Selection of antibodies for immunoaffinity chromatography

Area of Science:

• Biochemistry
• Immunology
• Chromatography

Background:

• Immunoaffinity chromatography offers choices between polyclonal and monoclonal antibodies, and immobilized antigens or antibodies.
• Historically, antibody selection lacked rational principles, relying on available animal sera.
• Monoclonal antibodies have expanded choices but necessitate systematic approaches for their production and application.

Purpose of the Study:

• To focus on monoclonal antibody selection for immunoaffinity chromatography.
• To highlight the need for rational principles in antibody production, selection, and use.
• To emphasize the importance of assay development for efficient antibody selection.

Main Methods:

• Concentrating on the selection process for monoclonal antibodies.
• Considering aspects like immunization, fusion, cloning, and solid-phase preparation.
• Utilizing automated or semi-automated assay techniques such as enzyme-linked immunosorbent assay (ELISA), radioimmunoassay (RIA), and immunoradiometric assay (IRMA).

Main Results:

• Monoclonal antibodies offer broader, more rational choices compared to polyclonal antibodies.
• Systematic approaches and principles are crucial for effective antibody selection and use.
• Automated assays significantly reduce the tedium of empirical testing in antibody selection.

Conclusions:

• Rational antibody selection, particularly with monoclonal antibodies, is vital for successful immunoaffinity chromatography.
• Integrated control over immunization, antibody production, and assay development is essential.
• Automated assay techniques are fundamental to efficient and reliable antibody selection processes.