A new approach of indirect enzyme-linked immunosorbent assay for determination of D-glutamic acid through in situ conjugation

Seiichi Sakamoto, Rika Nagamitsu, Yurino Matsuura, Yumi Tsuneura, Hitoshi Kurose, Hiroyuki Tanaka, Satoshi Morimoto

Research output: Contribution to journalArticle

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 μg/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.

Original languageEnglish
Pages (from-to)296-306
Number of pages11
JournalJournal of Immunoassay and Immunochemistry
Volume37
Issue number3
DOIs
Publication statusPublished - May 3 2016

All Science Journal Classification (ASJC) codes

  • Immunology and Allergy
  • Immunology
  • Clinical Biochemistry
  • Medical Laboratory Technology

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