Online coupling of immunoextraction, digestion, and microliquid chromatography-tandem mass spectrometry for the analysis of sarin and soman-butyrylcholinesterase adducts in human plasma

Bonichon, Maud; Valbi, Valentina; Combès, Audrey; Desoubries, Charlotte; Bossée, Anne; Pichon, Valérie

doi:10.1007/s00216-017-0640-z

Cited by 21 publications

(13 citation statements)

References 47 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…MS is increasingly important in identifying changes in the amino acid sequence of target proteins, resulting in many opportunities to discover useful biomarkers for prognosis, diagnosis and disease staging . The accurate analysis of intact molecular mass plays a crucial role in proteomic applications.…”

Section: Resultsmentioning

confidence: 99%

Identification of S419 on human serum albumin as a novel biomarker for sarin and cyclosarin exposure

et al. 2020

Rapid Comm Mass Spectrometry

View full text Add to dashboard Cite

Rationale:Organophosphorus nerve agents are highly toxic because they inhibit acetylcholinesterase activity, thereby causing a series of symptomatic poisoning.Upon entering the body, nerve agents bind active amino acid residues to form phosphonylated adducts. A potentially beneficial method for specific verification of exposure of nerve agents is based on albumin adducts, which have a half-life of 18 days. This appears to be more effective than the fluoride reactivation method, based on acetylcholinesterase.Methods: After the exposure of human serum albumin to nine nerve agents, human serum albumin was denatured, reduced, alkylated and digested with trypsin according to standard mass spectrometry-based proteomics procedures. The phosphonylated peptides of human serum albumin were identified using positive ion electrospray ionization with a quadrupole orbitrap mass spectrometer. Results:The peptide KVPQVSTPTLVESR showed a good mass spectrometric response to the nine nerve agents. The tendency of sarin and cyclosarin was to bind to S419 on the peptide, while the other nerve agents (tabun, soman and V-type nerve agents) were shown to bind more readily to K414 on the peptide. Conclusions:This research revealed a new site, S419, of the tryptic peptide KVPQVSTPTLVEVSR on human albumin to be a valuable biomarker for sarin/cyclosarin exposure, helping to further distinguish sarin and cyclosarin poisoning from that of other nerve agents and providing an important tool for the identification of sarin or cyclosarin in terrorist attacks.

show abstract

Section: Resultsmentioning

confidence: 99%

Identification of S419 on human serum albumin as a novel biomarker for sarin and cyclosarin exposure

et al. 2020

Rapid Comm Mass Spectrometry

View full text Add to dashboard Cite

show abstract

“…However, for some applications Hupresin outperforms procainamide-Sepharose. An example of the superior performance of Hupresin is the observation that HuBChE obtained by simply passing plasma over Hupresin was sufficiently purified to allow detection of the nerve agent modified active site peptide of HuBChE by mass spectrometry [41].…”

Section: Discussionmentioning

confidence: 99%

Purification of human butyrylcholinesterase from frozen Cohn fraction IV-4 by ion exchange and Hupresin affinity chromatography

et al. 2019

View full text Add to dashboard Cite

Human butyrylcholinesterase (HuBChE) is being developed as a therapeutic for protection from the toxicity of nerve agents. An enriched source of HuBChE is Cohn fraction IV-4 from pooled human plasma. For the past 40 years, purification of HuBChE has included affinity chromatography on procainamide-Sepharose. The present report supports a new affinity sorbent, Hupresin, for purification of HuBChE from Cohn fraction IV-4. Nine batches of 70–80 kg frozen Cohn fraction were extracted with water, filtered, and chromatographed on 30 L of Q-Ceramic ion exchange sorbent at pH 4.5. The 4% pure Q-eluent was pumped onto 4.2 L Hupresin, where contaminants were washed off with 0.3 M NaCl in 20 mM sodium phosphate pH 8.0, before 99% pure HuBChE was eluted with 0.1 M tetramethylammonium bromide. The average yield was 1.5 g of HuBChE from 80 kg Cohn paste. Recovery of HuBChE was reduced by 90% when the paste was stored at -20°C for 1 year, and reduced 100% when stored at 4°C for 24h. No reduction in HuBChE recovery occurred when paste was stored at -80°C for 3 months or 3 years. Hupresin and procainamide-Sepharose were equally effective at purifying HuBChE from Cohn fraction. HuBChE in Cohn fraction required 1000-fold purification to attain 99% purity, but 15,000-fold purification when the starting material was plasma. HuBChE (P06276) purified from Cohn fraction was a 340 kDa tetramer of 4 identical N-glycated subunits, stable for years in solution or as a lyophilized product.

show abstract

“…Research laboratories find Hupresin ® superior to the procainamide affinity gel for purifying BChE from human plasma and other sources [12, 15, 19]. Tables 2 and 3 list use of Hupresin ® and the procainamide affinity gel for purifying BChE from a variety of animal plasma, porcine milk and expression systems.…”

Section: Discussionmentioning

confidence: 99%

Purification of recombinant human butyrylcholinesterase on Hupresin®

Lockridge

David

Schopfer

et al. 2018

Journal of Chromatography B

View full text Add to dashboard Cite

Affinity chromatography on procainamide-Sepharose has been an important step in the purification of butyrylcholinesterase (BChE) and acetylcholinesterase (AChE) since its introduction in 1978. The procainamide affinity gel has limitations. In the present report a new affinity gel called Hupresin® was evaluated for its ability to purify truncated, recombinant human butyrylcholinesterase (rHuBChE) expressed in a stably transfected Chinese Hamster Ovary cell line. We present a detailed example of the purification of rHuBChE secreted into 3940 mL of serum-free culture medium. The starting material contained 13,163 units of BChE activity (20.9 mg). rHuBChE was purified to homogeneity in a single step by passage over 82 mL of Hupresin® eluted with 0.1 M tetramethylammonium bromide in 20 mM TrisCl pH 7.5. The fraction with the highest specific activity of 630 units/mg contained 11 mg of BChE. Hupresin® is superior to procainamide-Sepharose for purification of BChE, but is not suitable for purifying native AChE because Hupresin® binds AChE so tightly that AChE is not released with buffers, but is desorbed with denaturing solvents such as 50% acetonitrile or 1% trifluoroacetic acid. Procainamide-Sepharose will continue to be useful for purification of AChE.

show abstract

Online coupling of immunoextraction, digestion, and microliquid chromatography-tandem mass spectrometry for the analysis of sarin and soman-butyrylcholinesterase adducts in human plasma

Cited by 21 publications

References 47 publications

Identification of S419 on human serum albumin as a novel biomarker for sarin and cyclosarin exposure

Identification of S419 on human serum albumin as a novel biomarker for sarin and cyclosarin exposure

Purification of human butyrylcholinesterase from frozen Cohn fraction IV-4 by ion exchange and Hupresin affinity chromatography

Purification of recombinant human butyrylcholinesterase on Hupresin®

Contact Info

Product

Resources

About