title: A new approach of indirect enzyme-linked immunosorbent assay for determination of D-glutamic acid through in situ conjugation.
abstract: We propose a new approach of an indirect enzyme-linked immunosorbent assay (ELISA) for determination of D-glutamic acid (D-Glu) using a monoclonal antibody against D-glutamic acid (D-Glu-MAb), which recognizes D-Glu-glutaraldehyde (GA) molecule but not D-Glu molecule. Human serum albumin (HSA) was coated on an immunoplate and reacted with D-Glu via GA to produce D-Glu-GA-HSA conjugates in situ in the well to be recognized by D-Glu-MAb, which enabled the development of an indirect ELISA for the determination of free D-Glu. In this indirect ELISA, D-Glu can be specifically detected with limit of detection of 7.81 μ g/mL. Since anti-conjugate antibodies are often produced, even though anti-hapten antibodies are desired, this new approach could be very useful as an application of anti-conjugate antibodies to the development of quantitative analysis for detecting hapten.
authors.affiliation: a Graduate School of Pharmaceutical Sciences , Kyushu University , Higashi-ku , Fukuoka , Japan.
authors.forename: Seiichi
authors.initials: S
authors.lastname: Sakamoto
authors.affiliation: a Graduate School of Pharmaceutical Sciences , Kyushu University , Higashi-ku , Fukuoka , Japan.
authors.forename: Rika
authors.initials: R
authors.lastname: Nagamitsu
authors.affiliation: a Graduate School of Pharmaceutical Sciences , Kyushu University , Higashi-ku , Fukuoka , Japan.
authors.forename: Yurino
authors.initials: Y
authors.lastname: Matsuura
authors.affiliation: a Graduate School of Pharmaceutical Sciences , Kyushu University , Higashi-ku , Fukuoka , Japan.
authors.forename: Yumi
authors.initials: Y
authors.lastname: Tsuneura
authors.affiliation: a Graduate School of Pharmaceutical Sciences , Kyushu University , Higashi-ku , Fukuoka , Japan.
authors.forename: Hitoshi
authors.initials: H
authors.lastname: Kurose
authors.affiliation: a Graduate School of Pharmaceutical Sciences , Kyushu University , Higashi-ku , Fukuoka , Japan.
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Tanaka
authors.affiliation: a Graduate School of Pharmaceutical Sciences , Kyushu University , Higashi-ku , Fukuoka , Japan.
authors.forename: Satoshi
authors.initials: S
authors.lastname: Morimoto
mesh_terms: D000911:Antibodies, Monoclonal / Q000276:immunology*; D000937:Antigen-Antibody Reactions; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D005971:Glutamates / Q000032:analysis* / Q000737:chemistry*; D005976:Glutaral / Q000737:chemistry* / Q000276:immunology*; D006801:Humans; D015394:Molecular Structure; D012709:Serum Albumin / Q000737:chemistry / Q000276:immunology
chemical_list: D000911:Antibodies, Monoclonal; D005971:Glutamates; D012709:Serum Albumin; D005976:Glutaral
keywords: D-glutamic acid (D-Glu); enzyme-linked immunosorbent assay (ELISA); glutaraldehyde (GA); in situ conjugation; monoclonal antibody (MAb)