Purification, characterization, and N-glycosylation of recombinant butyrylcholinesterase from transgenic rice cell suspension cultures

Jasmine M. Corbin, Muchena J. Kailemia, C. Linn Cadieux, Salem Alkanaimsh, Kalimuthu Karuppanan, Raymond L. Rodriguez, Carlito B Lebrilla, Douglas M. Cerasoli, Karen A. McDonald, Somen Nandi

Research output: Contribution to journalArticle

4 Citations (Scopus)

Abstract

Recombinant butyrylcholinesterase produced in a metabolically regulated transgenic rice cell culture (rrBChE) was purified to produce a highly pure (95%), active form of enzyme. The developed downstream process uses common manufacturing friendly operations including tangential flow filtration, anion-exchange chromatography, and affinity chromatography to obtain a process recovery of 42% active rrBChE. The purified rrBChE was then characterized to confirm its comparability to the native human form of the molecule (hBChE). The recombinant and native enzyme demonstrated comparable enzymatic behavior and had an identical amino acid sequence. However, rrBChE differs in that it contains plant-type complex N-glycans, including an α-1,3 linked core fucose, and a β-1,2 xylose, and lacking a terminal sialic acid. Despite this difference, rrBChE is demonstrated to be an effective stoichiometric bioscavenger for five different organophosphorous nerve agents in vitro. Together, the efficient downstream processing scheme and functionality of rrBChE confirm its promise as a cost-effective alternative to hBChE for prophylactic and therapeutic use.

Original languageEnglish (US)
Pages (from-to)1301-1310
Number of pages10
JournalBiotechnology and Bioengineering
Volume115
Issue number5
DOIs
StatePublished - May 1 2018

Fingerprint

Glycosylation
Butyrylcholinesterase
Purification
Suspensions
Enzymes
Cell Culture Techniques
Affinity chromatography
Xylose
Fucose
N-Acetylneuraminic Acid
Therapeutic Uses
Chromatography
Affinity Chromatography
Population Groups
Cell culture
Anions
Polysaccharides
Amino acids
Amino Acid Sequence
Ion exchange

Keywords

  • butyrylcholinesterase
  • downstream processing
  • Hupresin
  • N-glycosylation

ASJC Scopus subject areas

  • Biotechnology
  • Bioengineering
  • Applied Microbiology and Biotechnology

Cite this

Corbin, J. M., Kailemia, M. J., Cadieux, C. L., Alkanaimsh, S., Karuppanan, K., Rodriguez, R. L., ... Nandi, S. (2018). Purification, characterization, and N-glycosylation of recombinant butyrylcholinesterase from transgenic rice cell suspension cultures. Biotechnology and Bioengineering, 115(5), 1301-1310. https://doi.org/10.1002/bit.26557

Purification, characterization, and N-glycosylation of recombinant butyrylcholinesterase from transgenic rice cell suspension cultures. / Corbin, Jasmine M.; Kailemia, Muchena J.; Cadieux, C. Linn; Alkanaimsh, Salem; Karuppanan, Kalimuthu; Rodriguez, Raymond L.; Lebrilla, Carlito B; Cerasoli, Douglas M.; McDonald, Karen A.; Nandi, Somen.

In: Biotechnology and Bioengineering, Vol. 115, No. 5, 01.05.2018, p. 1301-1310.

Research output: Contribution to journalArticle

Corbin, JM, Kailemia, MJ, Cadieux, CL, Alkanaimsh, S, Karuppanan, K, Rodriguez, RL, Lebrilla, CB, Cerasoli, DM, McDonald, KA & Nandi, S 2018, 'Purification, characterization, and N-glycosylation of recombinant butyrylcholinesterase from transgenic rice cell suspension cultures', Biotechnology and Bioengineering, vol. 115, no. 5, pp. 1301-1310. https://doi.org/10.1002/bit.26557
Corbin, Jasmine M. ; Kailemia, Muchena J. ; Cadieux, C. Linn ; Alkanaimsh, Salem ; Karuppanan, Kalimuthu ; Rodriguez, Raymond L. ; Lebrilla, Carlito B ; Cerasoli, Douglas M. ; McDonald, Karen A. ; Nandi, Somen. / Purification, characterization, and N-glycosylation of recombinant butyrylcholinesterase from transgenic rice cell suspension cultures. In: Biotechnology and Bioengineering. 2018 ; Vol. 115, No. 5. pp. 1301-1310.
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