Comparability of a Three-Dimensional Structure in Biopharmaceuticals Using Spectroscopic Methods
Joint Authors
Villaseñor-Ortega, Francisco
Romero-Díaz, Alexis J.
Pérez, Néstor O.
Medina-Rivero, Emilio
Pérez Medina Martínez, Víctor
Flores-Ortiz, Luis F.
Abad-Javier, Mario E.
Source
Journal of Analytical Methods in Chemistry
Issue
Vol. 2014, Issue 2014 (31 Dec. 2014), pp.1-11, 11 p.
Publisher
Hindawi Publishing Corporation
Publication Date
2014-05-22
Country of Publication
Egypt
No. of Pages
11
Main Subjects
Abstract EN
Protein structure depends on weak interactions and covalent bonds, like disulfide bridges, established according to the environmental conditions.
Here, we present the validation of two spectroscopic methodologies for the measurement of free and unoxidized thiols, as an attribute of structural integrity, using 5,5′-dithionitrobenzoic acid (DTNB) and DyLight Maleimide (DLM) as derivatizing agents.
These methods were used to compare Rituximab and Etanercept products from different manufacturers.
Physicochemical comparability was demonstrated for Rituximab products as DTNB showed no statistical differences under native, denaturing, and denaturing-reducing conditions, with Student’s t-test P values of 0.6233, 0.4022, and 0.1475, respectively.
While for Etanercept products no statistical differences were observed under native (P=0.0758) and denaturing conditions (P=0.2450), denaturing-reducing conditions revealed cysteine contents of 98% and 101%, towards the theoretical value of 58, for the evaluated products from different Etanercept manufacturers.
DLM supported equality between Rituximab products under native (P=0.7499) and denaturing conditions (P=0.8027), but showed statistical differences among Etanercept products under native conditions (P<0.001).
DLM suggested that Infinitam has fewer exposed thiols than Enbrel, although DTNB method, circular dichroism (CD), fluorescence (TCSPC), and activity (TNFα neutralization) showed no differences.
Overall, this data revealed the capabilities and drawbacks of each thiol quantification technique and their correlation with protein structure.
American Psychological Association (APA)
Pérez Medina Martínez, Víctor& Abad-Javier, Mario E.& Romero-Díaz, Alexis J.& Villaseñor-Ortega, Francisco& Pérez, Néstor O.& Flores-Ortiz, Luis F.…[et al.]. 2014. Comparability of a Three-Dimensional Structure in Biopharmaceuticals Using Spectroscopic Methods. Journal of Analytical Methods in Chemistry،Vol. 2014, no. 2014, pp.1-11.
https://search.emarefa.net/detail/BIM-510759
Modern Language Association (MLA)
Pérez Medina Martínez, Víctor…[et al.]. Comparability of a Three-Dimensional Structure in Biopharmaceuticals Using Spectroscopic Methods. Journal of Analytical Methods in Chemistry No. 2014 (2014), pp.1-11.
https://search.emarefa.net/detail/BIM-510759
American Medical Association (AMA)
Pérez Medina Martínez, Víctor& Abad-Javier, Mario E.& Romero-Díaz, Alexis J.& Villaseñor-Ortega, Francisco& Pérez, Néstor O.& Flores-Ortiz, Luis F.…[et al.]. Comparability of a Three-Dimensional Structure in Biopharmaceuticals Using Spectroscopic Methods. Journal of Analytical Methods in Chemistry. 2014. Vol. 2014, no. 2014, pp.1-11.
https://search.emarefa.net/detail/BIM-510759
Data Type
Journal Articles
Language
English
Notes
Includes bibliographical references
Record ID
BIM-510759