title: Antigen itself antibody: an alternative one-step immunoassay for measuring the anti-idiotypic antibody titer.
abstract: Immunoassay designs rely on the great specificity of antibodies and a suitable marker that facilitates generation of a quantitative signal. Currently, there is no reliable method for measuring the titers of an anti-idiotypic antibody. Our initial attempt to measure titers of mouse anti-idiotypic antibody after idiotypic vaccination with HM-1 killer toxin neutralizing monoclonal antibody (nmAb-KT) failed. Because the injected antigen, nmAb-KT, is a mouse IgG, using a commercial antibody to measure the antibody titer always gave a false positive signal against control mouse serum antibody in parallel with the antigen-treated immunized serum antibodies. To get a reliable and clearly differentiable signal by ELISA, idiotypic antigen was labeled with HRP and HRP-conjugated-nmAb-KT used to measure the antibody titers in the antigen-treated mice. Compared with control mice, signals were found in high anti-nmAb-KT IgG responses in test mice; however, untreated control mice had a significant amount of purified non-specific IgG. This method is amenable to long read lengths and will likely enable anti-idiotypic antibody titer measurement in a more specific and cost effective way without requiring commercial antibody.
authors.affiliation: Department of Biochemistry, Faculty of Pharmaceutical Sciences, Niigata University of Pharmacy and Applied Life Sciences, 265-1 Higashijima, Niigata 956-8603; Department of Pathology, School of Medicine, Case Western Reserve University, Cleveland, OH 44106.
authors.forename: M Enamul
authors.initials: ME
authors.lastname: Kabir
authors.forename: Senthilkumar
authors.initials: S
authors.lastname: Krishnaswamy
authors.forename: Dakshnamurthy
authors.initials: D
authors.lastname: Selvakumar
authors.forename: Masahiko
authors.initials: M
authors.lastname: Miyamoto
authors.forename: Yasuhiro
authors.initials: Y
authors.lastname: Furuichi
authors.forename: Tadazumi
authors.initials: T
authors.lastname: Komiyama
mesh_terms: D000818:Animals; D000888:Antibodies, Anti-Idiotypic / Q000097:blood*; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D005260:Female; D008807:Mice, Inbred BALB C
chemical_list: D000888:Antibodies, Anti-Idiotypic
keywords: anti-idiotypic antibody; killer toxin neutralizing monoclonal antibody; one-step immunoassay; titer measurement