Biomedical Engineering Reference
In-Depth Information
addition to unexpected staining patterns from variations in cell growth and protein
expression profiles. Assay validation parameters follow those detailed in the literature
and referenced here, using the flow cytometer with appropriate, applicable, and well-
characterized reagents. Comprehending and mitigating the unique technical chal-
lenges, defining assay readout, and understanding internal variability of the assay
combined with applying an assay cut point that defines a sample as positive or
negative allow the bioanalytical lab to use flow cytometry as a powerful tool in
immunogenicity testing.
ACKNOWLEDGMENTS
The authors would like to thank Terri Caiazzo, Alison Joyce, Keirsten Seiger, and Lisa
Cherry for their technical assistance and John Nowak, JimMcNally, Corinna Fiorotti,
Terri Caiazzo, Alison Joyce, and Yuanxin Xu for their review of this chapter. In
particular, we appreciate the invaluable suggestions received from Bonita Rup and
Susan Richards.
REFERENCES
1. Gupta S et al. Recommendations for the design, optimization, and qualification of cell-
based assays used for the detection of neutralizing antibody responses elicited to biological
therapeutics. J. Immunol. Methods 2007;321(1-2):1-18.
2. Ponce R et al. Immunogenicity of biologically-derived therapeutics: assessment and
interpretation of nonclinical safety studies. Regul. Toxicol. Pharmacol. 2009;54
(2):164-182.
3. Krutzik PO, Nolan GP. Intracellular phospho-protein staining techniques for flow
cytometry: monitoring single cell signaling events. Cytometry A 2003;55(2):61-70.
4. Rothaeusler K, Baumgarth N. Evaluation of intranuclear BrdU detection procedures for
use in multicolor flow cytometry. Cytometry A 2006;69(4):249-259.
5. McCoy JP, Jr. Basic principles of flow cytometry. Hematol. Oncol. Clin. North Am.
2002;16(2):229-243.
6. Rup B, O'Hara D. Critical ligand binding reagent preparation/selection: when specificity
depends on reagents. AAPS J. 2007;9(2):E148-55.
7. Holden TM, Tom F, Laurel EN, Trotter, J. Selecting fluorochrome conjugates for
maximum sensitivity. Cytometry A 2004;62A:169-173.
8. Nomura L, V. C. Maino, and H. T. Maecker. Standardization and optimization of
multiparameter intracellular cytokine staining. Cytometry A 2008.73(11):984-991.
9. Herzenberg L. A et al. Interpreting flow cytometry data: a guide for the perplexed. Nat.
Immunol. 2006;7(7):681-685.
10. Tung JWet al. New approaches to fluorescence compensation and visualization of FACS
data. Clin. Immunol. 2004.110(3):277-83.
11. Ritter N, Wiebe M. Validating critical reagents used in CGMP analytical testing: ensuring
method integrity and reliable assay performance. BioPharm. 2001;14(5):12-21.
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