Combining Avidin−Biotin Chemistry with Matrix-Assisted Laser Desorption/Ionization Mass Spectrometry

• Published in: Analytical chemistry (Washington) Vol. 68; no. 19; pp. 3382 - 3387
• Main Authors: Schriemer, David C, Li, Liang
• Format: Journal Article
• Language: English
• Published: Washington, DC American Chemical Society 01.10.1996
• Subjects: Analytical biochemistry: general aspects, technics, instrumentation; Analytical, structural and metabolic biochemistry; Avidin - chemistry; Biological and medical sciences; Biotin - chemistry; Chemistry; Fundamental and applied biological sciences. Psychology; Molecular Structure; Scientific imaging; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization - methods; Biochemical analysis; Vitamin; Glycoproteins; Biotin; Mass spectrometry; Avidin; Laser desorption; Chemical ionization
• ISSN: 0003-2700; 1520-6882
• DOI: 10.1021/ac960219p

Molecular recognition based on the high affinity of avidin for biotin has been combined with matrix-assisted laser desorption/ionization (MALDI) mass spectrometry. The rich chemistry of biotinylation reagents underlies the development of a mass tracer method for the rapid and sensitive analysis of biotinylated analyte in complex mixtures. In this method, the biotinylated analyte is captured with immobilized avidin agarose beads. The beads with the bound complex are deposited on the MALDI sample probe, followed by elution of the retained compounds and removal of the beads. The use of MALDI matrix solutions in conjunction with drying of the beads is shown to be sufficient to overcome the avidin−biotin interaction. The studies of several different avidin−biotin interaction schemes tailored to the MALDI analysis are presented. It is demonstrated that, with an optimized scheme, the extremely high selectivity of the avidin−biotin interaction is preserved, generating species represented in the MALDI spectra that arise only from the avidin−biotin interaction. Biotinylated species in the nanomolar range can be isolated and analyzed with this technique.