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Lysibody-induced Phagocytosis Determination Assay

With advanced platforms and extensive knowledge, Creative Biolabs provides comprehensive lysosome-induced phagocytosis assay services, from experimental design to final data analysis, to facilitate your great success in the field.

Overview of Lysibody-induced Phagocytosis

In recent years, lysibody technology has emerged as an alternative method to generate promising opsonized antibodies against bacterial surface carbohydrates. Lysibodies are defined as chimeras between the Fc region of a human immunoglobulin G (IgG) antibody and carbohydrate derived from a cell wall hydrolase. Like normal IgG antibodies, lysibodies have many effector functions and exhibit robust complement fixation and induction of phagocytosis. Induction of phagocytosis is beneficial for potentially clearing the infection, reducing the risk of cytokine release, as no bacterial lysate is released. In addition, lysibodies play a crucial role in protecting mice from multiple bacteria attacks, especially lethal methicillin-resistant Staphylococcus aureus (MRSA).

Assays for Lysibody-induced Phagocytosis

  • Principle

Destruction of bacteria by phagocytes is a critical step in the control of staphylococcal infection. To further explore the extent of lysibody-induced phagocytosis, cells of the bacterial strain expressing fluorescent proteins are incubated with monolayers of macrophages or neutrophils in the presence of various lysibodies. Phagocytes are washed and analyzed by flow cytometry (FC) and microscopy to confirm the degree of phagocytosis.

  • Macrophages phagocytosis assay

Different assays are used to explore the role of lysibodies inducing macrophage phagocytosis and killing bacteria. A typical experimental procedure is as follows: naive macrophages or peritoneal murine macrophages are incubated with a green fluorescent protein (GFP)-labeled bacteria in the presence of various concentrations of lysibodies; the degree of phagocytosis is further confirmed by the determination of percent phagocytosis by FC after the removal of macrophages.

AtlA lysibody induces phagocytosis of S. aureus by macrophages.Fig.1 AtlA lysibody induces phagocytosis of S. aureus by macrophages. (Raz, 2017)

  • Neutrophils phagocytosis assay

The assay of lysibodies to induce neutrophil phagocytosis of bacteria is equivalent to the macrophage assay described above. Typically, neutrophils are incubated with fluorescein isothiocyanate (FITC)-labeled bacterial strains in the presence of lysibodies and an appropriate concentration of complement. The effect of different lysibodies on the phagocytosis of bacteria by neutrophils is further confirmed by FC to determine the percentage of phagocytosis.

What Can We Do?

As an expert in the field of lysibody research, Creative Biolabs has extensive experience in exploring the function of lysibody. With advanced experimental platforms and strong expertise, our dedicated scientists focus on providing a variety of assays to assist the progress of your lysibodies-induced phagocytosis project. Many techniques are available at Creative Biolabs, such as FC, fluorescent labeling, microscopy, and enzyme-linked immunosorbent assay (ELISA). Based on our rigorous experimental design, we are confident to provide reproducible experimental results in short cycle times.

At Creative Biolabs, assays for lysibody-induced phagocytosis are performed under a strictly controlled process to ensure complete customer satisfaction. We will provide reasonable and applicable solutions and detection methods according to your project needs. If you are interested in our services, please feel free to contact us for more details.

Reference:

  1. Raz, A.; et al. Lysibodies are IgG Fc fusions with lysin binding domains targeting Staphylococcus aureus wall carbohydrates for effective phagocytosis. Proceedings of the National Academy of Sciences. 2017, 114(18): 4781-4786.
For Research Use Only. Do NOT use in humans.

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