Generating Multispecific Antibodies Through Column-Based Redox Reactions: Part I

Area of Science:

• Biopharmaceutical manufacturing
• Protein chemistry
• Antibody engineering

Background:

• Multispecific antibodies are crucial for addressing unmet patient needs in the pharmaceutical industry.
• Current manufacturing of electrostatic-steering multispecific antibodies involves complex processing steps.
• There is a need for simplified and efficient methods for producing these complex biologics.

Purpose of the Study:

• To develop and characterize a novel, simplified redox process for generating multispecific antibodies via electrostatic steering.
• To evaluate the efficiency and product quality of this new manufacturing strategy.
• To demonstrate the applicability of the method across diverse multispecific antibody formats.

Main Methods:

• Simultaneous binding of two parental homodimer antibodies to Protein A chromatography resin.
• Column-based reduction of interchain disulfide bonds using a reductant wash.
• Elution, neutralization, and subsequent oxidation to form the intact heterodimer multispecific antibody.
• Characterization of reduction, heterodimerization, and oxidation kinetics and mechanisms.

Main Results:

• Successful demonstration of the novel redox process for five distinct multispecific antibodies with varied specificities and IgG subclasses.
• Characterization confirmed efficient conversion and acceptable product quality.
• The new strategy significantly simplifies downstream purification compared to traditional methods.

Conclusions:

• The developed redox process offers a simplified and potentially more cost-effective approach to manufacturing multispecific antibodies.
• This method maintains product quality and yield, making it suitable for pharmaceutical bioprocesses.
• Implementation can reduce manufacturing complexity for electrostatic-steering based multispecific molecules.