Automated capillary Western dot blot method for the identity of a 15-valent pneumococcal conjugate vaccine

Anal Biochem. 2015 Jun 1:478:33-9. doi: 10.1016/j.ab.2015.03.021. Epub 2015 Mar 23.

Abstract

Simple Western is a new technology that allows for the separation, blotting, and detection of proteins similar to a traditional Western except in a capillary format. Traditionally, identity assays for biological products are performed using either an enzyme-linked immunosorbent assay (ELISA) or a manual dot blot Western. Both techniques are usually very tedious, labor-intensive, and complicated for multivalent vaccines, and they can be difficult to transfer to other laboratories. An advantage this capillary Western technique has over the traditional manual dot blot Western method is the speed and the automation of electrophoresis separation, blotting, and detection steps performed in 96 capillaries. This article describes details of the development of an automated identity assay for a 15-valent pneumococcal conjugate vaccine, PCV15-CRM197, using capillary Western technology.

Keywords: CRM197; Capillary Western; Dot blot; Pneumococcal conjugate vaccine; Polysaccharide; Simple Western.

MeSH terms

  • Animals
  • Bacterial Proteins / chemistry
  • Blotting, Western / methods*
  • Enzyme-Linked Immunosorbent Assay
  • Humans
  • Pneumococcal Infections / prevention & control
  • Pneumococcal Vaccines / chemistry*
  • Vaccines, Conjugate / chemistry*

Substances

  • Bacterial Proteins
  • Pneumococcal Vaccines
  • Vaccines, Conjugate
  • CRM197 (non-toxic variant of diphtheria toxin)