Theoretical approximations and experimental extinction coefficients of biopharmaceuticals

详细信息    查看全文

• 作者：Mariana P. Miranda-Hernández…
• 关键词：Refractive index increment ; Extinction coefficient ; Protein quantitation ; Biopharmaceuticals ; Dry weight
• 刊名：Analytical and Bioanalytical Chemistry
• 出版年：2016
• 出版时间：February 2016
• 年：2016
• 卷：408
• 期：5
• 页码：1523-1530
• 全文大小：597 KB
• 参考文献：1.Bradford MM. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976;72:248–54.CrossRef
  2.Lowry OH, Rosebrough NJ, Farr AL, Randall RJ. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951;193:265–75.
  3.Layne E. Spectrophotometric and turbidimetric methods for measuring proteins. Methods Enzymol. 1957;3:447–55.CrossRef
  4.Scopes RK. Measurement of protein by spectrophotometry at 205 nm. Anal Biochem. 1974;59:277–82.CrossRef
  5.Gorball AG, Bardawill CJ, David MM. Determination of serum proteins by means of the biuret reaction. J Biol Chem. 1949;177:751–66.
  6.Gill SC, von Hippel PH. Calculation of protein extinction coefficients from amino acid sequence data. Anal Biochem. 1989;182:319–26.CrossRef
  7.Pace CN, Vajdos F, Gray T. How to measure and predict the molar absorption coefficient of a protein. Protein Sci. 1995;4:2411–23.CrossRef
  8.Noble JE, Knight AE, Reason AJ, Di Matola A, Bailey MJ. A comparison of protein quantitation assays for biopharmaceutical applications. Mol Biotechnol. 2007;37:99–111.CrossRef
  9.Stoscheck CM. Quantitation of protein. Methods Enzymol. 1990;182:50–68.CrossRef
  10.ICH (1999) International Conference on Harmonisation of Technical Requirements for Registration of Pharmaceuticals for Human Use. ICH Q6B: Specifications: test procedures and acceptance criteria for biotechnological/biological products.
  11.Espinosa-de la-Garza CE, Miranda-Hernández MP, Acosta-Flores L, Pérez NO, Flores-Ortiz LF, Medina-Rivero E. Analysis of therapeutic proteins and peptides using multiangle light scattering coupled to ultra high performance liquid chromatography. J Sep Sci. 2015;38:1537–43.CrossRef
  12.Podzimek S. Light scattering, size exclusion chromatography and asymmetric flow field fractionation. New Jersey: Wiley; 2011.CrossRef
  13.Flores-Ortiz LF, Campos-García VR, Perdomo-Abúndez FC, Pérez NO, Medina-Rivero E. Physicochemical properties of rituximab. J Liq Chromatogr Relat Technol. 2014;37:1438–52.CrossRef
  14.Edelhoch H. Spectroscopic determination of tryptophan and tyrosine in proteins. Biochemistry. 1967;6:1948–54.CrossRef
  15.Artimo P, Jonnalagedda M, Arnold K, Baatin D, Csardi G, de Castro E, et al. ExPASy: SIB bioinformatics resource portal. Nucleic Acids Res. 2012;40(W1):W597–603.CrossRef
  16.Nozaki Y. Determination of the concentration of protein by dry-weight, a comparison with spectroscopic methods. Arch Biochem Biophys. 1986;249:437–46.CrossRef
  17.Kupke DW, Dorrier TE. Protein concentration measurements: the dry weight. Methods Enzymol. 1978;48:155–62.CrossRef
  18.McMeekin TL, Wilensky M, Groves ML. Refractive indices of proteins in relation to amino acid composition and specific volume. Biochem Biophys. 1962;7:151–6.CrossRef
  19.Zhao H, Brown PH, Schuck P. On the distribution of protein refractive index increments. Biophys J. 2011;100:2309–17.CrossRef
  20.Schuck P. Size distribution analysis of macromolecules by sedimentation velocity ultracentrifugation and Lamm equation modeling. Biophys J. 2000;78:1606–19.CrossRef
  21.Wen J, Arakawa T. Refractive index of proteins in aqueous sodium chloride. Anal Biochem. 2000;280:327–9.CrossRef
  22.Ye H. Simultaneous determination of protein aggregation, degradation, and absolute molecular weight by size exclusion chromatography-multiangle light scattering. Anal Biochem. 2006;356:76–85.CrossRef
  23.Fedosov SN, Fedosova NU, Berglund L, Moestrup SK, Nexo E, Petersen TE. Assembly of the intrinsic factor domains and oligomerization of the protein in the presence of cobalamin. Biochemistry. 2004;43:15095–102.CrossRef
  24.Mahendiran K, Elie C, Nebel JC, Ryan A, Pierscionek BK. Primary sequence contribution to the optical function of the eye lens. Sci Rep. 2014;4:5195.CrossRef
  25.Crisman RL, Randolph TW. Refolding of proteins from inclusion bodies is favored by a diminished hydrophobic effect at elevated pressures. Biotechnol Bioeng. 2008;102:483–92.CrossRef
  
• 作者单位：Mariana P. Miranda-Hernández (1)
  Elba R. Valle-González (1)
  David Ferreira-Gómez (1)
  Néstor O. Pérez (1)
  Luis F. Flores-Ortiz (1)
  Emilio Medina-Rivero (1)
  
  1. Unidad de Investigación y Desarrollo, Probiomed S.A. de C.V., Cruce de carreteras Acatzingo-Zumpahuacán, 52400, Tenancingo, Mexico
  
• 刊物类别：Chemistry and Materials Science
• 刊物主题：Chemistry
  Analytical Chemistry
  Food Science
  Inorganic Chemistry
  Physical Chemistry
  Monitoring, Environmental Analysis and Environmental Ecotoxicology
  
• 出版者：Springer Berlin / Heidelberg
• ISSN：1618-2650

文摘

UV spectrophotometric measurement is a widely accepted and standardized routine analysis for quantitation of highly purified proteins; however, the reliability of the results strictly depends on the accuracy of the employed extinction coefficients. In this work, an experimental estimation of the differential refractive index (dn/dc), based on dry weight measurements, was performed in order to determine accurate extinction coefficients for four biotherapeutic proteins and one synthetic copolymer after separation in a size-exclusion ultra-performance liquid chromatograph coupled to an ultraviolet, multiangle light scattering and refractive index (SE-UPLC-UV-MALS-RI) multidetection system. The results showed small deviations with respect to theoretical values, calculated from the specific amino acid sequences, for all the studied immunoglobulins. Nevertheless, for proteins like etanercept and glatiramer acetate, several considerations, such as glycan content, partial specific volume, polarizability, and higher order structure, should be considered to properly calculate theoretical extinction coefficient values. Herein, these values were assessed with simple approximations. The precision of the experimentally obtained extinction coefficients, and its convergence towards the theoretical values, makes them useful for characterization and comparability exercises. Also, these values provide insight into the absorbance and scattering properties of the evaluated proteins. Overall, this methodology is capable of providing accurate extinction coefficients useful for development studies.