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

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Immunosorbents
Glutamic Acid
Anti-Idiotypic Antibodies
Assays
Haptens
Enzyme-Linked Immunosorbent Assay
Serum Albumin
Antibodies
Enzymes
Molecules
Glutaral
Limit of Detection
Monoclonal Antibodies
Chemical analysis

All Science Journal Classification (ASJC) codes

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

Cite this

A new approach of indirect enzyme-linked immunosorbent assay for determination of D-glutamic acid through in situ conjugation. / Sakamoto, Seiichi; Nagamitsu, Rika; Matsuura, Yurino; Tsuneura, Yumi; Kurose, Hitoshi; Tanaka, Hiroyuki; Morimoto, Satoshi.

In: Journal of Immunoassay and Immunochemistry, Vol. 37, No. 3, 03.05.2016, p. 296-306.

Research output: Contribution to journalArticle

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