DIRECT ANALYSIS OF AFFINITY-BOUND ANALYTES BY MALDI/TOF MS

被引：74

作者：

PAPAC, DI

HOYES, J

TOMER, KB

机构：

[1] NIEHS,MOLEC BIOPHYS LAB,RES TRIANGLE PK,NC 27709

[2] VG ANALYT,MANCHESTER M23 9LE,LANCS,ENGLAND

来源：

ANALYTICAL CHEMISTRY | 1994年 / 66卷 / 17期

关键词：

D O I：

10.1021/ac00089a004

中图分类号：

O65 [分析化学];

学科分类号：

070302 ; 081704 ;

摘要：

Identification of ligands separated with affinity chromatography has been facilitated by direct analysis of the bound ligand using matrix-assisted laser desorption time-of-flight mass spectrometry. Tbe mass spectral detection of analytes separated by immunoaffinity chromatography and immobilized metal ion affinity chromatography is shown. For example, cytochrome c is used as an affinity support to purify the anti-cytochrome c monoclonal antibody from ascites, and the mass spectrum of the anti-cytochrome c monoclonal antibody was obtained by direct analysis of an aliquot of the column bed. Direct analyses of metal binding proteins and phosphopeptides bound to immobilized metal ion affinity columns are also demonstrated. The method is characterized by minimal sample manipulation and high sensitivity with low picomoles and high femtomoles of analytes being readily observed.

引用

收藏

页码：2609 / 2613

页数：5

相关论文

共 12 条

[1] ISOLATION OF PHOSPHOPROTEINS BY IMMOBILIZED METAL (FE-3+) AFFINITY-CHROMATOGRAPHY [J].

ANDERSSON, L ;

PORATH, J .

ANALYTICAL BIOCHEMISTRY, 1986, 154 (01) :250-254

[2] PURIFICATION OF TRANSFERRIN AND ALBUMIN FROM MOUSE ASCITES-FLUID [J].

FELDHOFF, RC ;

STEFFEN, MC ;

GEOGHEGAN, TE ;

LEDFORD, BE .

PREPARATIVE BIOCHEMISTRY, 1985, 15 (04) :221-236

[3] NEW DESORPTION STRATEGIES FOR THE MASS-SPECTROMETRIC ANALYSIS OF MACROMOLECULES [J].

HUTCHENS, TW ;

YIP, TT .

RAPID COMMUNICATIONS IN MASS SPECTROMETRY, 1993, 7 (07) :576-580

[4] MATRIX-ASSISTED LASER DESORPTION IONIZATION MASS-SPECTROMETRY [J].

KARAS, M ;

BAHR, U ;

GIESSMANN, U .

MASS SPECTROMETRY REVIEWS, 1991, 10 (05) :335-357

[5]

KLINE T, 1983, HDB AFFINITY CHROMAT, V63

[6] EXPRESSION OF GLYCOSYLATED AND NONGLYCOSYLATED HUMAN TRANSFERRIN IN MAMMALIAN-CELLS - CHARACTERIZATION OF THE RECOMBINANT PROTEINS WITH COMPARISON TO 3 COMMERCIALLY AVAILABLE TRANSFERRINS [J].

MASON, AB ;

MILLER, MK ;

FUNK, WD ;

BANFIELD, DK ;

SAVAGE, KJ ;

OLIVER, RWA ;

GREEN, BN ;

MACGILLIVRAY, RTA ;

WOODWORTH, RC .

BIOCHEMISTRY, 1993, 32 (20) :5472-5479

[7] STRUCTURAL STUDY OF THE SUGAR CHAINS OF HUMAN LACTOFERRIN - FINDING OF 4 NOVEL COMPLEX-TYPE ASPARAGINE-LINKED SUGAR CHAINS [J].

MATSUMOTO, A ;

YOSHIMA, H ;

TAKASAKI, S ;

KOBATA, A .

JOURNAL OF BIOCHEMISTRY, 1982, 91 (01) :143-155

[8]

MICHEL H, 1988, J BIOL CHEM, V263, P1123

[9] LASER-DESORPTION TIME-OF-FLIGHT MASS-SPECTROMETRIC ANALYSIS OF TRANSFERRIN PRECIPITATED WITH ANTISERUM - A UNIQUE SIMPLE METHOD TO IDENTIFY MOLECULAR-WEIGHT VARIANTS [J].

NAKANISHI, T ;

OKAMOTO, N ;

TANAKA, K ;

SHIMIZU, A .

BIOLOGICAL MASS SPECTROMETRY, 1994, 23 (04) :230-233

[10] ELECTROSPRAY-IONIZATION MASS-SPECTROMETRY OF PHOSPHOPEPTIDES ISOLATED BY ONLINE IMMOBILIZED METAL-ION AFFINITY-CHROMATOGRAPHY [J].

NUWAYSIR, LM ;

STULTS, JT .

JOURNAL OF THE AMERICAN SOCIETY FOR MASS SPECTROMETRY, 1993, 4 (08) :662-669