Purification of antibody against Ara h 2 by a homemade immunoaffinity chromatography column

Prep Biochem Biotechnol. 2017 Sep 14;47(8):841-846. doi: 10.1080/10826068.2017.1342265. Epub 2017 Jun 21.

Abstract

Antibodies are used extensively in numerous applications both in vivo and in vitro. To purify anti-Ara h 2 polyclonal antibody, a homemade immunoaffinity chromatography (IAC) column method was established. The properties of homemade column were compared with those of the mAb affinity protein G (MPG) agarose high flow, a commercially available column successfully used in capturing polyclonal antibodies. During antibody purification from rabbits' antiserum against Ara h 2, the column capacity, recovery, and purification factor were characterized for IAC and MPG. The homemade IAC could separate the corresponding antibody with higher specificity and lower cost but with lower recovery and column capacity than those of MPG. Thus, the homemade IAC is a specific, inexpensive, and suitable method that can be used for various laboratory purifications.

Keywords: Ara h 2; IAC; MPG; PcAb; immunoaffinity; purification.

MeSH terms

  • 2S Albumins, Plant / immunology*
  • Animals
  • Antibodies / immunology*
  • Antibodies / isolation & purification*
  • Antigens, Plant / immunology*
  • Arachis / immunology*
  • Chromatography, Affinity / methods*
  • Glycoproteins / immunology*
  • Rabbits

Substances

  • 2S Albumins, Plant
  • Antibodies
  • Antigens, Plant
  • Ara h 2 allergen, Arachis hypogaea
  • Glycoproteins