On-line continuous-flow, multi-protein biochemical assays for the characterization of bioaffinity compounds using electrospray quadrupole time-of-flight mass spectrometry

R.J.E. Derks, A.C. Hogenboom, G. van der Zwan, H. Irth

Research output: Contribution to JournalArticleAcademicpeer-review

Abstract

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 fluoresceinbiotin/streptavidin and digoxin/anti-digoxigenin.
Original languageEnglish
Pages (from-to)3376-84
JournalAnalytical Chemistry
Volume75
Issue number14
DOIs
Publication statusPublished - 2003

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