On-Line Continuous-Flow, Multi-Protein Biochemical Assays for the Characterization of Bioaffinity Compounds Using Electrospray Quadrupole Time-of-Flight Mass Spectrometry

The applicability of a homogeneous on-line continuous-flow, multi-protein biochemical assay was demonstrated for the interaction between fluorescein−biotin and streptavidin and for digoxin and anti-digoxigenin using electrospray quadrupole time-of-flight mass spectrometry (Q-TOF MS). In the on-line...

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Bibliographic Details
Published in:Analytical chemistry (Washington) 2003-07, Vol.75 (14), p.3376-3384
Main Authors: Derks, R. J. E, Hogenboom, A. C, van der Zwan, G, Irth, H
Format: Article
Language:English
Subjects:
Algorithms
Analytical biochemistry: general aspects, technics, instrumentation
Analytical chemistry
Analytical, structural and metabolic biochemistry
Biological and medical sciences
Buffers
Chemistry
Combinatorial Chemistry Techniques
Digoxigenin - analysis
Digoxin - analysis
Exact sciences and technology
Fundamental and applied biological sciences. Psychology
Ligands
Online Systems
Proteins - chemistry
Spectrometric and optical methods
Spectrometry, Mass, Electrospray Ionization
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Summary:The applicability of a homogeneous on-line continuous-flow, multi-protein biochemical assay was demonstrated for the interaction between fluorescein−biotin and streptavidin and for digoxin and anti-digoxigenin using electrospray quadrupole time-of-flight mass spectrometry (Q-TOF MS). In the on-line continuous-flow biochemical MS-based system several receptors (e.g., streptavidin and anti-digoxigenin, respectively) were allowed to react with corresponding reporter ligands (e.g.,fluorescein−biotin and digoxin, respectively). The methodology presented allows the simultaneous measurement of affinity and molecular mass of an active compound. By using automated MS and MS−MS switching functions of the Q-TOF, structure information is obtained allowing the characterization of bioactive compounds. No cross-reactivities were observed between the two model systems fluorescein−biotin/streptavidin and digoxin/anti-digoxigenin.
ISSN:0003-2700
1520-6882
DOI:10.1021/ac026423w