Development of a mammalian cell-based ZZ display system for IgG quantification

Area of Science:

• Biotechnology
• Immunology
• Cell Biology

Background:

• Developing effective antibody detection methods is crucial for biological laboratories and antibody development companies.
• Current methods may lack convenience and versatility for detecting highly active antibodies.

Purpose of the Study:

• To establish a mammalian cell-based ZZ display system for efficient antibody quantification.
• To develop a versatile platform for detecting and quantifying immunoglobulin G (IgG) from various species.

Main Methods:

• Construction and transfection of a eukaryotic ZZ-displayed plasmid into CHO cells.
• Screening and selection of stable ZZ display cells (CHO-ZZ cells) using flow cytometry.
• Incubation of CHO-ZZ cells with IgG samples and analysis of relative fluorescence intensity for quantification.

Main Results:

• A highly efficient ZZ display system (CHO-ZZ cells) was established, achieving near 100% ZZ peptide display.
• CHO-ZZ cells demonstrated the ability to capture diverse IgG types, including human, rabbit, donkey, mouse, and goat.
• The system enabled quantification of human IgG within a range of 12.5-1000 ng/mL and facilitated identification of high-yielding monoclonal cell lines.

Conclusions:

• A highly efficient CHO-ZZ display system for IgG quantification under physiological conditions has been developed.
• This system eliminates the need for antibody purification, streamlining the antibody development process.
• The established system offers a versatile and convenient method for antibody detection and quantification.