Simultaneous monitoring of NanoLuc activity and DNA conjugation using agarose gel electrophoresis

Asfia Sultana; Anupriya M. Geethakumari; Kabir H. Biswas

doi:10.1016/j.ab.2026.116161

Simultaneous monitoring of NanoLuc activity and DNA conjugation using agarose gel electrophoresis

Asfia Sultana
, Anupriya M. Geethakumari
, Kabir H. Biswas^*

^*Corresponding author for this work

Biological and Biomedical Sciences

Hamad bin Khalifa University

Research output: Contribution to journal › Article › peer-review

Abstract

NanoLuc (NLuc) has been conjugated with DNA oligonucleotides (DNA oligo) for biosensing applications, which requires assessment of bioluminescence activity and conjugation efficiency. Here, we describe a straightforward, non-denaturing agarose gel electrophoresis–based assay that enables simultaneous monitoring of NLuc bioluminescence activity and NLuc-DNA oligo conjugation. Agarose gel electrophoresis of NLuc-DNA conjugation reaction samples and bioluminescence imaging revealed NLuc bioluminescence activity and a mobility shift of the protein confirming conjugation. Further, quantitative bioluminescence imaging allowed determination of active NLuc fraction and NLuc-DNA conjugation efficiency. The method reported here could be broadly applicable to other bioluminescent protein–DNA oligo conjugation workflows.

Original language	English
Article number	116161
Journal	Analytical Biochemistry
Volume	716
DOIs	https://doi.org/10.1016/j.ab.2026.116161
Publication status	Published - Sept 2026

Keywords

Agarose gel electrophoresis
Bioluminescence
NanoLuc
Protein-DNA oligo conjugation

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10.1016/j.ab.2026.116161

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@article{22c2570c5ff04717b73eb7e077478b54,

title = "Simultaneous monitoring of NanoLuc activity and DNA conjugation using agarose gel electrophoresis",

abstract = "NanoLuc (NLuc) has been conjugated with DNA oligonucleotides (DNA oligo) for biosensing applications, which requires assessment of bioluminescence activity and conjugation efficiency. Here, we describe a straightforward, non-denaturing agarose gel electrophoresis–based assay that enables simultaneous monitoring of NLuc bioluminescence activity and NLuc-DNA oligo conjugation. Agarose gel electrophoresis of NLuc-DNA conjugation reaction samples and bioluminescence imaging revealed NLuc bioluminescence activity and a mobility shift of the protein confirming conjugation. Further, quantitative bioluminescence imaging allowed determination of active NLuc fraction and NLuc-DNA conjugation efficiency. The method reported here could be broadly applicable to other bioluminescent protein–DNA oligo conjugation workflows.",

keywords = "Agarose gel electrophoresis, Bioluminescence, NanoLuc, Protein-DNA oligo conjugation",

author = "Asfia Sultana and Geethakumari, \{Anupriya M.\} and Biswas, \{Kabir H.\}",

note = "Publisher Copyright: {\textcopyright} 2026 Elsevier Inc.",

year = "2026",

month = sep,

doi = "10.1016/j.ab.2026.116161",

language = "English",

volume = "716",

journal = "Analytical Biochemistry",

issn = "0003-2697",

publisher = "Academic Press Inc.",

}

TY - JOUR

T1 - Simultaneous monitoring of NanoLuc activity and DNA conjugation using agarose gel electrophoresis

AU - Sultana, Asfia

AU - Geethakumari, Anupriya M.

AU - Biswas, Kabir H.

PY - 2026/9

Y1 - 2026/9

N2 - NanoLuc (NLuc) has been conjugated with DNA oligonucleotides (DNA oligo) for biosensing applications, which requires assessment of bioluminescence activity and conjugation efficiency. Here, we describe a straightforward, non-denaturing agarose gel electrophoresis–based assay that enables simultaneous monitoring of NLuc bioluminescence activity and NLuc-DNA oligo conjugation. Agarose gel electrophoresis of NLuc-DNA conjugation reaction samples and bioluminescence imaging revealed NLuc bioluminescence activity and a mobility shift of the protein confirming conjugation. Further, quantitative bioluminescence imaging allowed determination of active NLuc fraction and NLuc-DNA conjugation efficiency. The method reported here could be broadly applicable to other bioluminescent protein–DNA oligo conjugation workflows.

AB - NanoLuc (NLuc) has been conjugated with DNA oligonucleotides (DNA oligo) for biosensing applications, which requires assessment of bioluminescence activity and conjugation efficiency. Here, we describe a straightforward, non-denaturing agarose gel electrophoresis–based assay that enables simultaneous monitoring of NLuc bioluminescence activity and NLuc-DNA oligo conjugation. Agarose gel electrophoresis of NLuc-DNA conjugation reaction samples and bioluminescence imaging revealed NLuc bioluminescence activity and a mobility shift of the protein confirming conjugation. Further, quantitative bioluminescence imaging allowed determination of active NLuc fraction and NLuc-DNA conjugation efficiency. The method reported here could be broadly applicable to other bioluminescent protein–DNA oligo conjugation workflows.

KW - Agarose gel electrophoresis

KW - Bioluminescence

KW - NanoLuc

KW - Protein-DNA oligo conjugation

UR - https://www.scopus.com/pages/publications/105039981939

U2 - 10.1016/j.ab.2026.116161

DO - 10.1016/j.ab.2026.116161

M3 - Article

C2 - 42173336

AN - SCOPUS:105039981939

SN - 0003-2697

VL - 716

JO - Analytical Biochemistry

JF - Analytical Biochemistry

M1 - 116161

ER -

Simultaneous monitoring of NanoLuc activity and DNA conjugation using agarose gel electrophoresis

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Keywords

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