PMID- 22119514
OWN - NLM
STAT- MEDLINE
DCOM- 20120419
LR  - 20191210
IS  - 1872-7905 (Electronic)
IS  - 0022-1759 (Linking)
VI  - 376
IP  - 1-2
DP  - 2012 Feb 28
TI  - Implementation of design of experiments (DOE) in the development and validation 
      of a cell-based bioassay for the detection of anti-drug neutralizing antibodies 
      in human serum.
PG  - 32-45
LID - 10.1016/j.jim.2011.11.004 [doi]
AB  - The administration of biological therapeutics can potentially elicit the 
      development of neutralizing antibodies (NAbs) to the therapeutic drug in 
      patients, which could have a significant impact on drug efficacy and safety. A 
      rigorous in vitro cell-based assay for the detection of NAbs is critical for the 
      assessment of the immunogenicity profile of the therapeutic drug. Conatumumab is 
      a fully human monoclonal agonist antibody directed against the extracellular 
      domain of human TRAIL receptor 2 (TR-2). It is being investigated as a cancer 
      treatment because it is able to induce apoptosis in sensitive tumor cells. This 
      report demonstrates how statistically designed experiments could be employed 
      effectively in different stages of a NAb bioassay life cycle in order to 
      characterize, optimize and stabilize the assay with added benefit of resource 
      efficiency. By combining the approach of design of experiments (DOE) with subject 
      matter expertise and experience, we were able to understand thoroughly how assay 
      parameters affect the performance of the assay individually and interactively, 
      identify the key assay parameters, define assay operating ranges and finally 
      achieve a robust and sensitive cell-based assay for the detection of NAbs to 
      Conatumumab. With the goal of developing a cell-based bioassay that is highly 
      optimized for sensitivity, specificity, precision, and robustness, we performed 2 
      DOE experiments for assay optimization and 1 DOE experiment to validate assay 
      robustness. We evaluated key operating parameters of the assay such as cell 
      number, percentage of serum matrix, concentration of the therapeutic drug, 
      concentration of the cross-linker, length of various incubation steps, cell age, 
      interval between cell subculture and bioassay time, and detection equipment.
CI  - Copyright (c) 2011 Elsevier B.V. All rights reserved.
FAU - Chen, Xinyi C
AU  - Chen XC
AD  - Clinical Immunology Department, Amgen Inc., One Amgen Center Drive, Thousand 
      Oaks, CA 91320, USA.
FAU - Zhou, Lei
AU  - Zhou L
FAU - Gupta, Shalini
AU  - Gupta S
FAU - Civoli, Francesca
AU  - Civoli F
LA  - eng
PT  - Journal Article
PT  - Validation Study
DEP - 20111112
PL  - Netherlands
TA  - J Immunol Methods
JT  - Journal of immunological methods
JID - 1305440
RN  - 0 (Antibodies, Monoclonal)
RN  - 0 (Antibodies, Neutralizing)
RN  - 1P48L61KM0 (conatumumab)
RN  - EC 3.4.22.- (Caspases)
SB  - IM
MH  - Antibodies, Monoclonal/*immunology
MH  - Antibodies, Neutralizing/blood/*immunology
MH  - Biological Assay/*methods
MH  - Caspases/blood/*immunology
MH  - Cell Line, Tumor
MH  - Humans
MH  - Limit of Detection
MH  - Models, Statistical
MH  - Sensitivity and Specificity
EDAT- 2011/11/29 06:00
MHDA- 2012/04/20 06:00
CRDT- 2011/11/29 06:00
PHST- 2010/12/04 00:00 [received]
PHST- 2011/11/01 00:00 [revised]
PHST- 2011/11/07 00:00 [accepted]
PHST- 2011/11/29 06:00 [entrez]
PHST- 2011/11/29 06:00 [pubmed]
PHST- 2012/04/20 06:00 [medline]
AID - S0022-1759(11)00310-3 [pii]
AID - 10.1016/j.jim.2011.11.004 [doi]
PST - ppublish
SO  - J Immunol Methods. 2012 Feb 28;376(1-2):32-45. doi: 10.1016/j.jim.2011.11.004. 
      Epub 2011 Nov 12.