Engineering a High-Affinity Anti-Methoxy Poly(ethylene glycol) (mPEG) Antibody for Sensitive Immunosensing of mPEGylated Therapeutics and mPEG Molecules

Chiao Yu Hsiao; Jun Lun Meng; Jung Zhe Hong; Xuan Huong Ly; Meng Hsuan Lin; Chin Yuan Chang; Minh Tram T. Nguyen; Tian Lu Cheng; Wen Wei Lin; Pierre Alain Burnouf; Talal Salem Al-Qaisi; En Shuo Liu; Yu Cheng Su

doi:10.1021/acs.bioconjchem.2c00416

Engineering a High-Affinity Anti-Methoxy Poly(ethylene glycol) (mPEG) Antibody for Sensitive Immunosensing of mPEGylated Therapeutics and mPEG Molecules

Chiao Yu Hsiao, Jun Lun Meng, Jung Zhe Hong, Xuan Huong Ly, Meng Hsuan Lin, Chin Yuan Chang, Minh Tram T. Nguyen, Tian Lu Cheng, Wen Wei Lin, Pierre Alain Burnouf, Talal Salem Al-Qaisi, En Shuo Liu, Yu Cheng Su^*

^*Corresponding author for this work

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

2 Scopus citations

Abstract

Sensitive quantification of methoxy poly(ethylene glycol) (mPEG)-conjugated therapeutics for pharmacokinetic determination is critical for mPEGylated drug development. However, sensitive measurement of low-molecular-weight (lmw) mPEG compounds remains challenging due to epitope competition between backbone-specific anti-PEG antibodies. Here, we engineered a high-affinity methoxy-specific anti-mPEG antibody for sensitive quantification of free mPEG molecules and mPEGylated therapeutics. The affinity-enhanced h15-2Y antibody variant shows a 10.3-fold increase in mPEG-binding activity compared to parental h15-2b. h15-2Y-based sandwich ELISA can effectively quantify lmw mPEG_5Kand high-molecular-weight (hmw) mPEG_20Kat concentrations as low as 3.4 and 5.1 ng mL^-1, respectively. Moreover, lmw mPEG compounds (560, 750, 1000, and 2000 Da) can be efficiently quantified via h15-2Y-based competitive ELISA with detection limits at nanomolar levels. This study provides a promising approach for application in the quantitative analysis of the various sizes of mPEG molecules to accelerate the timeline of mPEG-conjugated drug development.

Original language	English
Pages (from-to)	2180-2188
Number of pages	9
Journal	Bioconjugate Chemistry
Volume	33
Issue number	11
DOIs	https://doi.org/10.1021/acs.bioconjchem.2c00416
State	Published - 16 Nov 2022

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Hsiao, C. Y., Meng, J. L., Hong, J. Z., Ly, X. H., Lin, M. H., Chang, C. Y., Nguyen, M. T. T., Cheng, T. L., Lin, W. W., Burnouf, P. A., Al-Qaisi, T. S., Liu, E. S., & Su, Y. C. (2022). Engineering a High-Affinity Anti-Methoxy Poly(ethylene glycol) (mPEG) Antibody for Sensitive Immunosensing of mPEGylated Therapeutics and mPEG Molecules. Bioconjugate Chemistry, 33(11), 2180-2188. https://doi.org/10.1021/acs.bioconjchem.2c00416

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title = "Engineering a High-Affinity Anti-Methoxy Poly(ethylene glycol) (mPEG) Antibody for Sensitive Immunosensing of mPEGylated Therapeutics and mPEG Molecules",

abstract = "Sensitive quantification of methoxy poly(ethylene glycol) (mPEG)-conjugated therapeutics for pharmacokinetic determination is critical for mPEGylated drug development. However, sensitive measurement of low-molecular-weight (lmw) mPEG compounds remains challenging due to epitope competition between backbone-specific anti-PEG antibodies. Here, we engineered a high-affinity methoxy-specific anti-mPEG antibody for sensitive quantification of free mPEG molecules and mPEGylated therapeutics. The affinity-enhanced h15-2Y antibody variant shows a 10.3-fold increase in mPEG-binding activity compared to parental h15-2b. h15-2Y-based sandwich ELISA can effectively quantify lmw mPEG5Kand high-molecular-weight (hmw) mPEG20Kat concentrations as low as 3.4 and 5.1 ng mL-1, respectively. Moreover, lmw mPEG compounds (560, 750, 1000, and 2000 Da) can be efficiently quantified via h15-2Y-based competitive ELISA with detection limits at nanomolar levels. This study provides a promising approach for application in the quantitative analysis of the various sizes of mPEG molecules to accelerate the timeline of mPEG-conjugated drug development.",

author = "Hsiao, {Chiao Yu} and Meng, {Jun Lun} and Hong, {Jung Zhe} and Ly, {Xuan Huong} and Lin, {Meng Hsuan} and Chang, {Chin Yuan} and Nguyen, {Minh Tram T.} and Cheng, {Tian Lu} and Lin, {Wen Wei} and Burnouf, {Pierre Alain} and Al-Qaisi, {Talal Salem} and Liu, {En Shuo} and Su, {Yu Cheng}",

year = "2022",

month = nov,

day = "16",

doi = "10.1021/acs.bioconjchem.2c00416",

language = "English",

volume = "33",

pages = "2180--2188",

journal = "Bioconjugate Chemistry",

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publisher = "American Chemical Society",

