PMID- 33158205
OWN - NLM
STAT- PubMed-not-MEDLINE
LR  - 20201128
IS  - 2076-2607 (Print)
IS  - 2076-2607 (Electronic)
IS  - 2076-2607 (Linking)
VI  - 8
IP  - 11
DP  - 2020 Nov 4
TI  - Bacterial Endotoxin Testing-Fast Endotoxin Masking Kinetics in the Presence of 
      Lauryldimethylamine Oxide.
LID - 10.3390/microorganisms8111728 [doi]
LID - 1728
AB  - For release of parenteral drug products, bacterial endotoxin testing is one of a 
      panel of necessary tests. In order to ensure the validity of such tests, various 
      controls are performed, including demonstration of compendial method suitability 
      or method qualification. In addition to compendial suitability testing, quality 
      control (QC) sample hold-time studies are requested by authorities like the Food 
      and Drug Administration (FDA) as described in "Guidance for Industry: Pyrogen and 
      Endotoxins Testing." It is requested to be determine whether the ability to 
      detect endotoxins can be affected by storage and handling of the sample to be 
      tested. To accomplish these studies, endotoxin is introduced or spiked into the 
      undiluted product and held for a certain period of time in process-representative 
      containers. This time period reflects procedural maximum QC sample hold time from 
      sampling until analysis. Inadequate detection of endotoxin can be caused by 
      adsorption of endotoxin to container surfaces or molecular masking effects, in 
      which the binding sites on the endotoxin molecules are prevented from triggering 
      the enzymatic cascade necessary in the assay, are obscured. The endotoxin may 
      form macromolecular structures, such as sheets or blebs, or the binding sites may 
      otherwise be rendered unavailable due to the sample matrix composition. In either 
      case, the endotoxin assay may yield falsely low results if and when masking 
      occurs. In this work, the QC sample hold times of different in-process controls 
      within the production process of a biopharmaceutical product were analyzed. One 
      out of eight different samples showed a strong masking of endotoxin. Analysis of 
      the sample composition revealed that either kifunensine, mycophenolic acid (MPA), 
      or lauryl-N, N-dimethylamine oxide (LDAO) was responsible for masking. Further 
      analysis clearly identified LDAO as the root cause for masking. A novel one-step 
      mechanism for LDAO-induced endotoxin masking is proposed. The principle is 
      similar to an already-proposed two-step mechanism for endotoxin masking, but the 
      LDAO case combines these two steps: the disturbance of the salt bridges and 
      hydrophobic interactions with LPS in one molecule. These molecular interactions 
      occur quickly when both endotoxin and LDAO are present in the same matrix. Thus, 
      depending on the masking agents, low endotoxin recovery (LER) can occur 
      regardless of the QC sample hold duration.
FAU - Bech Orving, Rene
AU  - Bech Orving R
AD  - FUJIFILM Diosynth Biotechnologies, 3400 Hillerod, Denmark.
FAU - Carpenter, Bill
AU  - Carpenter B
AD  - Biogen, Cambridge, MA 02142, USA.
FAU - Roth, Steffen
AU  - Roth S
AD  - Microcoat Biotechnologie GmbH, 82347 Bernried, Germany.
FAU - Reich, Johannes
AU  - Reich J
AUID- ORCID: 0000-0002-1200-870X
AD  - Microcoat Biotechnologie GmbH, 82347 Bernried, Germany.
FAU - Kallipolitis, Birgitte Haahr
AU  - Kallipolitis B
AD  - Department of Biochemistry and Molecular Biology, University of Southern Denmark, 
      5230 Odense, Denmark.
FAU - Sonne-Hansen, Jacob
AU  - Sonne-Hansen J
AD  - FUJIFILM Diosynth Biotechnologies, 3400 Hillerod, Denmark.
LA  - eng
PT  - Journal Article
DEP - 20201104
PL  - Switzerland
TA  - Microorganisms
JT  - Microorganisms
JID - 101625893
PMC - PMC7694283
OTO - NOTNLM
OT  - LAL
OT  - LDAO
OT  - LER
OT  - LPS
OT  - endotoxin
OT  - lauryldimethylamine oxide
OT  - limulus amoebocyte lysate
OT  - low endotoxin recovery
OT  - masking
COIS- The authors declare no conflict of interest.
EDAT- 2020/11/08 06:00
MHDA- 2020/11/08 06:01
PMCR- 2020/11/04
CRDT- 2020/11/07 01:03
PHST- 2020/10/09 00:00 [received]
PHST- 2020/10/30 00:00 [revised]
PHST- 2020/10/30 00:00 [accepted]
PHST- 2020/11/07 01:03 [entrez]
PHST- 2020/11/08 06:00 [pubmed]
PHST- 2020/11/08 06:01 [medline]
PHST- 2020/11/04 00:00 [pmc-release]
AID - microorganisms8111728 [pii]
AID - microorganisms-08-01728 [pii]
AID - 10.3390/microorganisms8111728 [doi]
PST - epublish
SO  - Microorganisms. 2020 Nov 4;8(11):1728. doi: 10.3390/microorganisms8111728.