Bacterial Lipid Modification of ICP11 and a New ELISA System Applicable for WSSV Infection Detection

Area of Science:

• Biotechnology
• Protein Engineering
• Immunodiagnostics

Background:

• Hydrophilic protein immobilization on hydrophobic surfaces is a challenge in ELISA.
• Current methods lack stability and efficiency for certain protein targets.

Purpose of the Study:

• To develop a novel protein engineering strategy for stable protein immobilization in ELISA.
• To create a sensitive diagnostic tool for White Spot Syndrome Virus (WSSV) in shrimp farming.

Main Methods:

• Recombinant bacterial lipid modification of proteins using N-acyl-S-diacylglyceryl Cys in E. coli.
• Lipid-modified ICP11 protein from WSSV used as a target in a new ELISA.
• Competitive ELISA format developed for antigen detection in shrimp tissues.

Main Results:

• Lipid modification enabled stable immobilization of hydrophilic proteins on polystyrene surfaces.
• Lipid modification acted as an adjuvant, enhancing antibody titers 16-fold.
• The competitive ELISA achieved a detection limit of 250 pg for ICP11 protein.

Conclusions:

• Bacterial lipid modification is an effective post-translational engineering strategy for protein immobilization and enhancing immune response.
• The developed ELISA offers a sensitive and efficient method for WSSV diagnosis in aquaculture.
• This approach has potential applications in other diagnostic assays requiring stable protein immobilization.