Highly robust and optimized conjugation of antibodies to nanoparticles using quantitatively validated protocols.
Antibody-conjugated nanoparticles (NPs) have attracted great attention in diagnostic and therapeutic applications due to their high sensitivity and specificity for biotargets, as well as their wide applicability. Unfortunately, these features are significantly affected by antibody conjugation methods in terms of conjugation efficiency, orientation of the target binding site in the antibody, and denaturation during chemical conjugation reactions. Furthermore, the number of conjugated antibodies on each NP and the overall targeting efficacy are critical factors for a quantitative bioassay with antibody-conjugated NPs. Herein, we report a versatile and oriented antibody conjugation method using copper-free click chemistry. Moreover, the number of conjugated antibodies and their binding capacity were quantitatively and experimentally evaluated using fluorescently-labeled antibodies and antigens. The strong binding capability of antibody-conjugated NPs prepared using the copper-free click chemistry-based conjugation strategy was 8 times superior to the binding capability seen following the use of the EDC/NHS-coupling method. Additionally, the versatility of the developed antibody conjugation method was also demonstrated by conjugation of the antibody to three kinds of silica-encapsulated NPs.
Department of Chemistry Education, Seoul National University, Seoul 08826, Korea. jeongdh@snu.ac.kr.
Sinyoung
S
Jeong
Department of Nuclear Medicine, Seoul National University College of Medicine, Seoul 03080, Korea. wonza43@snu.ac.kr and Major in Biomedical Sciences, Department of Biomedical Sciences, Seoul National University Graduate School, Seoul 08826, Korea.
Ji Yong
JY
Park
Department of Chemistry Education, Seoul National University, Seoul 08826, Korea. jeongdh@snu.ac.kr.
Myeong Geun
MG
Cha
Department of Chemistry Education, Seoul National University, Seoul 08826, Korea. jeongdh@snu.ac.kr.
Hyejin
H
Chang
Department of Nuclear Medicine, Seoul National University College of Medicine, Seoul 03080, Korea. wonza43@snu.ac.kr.
Yong-Il
YI
Kim
Department of Bioscience and Biotechnology, Konkuk University, Seoul 05029, Korea.
Hyung-Mo
HM
Kim
Department of Bioscience and Biotechnology, Konkuk University, Seoul 05029, Korea.
Bong-Hyun
BH
Jun
Department of Nuclear Medicine, Seoul National University College of Medicine, Seoul 03080, Korea. wonza43@snu.ac.kr and Department of Molecular Medicine and Biopharmaceutical Sciences, Graduate School of Convergence Science and Technology, and College of Medicine or College of Pharmacy, Seoul National University, Seoul 08826, Korea.
Dong Soo
DS
Lee
School of Chemical and Biological Engineering, Seoul National University, Seoul 08826, Korea.
Yoon-Sik
YS
Lee
Department of Nuclear Medicine, Seoul National University College of Medicine, Seoul 03080, Korea. wonza43@snu.ac.kr.
Jae Min
JM
Jeong
Department of Nuclear Medicine, Seoul National University College of Medicine, Seoul 03080, Korea. wonza43@snu.ac.kr and Department of Molecular Medicine and Biopharmaceutical Sciences, Graduate School of Convergence Science and Technology, and College of Medicine or College of Pharmacy, Seoul National University, Seoul 08826, Korea.
Yun-Sang
YS
Lee
Department of Chemistry Education, Seoul National University, Seoul 08826, Korea. jeongdh@snu.ac.kr.
Dae Hong
http://orcid.org/0000-0001-9407-7006
DH
Jeong
D000906:Antibodies / Q000737:chemistry*; D057930:Click Chemistry; D018796:Immunoconjugates / Q000737:chemistry*; D053758:Nanoparticles / Q000737:chemistry*; D012822:Silicon Dioxide
D000906:Antibodies; D018796:Immunoconjugates; D012822:Silicon Dioxide