Identification of two neutralizing human single-chain variable fragment antibodies targeting Staphylococcus aureus alpha-hemolysin

Publish Year: 1401
نوع سند: مقاله ژورنالی
زبان: English
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JR_IJBMS-25-10_007

تاریخ نمایه سازی: 11 مهر 1401

Abstract:

Objective(s): The inability of the host immune system to defeat Staphylococcus aureus is due to various secreted virulent factors such as leukocidins, superantigens, and hemolysins, which interrupt the function of immune components. Alpha-hemolysin is one of the most studied cytolysins due to its pronounced effect on developing staphylococcal infections. Alpha-hemolysin-neutralizing antibodies are among the best candidates for blocking the toxin activity and preventing S. aureus pathogenesis. Materials and Methods: A human single-chain variable fragment (scFv) phage display library was biopanned against alpha-hemolysin. The selected phage clones were assessed based on their binding ability to alpha-hemolysin. The binding specificity and affinity of two scFvs (designated SP۱۹۲ and SP۲۲۰) to alpha-hemolysin were determined by enzyme-linked immunosorbent assay. Furthermore, the neutralizing activity of SP۱۹۲ and SP۲۲۰ was examined by concurrent incubation of rabbit red blood cells (RBCs) with alpha-hemolysin and scFvs.Results: SP۱۹۲ and SP۲۲۰ showed significant binding to alpha-hemolysin compared with the control proteins, including bovine serum albumin, human adiponectin, and toxic shock syndrome toxin-۱. Besides, both scFvs showed high-affinity binding to alpha-hemolysin in the nanomolar range (Kaff: ۰.۹ and ۰.۷ nM-۱, respectively), leading to marked inhibition of alpha-hemolysin-mediated lysis of rabbit RBCs (۷۳% and ۸۴% inhibition; respectively). Conclusion: SP۱۹۲ and SP۲۲۰ scFvs can potentially be used as alpha-hemolysin-neutralizing agents in conjunction with conventional antibiotics to combat S. aureus infections. 

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Authors

Somayeh Piri-Gavgani

Department of Mycobacteriology and Pulmonary Research, Pasteur Institute of Iran, Tehran, Iran

Mostafa Ghanei

Chemical Injuries Research Center, Baqiyatallah University of Medical Sciences, Tehran, Iran

Abolfazl Fateh

Department of Mycobacteriology and Pulmonary Research, Pasteur Institute of Iran, Tehran, Iran

Seyed Davar Siadat

Department of Mycobacteriology and Pulmonary Research, Pasteur Institute of Iran, Tehran, Iran

Leila Nematollahi

Biotechnology Research Center, Pasteur Institute of Iran, Tehran, Iran

Fatemeh Rahimi-Jamnani

Department of Mycobacteriology and Pulmonary Research, Pasteur Institute of Iran, Tehran, Iran

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