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

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

  • Bioconjug Chem. 2022 Nov 16;33(11):2180-2188. doi: 10.1021/acs.bioconjchem.2c00416.
Chiao-Yu Hsiao 1,  Jun-Lun Meng 1,  Jung-Zhe Hong 1,  Xuan-Huong Ly 1,  Meng-Hsuan Lin 1,  Chin-Yuan Chang 1 2,  Minh-Tram T Nguyen 1,  Tian-Lu Cheng 2,  Wen-Wei Lin 3,  Pierre-Alain Burnouf 4,  Talal Salem Al-Qaisi 5,  En-Shuo Liu 6,  Yu-Cheng Su 1 2
Affiliations

Affiliations

  • 1 Department of Biological Science and Technology, Center for Intelligent Drug Systems and Smart Bio-devices (IDS2B), National Yang Ming Chiao Tung University, Hsinchu 300193, Taiwan.
  • 2 Department of Biomedical Science and Environmental Biology, Drug Development and Value Creation Research Center, Kaohsiung Medical University, Kaohsiung 807378, Taiwan.
  • 3 School of Post-Baccalaureate Medicine, College of Medicine, Kaohsiung Medical University, Kaohsiung 807378, Taiwan.
  • 4 International Center for Wound Repair and Regeneration (iWRR), National Cheng-Kung University, Tainan 701401, Taiwan.
  • 5 Department of Medical Laboratory Sciences, Pharmacological and Diagnostic Research Centre, Faculty of Allied Medical Sciences, Al-Ahliyya Amman University, Amman 19328, Jordan.
  • 6 Graduate Institute of Medicine, College of Medicine, Kaohsiung Medical University, Kaohsiung 807378, Taiwan.
PMID: 36320124 DOI: 10.1021/acs.bioconjchem.2c00416
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 mPEG5K and high-molecular-weight (hmw) mPEG20K at 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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