TY - JOUR
T1 - Direct conjugation of streptavidin to encoded hydrogel microparticles for multiplex biomolecule detection with rapid probe-set modification
AU - Roh, Yoon Ho
AU - Kim, Ju Yeon
AU - Mun, Seok Joon
AU - Lee, Hye Sun
AU - Hwang, Changhyun
AU - Park, Kyong Hwa
AU - Bong, Ki Wan
N1 - Funding Information:
Funding: This work has been supported by the Engineering Research Center of Excellence Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Science, ICT & Future Planning (NRF-2016R1A5A1010148), the Basic Science Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Education (NRF-2018R1D1A1B07046577), and the grant from the Next-Generation Biogreen 21 Program (No. PJ013158), Rural Development Administration, Republic of Korea.
Publisher Copyright:
© 2020 by the authors.
PY - 2020/3/1
Y1 - 2020/3/1
N2 - Encoded hydrogel microparticles synthesized via flow lithography have drawn attention for multiplex biomarker detection due to their high multiplex capability and solution-like hybridization kinetics. However, the current methods for preparing particles cannot achieve a flexible, rapid probe-set modification, which is necessary for the production of various combinations of target panels in clinical diagnosis. In order to accomplish the unmet needs, streptavidin was incorporated into the encoded hydrogel microparticles to take advantage of the rapid streptavidin-biotin interactions that can be used in probe-set modification. However, the existing methods suffer from low efficiency of streptavidin conjugation, cause undesirable deformation of particles, and impair the assay capability. Here, we present a simple and powerful method to conjugate streptavidin to the encoded hydrogel microparticles for better assay performance and rapid probe-set modification. Streptavidin was directly conjugated to the encoded hydrogel microparticles using the aza-Michael addition click reaction, which can proceed in mild, aqueous condition without catalysts. A highly flexible and sensitive assay was developed to quantify DNA and proteins using streptavidin-conjugated encoded hydrogel microparticles. We also validated the potential applications of our particles conducting multiplex detection of cancer-related miRNAs.
AB - Encoded hydrogel microparticles synthesized via flow lithography have drawn attention for multiplex biomarker detection due to their high multiplex capability and solution-like hybridization kinetics. However, the current methods for preparing particles cannot achieve a flexible, rapid probe-set modification, which is necessary for the production of various combinations of target panels in clinical diagnosis. In order to accomplish the unmet needs, streptavidin was incorporated into the encoded hydrogel microparticles to take advantage of the rapid streptavidin-biotin interactions that can be used in probe-set modification. However, the existing methods suffer from low efficiency of streptavidin conjugation, cause undesirable deformation of particles, and impair the assay capability. Here, we present a simple and powerful method to conjugate streptavidin to the encoded hydrogel microparticles for better assay performance and rapid probe-set modification. Streptavidin was directly conjugated to the encoded hydrogel microparticles using the aza-Michael addition click reaction, which can proceed in mild, aqueous condition without catalysts. A highly flexible and sensitive assay was developed to quantify DNA and proteins using streptavidin-conjugated encoded hydrogel microparticles. We also validated the potential applications of our particles conducting multiplex detection of cancer-related miRNAs.
KW - Aza-Michael addition click reaction
KW - Encoded hydrogel microparticle
KW - Probe-set modification
KW - Stop flow lithography
KW - Streptavidin
UR - http://www.scopus.com/inward/record.url?scp=85082704231&partnerID=8YFLogxK
U2 - 10.3390/polym12030546
DO - 10.3390/polym12030546
M3 - Article
AN - SCOPUS:85082704231
VL - 12
JO - Polymers
JF - Polymers
SN - 2073-4360
IS - 3
M1 - 546
ER -