitation of HyA from natural or recombinant sources.

Last Modified: 3/27/2015