Papain digestion of different mouse IgG subclasses as studied by electrospray mass spectrometry

被引:43
作者
Adamczyk, M [1 ]
Gebler, JC [1 ]
Wu, J [1 ]
机构
[1] Abbott Labs, Abbott Diagnost Div, Dept Chem 9NM, Abbott Pk, IL 60064 USA
关键词
IgG; papain digestion; subclass; electrospray mass spectrometry;
D O I
10.1016/S0022-1759(00)00135-6
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
On-line liquid chromatography-electrospray ionization mass spectrometry (LC/ESI-MS) has been utilized to monitor the papain digestion of mouse monoclonal antibodies (mAb) of different subclasses (IgG1, IgG2a, IgG2b, and IgG3). The method is simple, rapid, sensitive, and allows for simultaneous determination of digestion products and the identification of microheterogenous fragments. The results provide important insight into the mechanism of papain digestion of mouse IgGs of different subclasses. (C) 2000 Elsevier Science B.V. All rights reserved.
引用
收藏
页码:95 / 104
页数:10
相关论文
共 33 条
[21]   MONOCLONAL-ANTIBODIES - PURIFICATION, FRAGMENTATION AND APPLICATION TO STRUCTURAL AND FUNCTIONAL-STUDIES OF CLASS-I MHC ANTIGENS [J].
PARHAM, P ;
ANDROLEWICZ, MJ ;
BRODSKY, FM ;
HOLMES, NJ ;
WAYS, JP .
JOURNAL OF IMMUNOLOGICAL METHODS, 1982, 53 (02) :133-173
[22]  
PARHAM P, 1986, HDB EXPT IMMUNOLOGY, V1
[23]  
PRASAD L, 1988, J BIOL CHEM, V263, P2571
[24]   A NOVEL METHOD FOR CONTROLLING THE PEPSIN DIGESTION OF ANTIBODIES [J].
REA, DW ;
ULTEE, ME .
JOURNAL OF IMMUNOLOGICAL METHODS, 1993, 157 (1-2) :165-173
[25]   AN INTEGRATED STRATEGY FOR STRUCTURAL CHARACTERIZATION OF THE PROTEIN AND CARBOHYDRATE COMPONENTS OF MONOCLONAL-ANTIBODIES - APPLICATION TO ANTI-RESPIRATORY SYNCYTIAL VIRUS MAB [J].
ROBERTS, GD ;
JOHNSON, WP ;
BURMAN, S ;
ANUMULA, KR ;
CARR, SA .
ANALYTICAL CHEMISTRY, 1995, 67 (20) :3613-3625
[26]   THE DIFFERENTIAL ENZYME SENSITIVITY OF RAT IMMUNOGLOBULIN-G SUBCLASSES TO PAPAIN AND PEPSIN [J].
ROUSSEAUX, J ;
BISERTE, G ;
BAZIN, H .
MOLECULAR IMMUNOLOGY, 1980, 17 (04) :469-482
[27]  
ROUSSEAUX J, 1986, METHOD ENZYMOL, V121, P663
[28]   OPTIMAL CONDITIONS FOR THE PREPARATION OF FAB AND F(AB')2 FRAGMENTS FROM MONOCLONAL IGG OF DIFFERENT RAT IGG SUBCLASSES [J].
ROUSSEAUX, J ;
ROUSSEAUXPREVOST, R ;
BAZIN, H .
JOURNAL OF IMMUNOLOGICAL METHODS, 1983, 64 (1-2) :141-146
[29]  
ROUSSEAUX J, 1990, RAT HYBRIDOMAS RAT M, P210
[30]  
Siegel M M, 1996, Methods Mol Biol, V61, P211