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Crivianu-Gaita V, Romaschin A, Thompson M. High efficiency reduction capability for the formation of Fab׳ antibody fragments from F(ab). Biochem Biophys Rep. 2015;2:23-28doi: 10.1016/j.bbrep.2015.04.004.
Crivianu-Gaita, V., Romaschin, A., & Thompson, M. (2015). High efficiency reduction capability for the formation of Fab׳ antibody fragments from F(ab). Biochemistry and biophysics reports, 223-28. https://doi.org/10.1016/j.bbrep.2015.04.004
Crivianu-Gaita, Victor, et al. "High efficiency reduction capability for the formation of Fab׳ antibody fragments from F(ab)." Biochemistry and biophysics reports vol. 2 (2015): 23-28. doi: https://doi.org/10.1016/j.bbrep.2015.04.004
Crivianu-Gaita V, Romaschin A, Thompson M. High efficiency reduction capability for the formation of Fab׳ antibody fragments from F(ab). Biochem Biophys Rep. 2015 Apr 25;2:23-28. doi: 10.1016/j.bbrep.2015.04.004. eCollection 2015 Jul. PMID: 29124142; PMCID: PMC5668623.
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Biochem Biophys Rep. 2015 Apr 25;2:23-28. doi: 10.1016/j.bbrep.2015.04.004. eCollection 2015 Jul.

High efficiency reduction capability for the formation of Fab׳ antibody fragments from F(ab).

Biochemistry and biophysics reports

Victor Crivianu-Gaita, Alexander Romaschin, Michael Thompson

Affiliations

  1. Chemistry Department, University of Toronto, 80 St. George Street, Toronto, ON, Canada M5S 3H6.
  2. Clinical Biochemistry, St. Michael?s Hospital, Toronto, ON, Canada M5B 1W8.

PMID: 29124142 PMCID: PMC5668623 DOI: 10.1016/j.bbrep.2015.04.004

Abstract

Antibodies have widespread applications in areas ranging from therapeutics to chromatography and protein microarrays. Certain applications require only the fragment antigen-binding (Fab) units of the protein. This study compares the cleavage efficacy of dithiothreitol (DTT), mercaptoethylamine (MEA), and dithiobutylamine (DTBA) - a relatively new reducing agent synthesized in 2012. Pseudo-first order kinetic analyses show DTBA to be ~213 times faster than DTT and ~71 times faster than MEA in the formation of Fab׳ antibody fragments from polyclonal rabbit antibodies. Monoclonal mouse antibodies were also used to show the feasibility of the reduction process on antibodies from a different species and with a different clonality. DTBA cleaved the monoclonal mouse F(ab)

Keywords: Antibody reduction; Dithiobutylamine; Dithiothreitol; Fab׳ fragments; Mercaptoethylamine

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