title: Polyclonal antibody to soman-tyrosine.
abstract: Soman forms a stable, covalent bond with tyrosine 411 of human albumin, with tyrosines 257 and 593 in human transferrin, and with tyrosine in many other proteins. The pinacolyl group of soman is retained, suggesting that pinacolyl methylphosphonate bound to tyrosine could generate specific antibodies. Tyrosine in the pentapeptide RYGRK was covalently modified with soman simply by adding soman to the peptide. The phosphonylated-peptide was linked to keyhole limpet hemocyanin, and the conjugate was injected into rabbits. The polyclonal antiserum recognized soman-labeled human albumin, soman-mouse albumin, and soman human transferrin but not nonphosphonylated control proteins. The soman-labeled tyrosines in these proteins are surrounded by different amino acid sequences, suggesting that the polyclonal recognizes soman-tyrosine independent of the amino acid sequence. Antiserum obtained after 4 antigen injections over a period of 18 weeks was tested in a competition ELISA where it had an IC50 of 10(-11) M. The limit of detection on Western blots was 0.01 μg (15 picomoles) of soman-labeled albumin. In conclusion, a high-affinity, polyclonal antibody that specifically recognizes soman adducts on tyrosine in a variety of proteins has been produced. Such an antibody could be useful for identifying secondary targets of soman toxicity.
authors.affiliation: Eppley Institute, University of Nebraska Medical Center , Omaha, Nebraska 68198-5950, United States.
authors.forename: Bin
authors.initials: B
authors.lastname: Li
authors.forename: Ellen G
authors.initials: EG
authors.lastname: Duysen
authors.forename: Marie-Thérèse
authors.initials: MT
authors.lastname: Froment
authors.forename: Patrick
authors.initials: P
authors.lastname: Masson
authors.forename: Florian
authors.initials: F
authors.lastname: Nachon
authors.forename: Wei
authors.initials: W
authors.lastname: Jiang
authors.forename: Lawrence M
authors.initials: LM
authors.lastname: Schopfer
authors.forename: Geoffrey M
authors.initials: GM
authors.lastname: Thiele
authors.forename: Lynell W
authors.initials: LW
authors.lastname: Klassen
authors.forename: John
authors.initials: J
authors.lastname: Cashman
authors.forename: Gareth R
authors.initials: GR
authors.lastname: Williams
authors.forename: Oksana
authors.initials: O
authors.lastname: Lockridge
mesh_terms: D000818:Animals; D000906:Antibodies / Q000276:immunology*; D000941:Antigens / Q000737:chemistry / Q000276:immunology* / Q000378:metabolism; D004797:Enzyme-Linked Immunosorbent Assay; D006433:Hemocyanins / Q000737:chemistry / Q000276:immunology; D006801:Humans; D051379:Mice; D009842:Oligopeptides / Q000737:chemistry / Q000276:immunology; D011817:Rabbits; D012999:Soman / Q000737:chemistry / Q000276:immunology* / Q000378:metabolism; D014443:Tyrosine / Q000737:chemistry / Q000276:immunology* / Q000378:metabolism
chemical_list: D000906:Antibodies; D000941:Antigens; D009842:Oligopeptides; D014443:Tyrosine; D006433:Hemocyanins; D012999:Soman; C032808:keyhole-limpet hemocyanin