IJCEA 2025 Vol.16(1): 53-57
doi: 10.18178/ijcea.2025.16.1.839
Enhancement of a Humanized Nanobody against HER2 Tyrosine Kinase Production in the Cytoplasm of Escherichia coli
Manchusa Rungruangwong1, Nonth Submunkongtawee1, Warisara Kasemsukwimo1, Kwanpet Intasurat1, Tassaneewan Laksanasopin1, Lueacha Tabtimmai2, Kiattawee Choowongkomon3, and Dujduan Waraho-Zhmayev1,*
1. Faculty of Engineering, King Mongkut’s University of Technology Thonburi, Bangkok, 10140, Thailand
2. Department of Biotechnology, Faculty of Applied Science, King Mongkut’s University of Technology North Bangkok, Bangkok, 10800, Thailand
3. Department of Biochemistry, Faculty of Science, Kasetsart University, Bangkok, 10900, Thailand
Email: manchusakt111@gmail.com (M. R.); uunnonth@gmail.com (N. S.); warisara.wkm@gmail.com (W.K.); kwanpet.inta@gmail.com (K.I.); tassaneewan.lak@mail.kmutt.ac.th (T. L.); Lueacha.t@sci.kmutnb.ac.th (L.T.) fsciktc@ku.ac.th (K. C.); dujduan.war@mail.kmutt.ac.th (D. W.-Z.)
*Corresponding author
Manuscript received November 20, 2024; accepted March 1, 2025; published June 13, 2025
Abstract—Around 30% of invasive breast cancers are associated with the overexpression of the human epidermal growth factor receptor 2 (HER2) protein. While monoclonal antibodies are commonly used in targeted therapies, their large size hinders efficient delivery to tumor cells. Nanobodies, single-domain antibody fragments, provide a promising alternative due to their high binding affinity and small size. VHH17, a nanobody specific to HER2-Tyrosine Kinase (HER2-TK), blocks phosphorylation and reduces cancer cell proliferation. This study aimed to develop a humanized anti-HER2-TK nanobody and optimize its production in the cytoplasmic space of E. coli. The effects of temperature and induction on soluble expression were evaluated. Western blot analysis revealed that 20 °C and 1 mM IPTG induction were optimal for humanized VHH17 expression. These results demonstrate that E. coli can effectively express humanized VHH17 under proper conditions. Further optimization may enhance the yield of soluble VHH production.
Keywords—antibody fragment expression, Escherichia coli, human epidermal growth factor receptor 2 (HER2), tyrosine kinase, VHH single-domain antibody
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Cite: Manchusa Rungruangwong, Nonth Submunkongtawee, Warisara Kasemsukwimo, Kwanpet Intasurat, Tassaneewan Laksanasopin, Lueacha Tabtimmai, Kiattawee Choowongkomon, and Dujduan Waraho-Zhmayev, "Enhancement of a Humanized Nanobody against HER2 Tyrosine Kinase Production in the Cytoplasm of Escherichia coli," International Journal of Chemical Engineering and Applications vol. 16, no. 1, pp. 53-57, 2025.
