A critical review of analytical methods for subvisible and visible particles

Curr Pharm Biotechnol. 2009 Jun;10(4):373-81. doi: 10.2174/138920109788488905.

Abstract

The subvisible and visible particles present in a solution are often classified based on size, and are quantified by the actual number of particles present rather than by weight or molar amounts. The analysis of these particles in protein therapeutics are governed by compendial methods and the regulatory agencies, and the methods available to measure them originally evolved focusing on potential safety issues, including capillary occlusion and immunogenicity, that might arise from their presence. Ultracentrifugation, size exclusion chromatography, etc., discussed in previous articles, can be used to analyze aggregates of less than 0.10 microns. This article will focus on methods for analyzing and quantitating sub visible particles (SbVP) of 2 microns or larger. At the present time there is no routine method for quantitating sub visible particles (SbVP) between 0.1 microns and 2 microns. The most common technique for quantitating the amount of subvisible particles between 2 and 100 microns is the light obscuration method. This technique can determine size and amount of particles, but cannot differentiate between the types of particles, such as protein particles, foreign material, micro bubbles or silicone oil droplets, that can be present in protein solutions. The difficulties in adapting this method, originally developed for small molecule drugs for IV administration, to protein therapeutics delivered subcutaneously is discussed. The flow imaging techniques can determine morphology and optical characteristics of the particles, but still not identify the chemical composition. Other methods that can also be used, but are applicable for characterization purposes only, are discussed. The primary method for quantitating visible particles is visual inspection, a method that can be subjective and relies on adequate training of the human inspectors. Automated methods for visible particle determination are being developed. Identification of the chemical composition of isolated particles greater than about 50 microns is possible using several micro-spectroscopic methods, and these will also be discussed.

Publication types

  • Review

MeSH terms

  • Chemical Fractionation / methods*
  • Microspheres
  • Multiprotein Complexes / chemistry
  • Multiprotein Complexes / isolation & purification
  • Nephelometry and Turbidimetry / methods*
  • Particle Size
  • Proteins / chemistry*
  • Proteins / isolation & purification*
  • Spectrum Analysis / methods*
  • Technology, Pharmaceutical / methods*

Substances

  • Multiprotein Complexes
  • Proteins