TY - JOUR
T1 - A new approach of indirect enzyme-linked immunosorbent assay for determination of D-glutamic acid through in situ conjugation
AU - Sakamoto, Seiichi
AU - Nagamitsu, Rika
AU - Matsuura, Yurino
AU - Tsuneura, Yumi
AU - Kurose, Hitoshi
AU - Tanaka, Hiroyuki
AU - Morimoto, Satoshi
N1 - Publisher Copyright:
Copyright © Taylor & Francis Group, LLC.
Copyright:
Copyright 2017 Elsevier B.V., All rights reserved.
PY - 2016/5/3
Y1 - 2016/5/3
N2 - 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.
AB - 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.
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U2 - 10.1080/15321819.2015.1135161
DO - 10.1080/15321819.2015.1135161
M3 - Article
C2 - 26744787
AN - SCOPUS:84962636305
SN - 1532-1819
VL - 37
SP - 296
EP - 306
JO - Journal of Immunoassay and Immunochemistry
JF - Journal of Immunoassay and Immunochemistry
IS - 3
ER -