Boronic acid-modified magnetic materials for antibody purification

Vijaykumar L Dhadge, Abid Hussain, Ana M. Azevedo, Raquel Aires-Barros, Ana Cecília Afonso Roque

Research output: Contribution to journalArticle

19 Citations (Scopus)

Abstract

Aminophenyl boronic acids can form reversible covalent ester interactions withcis-diol-containing molecules, serving as a selective tool for binding glycoproteins as antibody molecules that possess oligosaccharides in both the Fv and Fc regions. In this study, amino phenyl boronic acid (APBA) magnetic particles (MPs) were applied for the magnetic separation of antibody molecules. Iron oxide MPs were firstly coated with dextran to avoid non-specific binding and then with 3-glycidyloxypropyl trimethoxysilane to allow further covalent coupling of APBA (APBA_MP). When contacted with pure protein solutions of human IgG (hIgG) and bovine serum albumin (BSA), APBA_MP bound 170 ± 10 mg hIgG g−1MP and eluted 160 ± 5 mg hIgG g−1MP, while binding only 15 ± 5 mg BSA g−1MP. The affinity constant for the interaction between hIgG and APBA_MP was estimated as 4.9 × 105M−1(Ka) with a theoretical maximum capacity of 492 mg hIgG adsorbed g−1MP (Qmax), whereas control particles bound a negligible amount of hIgG and presented an estimated theoretical maximum capacity of 3.1 mg hIgG adsorbed g−1MP (Qmax). APBA_MPs were also tested for antibody purification directly from CHO cell supernatants. The particles were able to bind 98% of IgG loaded and to recover 95% of pure IgG (purity greater than 98%) at extremely mild conditions.
Original languageEnglish
Article number20130875
JournalJournal Of The Royal Society Interface
Volume11
Issue number91
DOIs
Publication statusPublished - 2013

Keywords

  • magnetic particles
  • amino phenyl boronic acid;
  • antibodies
  • glycosylated proteins

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