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Hsiao, CY, Meng, JL, Hong, JZ, Ly, XH, Lin, MH, Chang, CY, Nguyen, MTT, Cheng, TL, Lin, WW, Burnouf, PA, Al-Qaisi, TS, Liu, ES & Su, YC 2022, 'Engineering a High-Affinity Anti-Methoxy Poly(ethylene glycol) (mPEG) Antibody for Sensitive Immunosensing of mPEGylated Therapeutics and mPEG Molecules', Bioconjugate Chemistry, vol. 33, no. 11, pp. 2180-2188. https://doi.org/10.1021/acs.bioconjchem.2c00416

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T1 - Engineering a High-Affinity Anti-Methoxy Poly(ethylene glycol) (mPEG) Antibody for Sensitive Immunosensing of mPEGylated Therapeutics and mPEG Molecules

AU - Hsiao, Chiao Yu

AU - Meng, Jun Lun

AU - Hong, Jung Zhe

AU - Ly, Xuan Huong

AU - Lin, Meng Hsuan

AU - Chang, Chin Yuan

AU - Nguyen, Minh Tram T.

AU - Cheng, Tian Lu

AU - Lin, Wen Wei

AU - Burnouf, Pierre Alain

AU - Al-Qaisi, Talal Salem

AU - Liu, En Shuo

AU - Su, Yu Cheng

PY - 2022/11/16

Y1 - 2022/11/16

N2 - Sensitive quantification of methoxy poly(ethylene glycol) (mPEG)-conjugated therapeutics for pharmacokinetic determination is critical for mPEGylated drug development. However, sensitive measurement of low-molecular-weight (lmw) mPEG compounds remains challenging due to epitope competition between backbone-specific anti-PEG antibodies. Here, we engineered a high-affinity methoxy-specific anti-mPEG antibody for sensitive quantification of free mPEG molecules and mPEGylated therapeutics. The affinity-enhanced h15-2Y antibody variant shows a 10.3-fold increase in mPEG-binding activity compared to parental h15-2b. h15-2Y-based sandwich ELISA can effectively quantify lmw mPEG5Kand high-molecular-weight (hmw) mPEG20Kat concentrations as low as 3.4 and 5.1 ng mL-1, respectively. Moreover, lmw mPEG compounds (560, 750, 1000, and 2000 Da) can be efficiently quantified via h15-2Y-based competitive ELISA with detection limits at nanomolar levels. This study provides a promising approach for application in the quantitative analysis of the various sizes of mPEG molecules to accelerate the timeline of mPEG-conjugated drug development.

AB - Sensitive quantification of methoxy poly(ethylene glycol) (mPEG)-conjugated therapeutics for pharmacokinetic determination is critical for mPEGylated drug development. However, sensitive measurement of low-molecular-weight (lmw) mPEG compounds remains challenging due to epitope competition between backbone-specific anti-PEG antibodies. Here, we engineered a high-affinity methoxy-specific anti-mPEG antibody for sensitive quantification of free mPEG molecules and mPEGylated therapeutics. The affinity-enhanced h15-2Y antibody variant shows a 10.3-fold increase in mPEG-binding activity compared to parental h15-2b. h15-2Y-based sandwich ELISA can effectively quantify lmw mPEG5Kand high-molecular-weight (hmw) mPEG20Kat concentrations as low as 3.4 and 5.1 ng mL-1, respectively. Moreover, lmw mPEG compounds (560, 750, 1000, and 2000 Da) can be efficiently quantified via h15-2Y-based competitive ELISA with detection limits at nanomolar levels. This study provides a promising approach for application in the quantitative analysis of the various sizes of mPEG molecules to accelerate the timeline of mPEG-conjugated drug development.

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AN - SCOPUS:85141640265

SN - 1043-1802

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JO - Bioconjugate Chemistry

JF - Bioconjugate Chemistry

IS - 11

ER -

Engineering a High-Affinity Anti-Methoxy Poly(ethylene glycol) (mPEG) Antibody for Sensitive Immunosensing of mPEGylated Therapeutics and mPEG Molecules

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