Region-Selective Labeling of Antibodies as Determined by Electrospray Ionization-Mass Spectrometry (ESI-MS)
2000; American Chemical Society; Volume: 11; Issue: 4 Linguagem: Inglês
10.1021/bc990181y
ISSN1520-4812
AutoresMaciej Adamczyk, John C. Gebler, Kevin Shreder, Jiang Wu,
Tópico(s)Microfluidic and Capillary Electrophoresis Applications
ResumoThe electrospray ionization-mass spectrometry (ESI-MS) analysis of three sets of monoclonal antibody-acridinium-9-carboxamide conjugates is described. The conjugates (nine total) were enzymatically digested using papain and the resulting fragments [Fc heavy chain, Fab, or F(ab')(2)] were analyzed using liquid chromatography/ESI-MS. The average number of labels per fragment were calculated using Sigma nx%, where n is the number of acridinium molecules covalently bound to the fragment and x% is the percent relative area of the corresponding peaks in the mass spectrum. When these values were normalized against the molecular weight of their respective region, antibody-dependent labeling patterns were observed. For antibodies T (anti-L-T(4)) and F (anti-FITC), there was a preference for conjugation of the Fab region over the Fc region. For antibody B (anti-biotin), the trend was reversed.
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