Critical evaluation and guidance for using the Coulter method for counting subvisible particles in protein solutions.

In this paper, we evaluate the suitability of Coulter method for detecting and quantifying subvisible particles in protein solutions and compare results with other particle-counting technologies. The effects of key instrument and operational parameters such as aperture diameter, solution conductivity, and cleaning procedures are demonstrated. Degraded and nondegraded intravenous immunoglobulin and human serum albumin were chosen as model proteins and sample types for this evaluation. Multisizer™4 was able to obtain reproducible and linear particle counts; however, customized analysis and cleaning procedures are needed depending on the protein analyzed and the sample type (degraded or nondegraded). The Coulter method consistently detected more particles than micro-flow imaging and light obscuration. The presence of translucent particles likely accounts for this observation because detection by the Coulter method does not depend on the optical properties of the particles or solution.

[1]  J. Carletta,et al.  Capacitive Coulter counting: detection of metal wear particles in lubricant using a microfluidic device , 2009 .

[2]  Theodore W Randolph,et al.  Subvisible particle counting provides a sensitive method of detecting and quantifying aggregation of monoclonal antibody caused by freeze-thawing: insights into the roles of particles in the protein aggregation pathway. , 2011, Journal of pharmaceutical sciences.

[3]  D C Carter,et al.  Three-dimensional structure of human serum albumin. , 1989, Science.

[4]  T. Habermann,et al.  Evaluation of the Coulter Counter model S-Plus IV. , 1985, American journal of clinical pathology.

[5]  S. Singh,et al.  An industry perspective on the monitoring of subvisible particles as a quality attribute for protein therapeutics. , 2010, Journal of pharmaceutical sciences.

[6]  D. Volkin,et al.  Development of a microflow digital imaging assay to characterize protein particulates during storage of a high concentration IgG1 monoclonal antibody formulation. , 2010, Journal of pharmaceutical sciences.

[7]  R. Xu,et al.  Comparison of sizing small particles using different technologies , 2003 .

[8]  S. Ben‐Sasson,et al.  Electrical sizing of particles in suspensions. I. Theory. , 1969, Biophysical journal.

[9]  Theodore W Randolph,et al.  Aggregation of a monoclonal antibody induced by adsorption to stainless steel , 2010, Biotechnology and bioengineering.

[10]  Hanns-Christian Mahler,et al.  Protein aggregation: pathways, induction factors and analysis. , 2009, Journal of pharmaceutical sciences.

[11]  Marshall D Graham,et al.  The Coulter Principle: Foundation of an Industry , 2003 .

[12]  Huub Schellekens,et al.  Antibody response to aggregated human interferon alpha2b in wild-type and transgenic immune tolerant mice depends on type and level of aggregation. , 2006, Journal of Pharmacy and Science.

[13]  J. Mulder,et al.  Heterogeneity of monoclonal immunoglobulin-G proteins studied by isoelectric focusing. , 1974, Clinica chimica acta; international journal of clinical chemistry.

[14]  Chi-Ting Huang,et al.  Quantitation of protein particles in parenteral solutions using micro-flow imaging. , 2009, Journal of pharmaceutical sciences.

[15]  S. Ben‐Sasson,et al.  Electrical sizing of particles in suspensions. 3. Rigid spheroids and red blood cells. , 1972, Biophysical journal.

[16]  A. Rosenberg,et al.  Effects of protein aggregates: An immunologic perspective , 2006, The AAPS Journal.

[17]  U. Zimmermann,et al.  Particles in a homogeneous electrical field: A model for the electrical breakdown of living cells in a coulter counter , 1979 .

[18]  S. Ben‐Sasson,et al.  Electrical sizing of particles in suspensions. II. Experiments with rigid spheres. , 1969, Biophysical journal.

[19]  Steven Kozlowski,et al.  Overlooking subvisible particles in therapeutic protein products: gaps that may compromise product quality. , 2009, Journal of pharmaceutical sciences.

[20]  S. Shire,et al.  Characterization of Particles in Protein Solutions: Reaching the Limits of Current Technologies , 2010, The AAPS Journal.

[21]  P H Pinkerton,et al.  An assessment of the Coulter counter model S , 1970, Journal of clinical pathology.

[22]  T. Lindmo,et al.  Flow cytometry and sorting , 1979 .

[23]  J. Johnson.  Thermal Agitation of Electricity in Conductors , 1928 .

[24]  Jared S. Bee,et al.  Monoclonal antibody interactions with micro- and nanoparticles: adsorption, aggregation, and accelerated stress studies. , 2009, Journal of pharmaceutical sciences.

[25]  Amber Haynes Fradkin,et al.  Immunogenicity of aggregates of recombinant human growth hormone in mouse models. , 2009, Journal of pharmaceutical sciences.

[26]  Theodore W Randolph,et al.  IgG particle formation during filling pump operation: a case study of heterogeneous nucleation on stainless steel nanoparticles. , 2009, Journal of pharmaceutical sciences.

[27]  Wei Wang,et al.  Antibody structure, instability, and formulation. , 2007, Journal of pharmaceutical sciences.

[28]  R. Dudley.  The particle size analysis of soils and its use in forensic science--the determination of particle size distributions within the silt and sand fractions. , 1976, Journal - Forensic Science Society.

[29]  H. K. Wilson.  Measurements and Counts of Large Protein Particles in Sterile Concentrated Skimmilk , 1973 .

[30]  G. V. D. Plaats,et al.  A study on the sizing process of an instrument based on the electrical sensing zone principle. Part 2. The influence of particle porosity , 1983 .

[31]  L. Narhi,et al.  A critical review of analytical methods for subvisible and visible particles. , 2009, Current pharmaceutical biotechnology.

[32]  D. K. Sharma,et al.  Micro-Flow Imaging: Flow Microscopy Applied to Sub-visible Particulate Analysis in Protein Formulations , 2010, The AAPS Journal.

[33]  B. Bain.  An assessment of the three-population differential count on the Coulter Counter Model S Plus IV. , 1986, Clinical and laboratory haematology.