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Improving Multispecific Antibody Bioprocesses Through Co-Culture and Column-Based Redox Reactions: Part II
  • +14
  • Dawn Eriksen-Stapleton,
  • Michael King,
  • Guogang Dong,
  • Dhruv Srivastava,
  • Timothy Iskra,
  • Verzhiniya Aho,
  • Kimberly Nguyen,
  • Lia Ingaharro,
  • John J. Scarcelli,
  • Joshua Ochocki,
  • Matthew Gagnon,
  • Robert Hartsough,
  • Hongheng Keo,
  • Courtney Hulme,
  • John Coyne,
  • Cassandra Neubauer,
  • Arch Creasy
Dawn Eriksen-Stapleton
Pfizer Andover

Corresponding Author:[email protected]

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Michael King
Pfizer Andover
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Guogang Dong
Pfizer Andover
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Dhruv Srivastava
Pfizer Andover
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Timothy Iskra
Pfizer Andover
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Verzhiniya Aho
Pfizer Andover
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Kimberly Nguyen
Pfizer Andover
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Lia Ingaharro
Pfizer Andover
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John J. Scarcelli
Pfizer Andover
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Joshua Ochocki
Pfizer Andover
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Matthew Gagnon
Pfizer Andover
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Robert Hartsough
Pfizer Andover
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Hongheng Keo
Pfizer Andover
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Courtney Hulme
Pfizer Andover
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John Coyne
Pfizer Andover
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Cassandra Neubauer
Pfizer Andover
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Arch Creasy
Pfizer Andover
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Abstract

Multispecifics are increasingly being evaluated in the pharmaceutical industry due to the unique mechanisms of action of the molecules, which is enabled by the multiple antigen binding capability. The complexity of these molecules can make production difficult, therefore different approaches for the generation of these molecules have been developed. The approach employed in this study utilizes electrostatic-steering, wherein charge-based differences between two parental homodimer antibodies are used to drive correct heterodimerization during a redox reaction of the partially purified parental homodimers. This strategy results in high conversion to the heterodimer with minimal product-related impurities; however, this method also requires separate bioreactors for each parental homodimer, resulting in complex manufacturing campaigns. This work describes a new bioprocess for electrostatic steering-based multispecifics. This strategy couples two unique components. First, the two separate cell lines are co-cultured, resulting in simultaneous production of both parental homodimers in a single bioreactor. The second component utilizes a column-based redox reaction, wherein the homodimers are captured and the disulfide bonds reduced while bound to the protein A resin using a reductant wash. The column is then eluted and neutralized to allow the reduced parental homodimers to heterodimerize and finally, the addition of an oxidant enables the disulfide bond reformation to complete the formation of the multispecific. This new process is robust and efficient across bench and manufacturing scales, with well-controlled impurity profiles. Through this strategy, the new multispecific bioprocess is more similar to a typical antibody-like bioprocess, enabling more efficient use of clinical and commercial manufacturing resources, while resulting in the production of complex multispecific molecules with minimal product-related impurities.
04 Nov 2024Submitted to Biotechnology and Bioengineering
07 Nov 2024Submission Checks Completed
07 Nov 2024Assigned to Editor
07 Nov 2024Review(s) Completed, Editorial Evaluation Pending
17 Nov 2024Reviewer(s) Assigned