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Optimizing assembly and production of native bispecific antibodies by codon de-optimization
Giovanni Magistrelli1, Yves Poitevin1, Florence Schlosser1
1a Novimmune SA , Plan-les-Ouates , Geneva , Switzerland.
Mabs
|December 22, 2016
Summary
Optimizing bispecific antibody production involves balancing polypeptide chain expression. De-optimizing lambda chain expression unexpectedly doubled the yield of kappa-lambda (κλ) bodies by improving assembly.
Area of Science:
- Biotechnology
- Protein Engineering
- Immunology
Background:
- Bispecific antibody production can be limited by uneven expression of multiple polypeptide chains.
- Kappa-lambda (κλ) bodies are bispecific antibodies with a native IgG structure, comprising a common heavy chain and two distinct light chains (kappa and lambda).
- Unforced chain pairing leads to co-secretion of both monospecific and bispecific antibodies, necessitating stoichiometric expression for maximal bispecific yield.
Purpose of the Study:
- To investigate strategies for improving the yield of κλ bodies, specifically addressing suboptimal production due to low kappa chain expression.
- To evaluate the impact of codon optimization and de-optimization on the expression levels of individual light chains and overall bispecific antibody assembly.
- To understand the relationship between polypeptide chain expression stoichiometry and the efficient assembly of complex recombinant proteins.
Main Methods:
- Selection of a κλ body exhibiting suboptimal yield attributed to low kappa chain expression.
- Application of codon optimization techniques to the kappa chain to enhance its expression.
- Systematic introduction of non-optimal codons into the lambda chain sequence to modulate its expression level.
Main Results:
- Codon optimization of the kappa chain failed to improve the overall bispecific antibody yield.
- Progressive codon de-optimization of the lambda chain led to a decrease in its expression.
- The de-optimization of lambda chain expression resulted in an improved ratio of bispecific to monospecific antibodies, effectively doubling the κλ body yield.
Conclusions:
- Recombinant complex assembly is an interconnected process influenced by the relative expression levels of constituent polypeptide chains.
- Reducing the expression of one polypeptide chain can paradoxically enhance the overall yield of the desired complex.
- Strategic codon de-optimization offers a viable approach to optimize the assembly and yield of complex bispecific antibodies like κλ bodies.
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