CLC number: R782.2
On-line Access: 2017-10-05
Received: 2016-08-11
Revision Accepted: 2017-02-19
Crosschecked: 2017-09-13
Cited: 0
Clicked: 5307
Rui Wang, Yuan Chen, Kai Fan, Feng Ji, Jian Wu, Yong-hua Yu. Nominal effective immunoreaction volume of magnetic beads at single bead level[J]. Journal of Zhejiang University Science B, 2017, 18(10): 845-853.
@article{title="Nominal effective immunoreaction volume of magnetic beads at single bead level",
author="Rui Wang, Yuan Chen, Kai Fan, Feng Ji, Jian Wu, Yong-hua Yu",
journal="Journal of Zhejiang University Science B",
volume="18",
number="10",
pages="845-853",
year="2017",
publisher="Zhejiang University Press & Springer",
doi="10.1631/jzus.B1600358"
}
%0 Journal Article
%T Nominal effective immunoreaction volume of magnetic beads at single bead level
%A Rui Wang
%A Yuan Chen
%A Kai Fan
%A Feng Ji
%A Jian Wu
%A Yong-hua Yu
%J Journal of Zhejiang University SCIENCE B
%V 18
%N 10
%P 845-853
%@ 1673-1581
%D 2017
%I Zhejiang University Press & Springer
%DOI 10.1631/jzus.B1600358
TY - JOUR
T1 - Nominal effective immunoreaction volume of magnetic beads at single bead level
A1 - Rui Wang
A1 - Yuan Chen
A1 - Kai Fan
A1 - Feng Ji
A1 - Jian Wu
A1 - Yong-hua Yu
J0 - Journal of Zhejiang University Science B
VL - 18
IS - 10
SP - 845
EP - 853
%@ 1673-1581
Y1 - 2017
PB - Zhejiang University Press & Springer
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DOI - 10.1631/jzus.B1600358
Abstract: immunomagnetic bead (IMB)-based enzyme-linked immunosorbent assay (ELISA) has been the tool frequently used for protein detection in research and clinical laboratories. For most ELISA reactions the recommended dosage of IMBs is usually according to their weight (mg) or mass fraction (w/v) instead of the bead number. Consequently, the processes occurring in the immediate vicinity of the IMBs have always been ignored by researchers and they cannot be revealed in detail during the ELISA reaction. In this paper, we established the relationship between number of IMBs and colorimetric results, and further proposed a new concept of “nominal effective immunoreaction volume (NEIV)” to characterize a single IMB during ELISA reaction. Results showed that the NEIV of a single IMB has a constant value, which is unrelated to the amount of beads and the concentration of antigen. Optimal results of the colorimetric ELISA are achieved when the incubation volume meets each IMB’s NEIV and is no longer enhanced by increasing the incubation volume. Thus, the reliable and relatively precise number of IMBs for ELISA detection during practical application could be determined. Most importantly, a study using IMB’s NEIV would lay the foundation for a kinetics analysis of IMBs and antigens for future study.
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[22]List of electronic supplementary materials
[23]Fig. S1 Principle of EDC-NHS method for antibody modification of magnetic beads
[24]Fig. S2 Principle of IMB-based ELISA assay for PSA antigen detection
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