Characterisation of TNF-α-related peptides by high-performance liquid chromatography-mass spectrometry and high-performance liquid chromatography-tandem mass spectrometry

Alewood P.F., Bailey A.J., Brinkworth R.I., Fairlie D. and Jones A. (1993) Characterisation of TNF-α-related peptides by high-performance liquid chromatography-mass spectrometry and high-performance liquid chromatography-tandem mass spectrometry. Journal of Chromatography A, 646 1: 185-191. doi:10.1016/S0021-9673(99)87020-4


Author Alewood P.F.
Bailey A.J.
Brinkworth R.I.
Fairlie D.
Jones A.
Title Characterisation of TNF-α-related peptides by high-performance liquid chromatography-mass spectrometry and high-performance liquid chromatography-tandem mass spectrometry
Journal name Journal of Chromatography A   Check publisher's open access policy
ISSN 0021-9673
Publication date 1993-08-27
Sub-type Article (original research)
DOI 10.1016/S0021-9673(99)87020-4
Volume 646
Issue 1
Start page 185
End page 191
Total pages 7
Subject 1602 Criminology
1308 Clinical Biochemistry
1313 Molecular Medicine
Abstract Ionspray triple quadrupole mass spectrometry and high-performance liquid chromatography were used to investigate the products from the solid phase synthesis of (H)-Leu-Thr-Glu-Asn-(OH), a TNF-α active-site probe. The target sequence was assembled using tert.-butoxycarbonyl (Boc) chemistry in stepwise fashion from the C-terminal on an Boc-Asn-OCH2-Pamcopoly) (styrene-divinylbenzene) resin [Pam = 4-(carboxamidomethyl)benzyl ester]. The crude product was deprotected and cleaved from the resin by HF-p-cresol treatment for 1 h at 0°C. HPLC analysis at 214 nm indicated two late-eluting major products and an early-eluting product. Preparative HPLC demonstrated that the early-eluting product contained ca. 80% of the expected recovered sample mass. Each component was then directly analysed spectrometry and tandem mass spectrometry. The early eluting peak was confirmed as the LTEN sequence. Synthesis of the same sequence using 9-fluorenyl methoxycarbonly (Fmoc) chemistry gave an identical product and comfirmed the above analysis. The most significant by-product was derived from arylation of the glutamyl group by the quencher p-cresol. The likely origins of the by-products are discussed.
Q-Index Code C1
Institutional Status Unknown

Document type: Journal Article
Sub-type: Article (original research)
Collection: Scopus Import - Archived
 
Versions
Version Filter Type
Citation counts: Scopus Citation Count Cited 2 times in Scopus Article | Citations
Google Scholar Search Google Scholar
Created: Tue, 13 Sep 2016, 10:34:12 EST by System User