PMID- 34224732
OWN - NLM
STAT- MEDLINE
DCOM- 20211028
LR  - 20211028
IS  - 1520-6017 (Electronic)
IS  - 0022-3549 (Linking)
VI  - 110
IP  - 11
DP  - 2021 Nov
TI  - Identification and Characterization of Polysorbate-Degrading Enzymes in a 
      Monoclonal Antibody Formulation.
PG  - 3558-3567
LID - S0022-3549(21)00353-1 [pii]
LID - 10.1016/j.xphs.2021.06.033 [doi]
AB  - Degradation of polysorbate (PS) by hydrolytically active host cell proteins 
      (HCPs) in drug products may impair the protein-stabilizing properties of PS and 
      lead to the formation of particles due to the accumulation of poorly soluble free 
      fatty acids upon long-term storage. The identification of the causative enzymes 
      is challenging due to their low-abundance even when using state-of-the-art 
      instrumentation and workflows. To overcome these challenges, we developed a 
      rigorous enrichment strategy for HCPs, utilizing both Protein A and anti-HCP 
      affinity chromatography, which facilitated the in-depth characterization of the 
      HCP population in a monoclonal antibody formulation prone to PS hydrolysis. Based 
      on the HCPs identified by liquid chromatography coupled to tandem mass 
      spectrometry, a number of enzymes annotated as hydrolases were recombinantly 
      expressed and characterized in terms of polysorbate degradation. Among the 
      selected candidates, Lipoprotein Lipase, Lysosomal Acid Lipase (LIPA) and 
      Palmitoyl-Protein Thioesterase 1 (PPT1) exhibited notable activity towards PS. To 
      our knowledge, this is the first report to identify LIPA and PPT1 as residual 
      HCPs that can contribute to PS degradation in a biological product.
CI  - Copyright (c) 2021 American Pharmacists Association. All rights reserved.
FAU - Graf, Tobias
AU  - Graf T
AD  - Roche Diagnostics GmbH, Nonnenwald 2, 82377 Penzberg, Germany. Electronic 
      address: Tobias.Graf@roche.com.
FAU - Tomlinson, Anthony
AU  - Tomlinson A
AD  - Pharma Technical Development, Genentech, 1 DNA Way, South San Francisco, 
      California, USA.
FAU - Yuk, Inn H
AU  - Yuk IH
AD  - Pharma Technical Development, Genentech, 1 DNA Way, South San Francisco, 
      California, USA.
FAU - Kufer, Regina
AU  - Kufer R
AD  - Roche Diagnostics GmbH, Nonnenwald 2, 82377 Penzberg, Germany.
FAU - Spensberger, Bernhard
AU  - Spensberger B
AD  - Roche Diagnostics GmbH, Nonnenwald 2, 82377 Penzberg, Germany.
FAU - Falkenstein, Roberto
AU  - Falkenstein R
AD  - Roche Diagnostics GmbH, Nonnenwald 2, 82377 Penzberg, Germany.
FAU - Shen, Amy
AU  - Shen A
AD  - Pharma Technical Development, Genentech, 1 DNA Way, South San Francisco, 
      California, USA.
FAU - Li, Hong
AU  - Li H
AD  - Pharma Technical Development, Genentech, 1 DNA Way, South San Francisco, 
      California, USA.
FAU - Duan, Dana
AU  - Duan D
AD  - Pharma Technical Development, Genentech, 1 DNA Way, South San Francisco, 
      California, USA.
FAU - Liu, Wenqiang
AU  - Liu W
AD  - Pharma Technical Development, Genentech, 1 DNA Way, South San Francisco, 
      California, USA.
FAU - Wohlrab, Stefanie
AU  - Wohlrab S
AD  - Roche Diagnostics GmbH, Nonnenwald 2, 82377 Penzberg, Germany.
FAU - Edelmann, Franziska
AU  - Edelmann F
AD  - Roche Diagnostics GmbH, Nonnenwald 2, 82377 Penzberg, Germany.
FAU - Leiss, Michael
AU  - Leiss M
AD  - Roche Diagnostics GmbH, Nonnenwald 2, 82377 Penzberg, Germany.
LA  - eng
PT  - Journal Article
DEP - 20210702
PL  - United States
TA  - J Pharm Sci
JT  - Journal of pharmaceutical sciences
JID - 2985195R
RN  - 0 (Antibodies, Monoclonal)
RN  - 0 (Polysorbates)
SB  - IM
MH  - *Antibodies, Monoclonal
MH  - Chromatography, Liquid
MH  - Hydrolysis
MH  - *Polysorbates
MH  - Tandem Mass Spectrometry
OTO - NOTNLM
OT  - Antibody drug(s)
OT  - Enzyme(s)
OT  - Hydrolysis
OT  - Liquid chromatography-mass spectrometry (LC-MS)
OT  - Monoclonal antibody(s)
OT  - Protein formulation(s)
OT  - Surfactant(s)
COIS- Declaration of Competing Interests The authors declare that they have no known 
      competing financial interests or personal relationships that could have appeared 
      to influence the work reported in this paper.
EDAT- 2021/07/06 06:00
MHDA- 2021/10/29 06:00
CRDT- 2021/07/05 20:12
PHST- 2021/05/21 00:00 [received]
PHST- 2021/06/28 00:00 [revised]
PHST- 2021/06/29 00:00 [accepted]
PHST- 2021/07/06 06:00 [pubmed]
PHST- 2021/10/29 06:00 [medline]
PHST- 2021/07/05 20:12 [entrez]
AID - S0022-3549(21)00353-1 [pii]
AID - 10.1016/j.xphs.2021.06.033 [doi]
PST - ppublish
SO  - J Pharm Sci. 2021 Nov;110(11):3558-3567. doi: 10.1016/j.xphs.2021.06.033. Epub 
      2021 Jul 2.