A new approach of indirect enzyme-linked immunosorbent assay for determination of D-glutamic acid through in situ conjugation. Issue 3 (3rd May 2016)

• Record Type: Journal Article
• Authors: Sakamoto, Seiichi; Nagamitsu, Rika; Matsuura, Yurino; Tsuneura, Yumi; Kurose, Hitoshi; Tanaka, Hiroyuki; Morimoto, Satoshi
• Is Part Of: Journal of immunoassay & immunochemistry. Volume 37:Issue 3(2016)
• Page Start: 296
• Page End: 306
• Publication Date: 2016-05-03
• Subjects: D-glutamic acid (D-Glu) -- enzyme-linked immunosorbent assay (ELISA) -- glutaraldehyde (GA) -- in situ conjugation -- monoclonal antibody (MAb); Immunoassay -- Periodicals; Immunochemistry -- Periodicals; Immunoassay -- periodicals; Immunochemistry -- periodicals; 571.96405
• DOI: 10.1080/15321819.2015.1135161
• Languages: English
• ISSNs: 1532-1819

Abstract

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 7.81μ 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.