Pulsed ultrafiltration-mass spectrometry

Fig. 4.4 Scheme of pulsed ultrafiltration-mass spectrometry (PUF-MS) to screen chemical mixtures for compounds that bind to a macromolecular receptor. The ultrafiltration membrane traps a receptor in solution, but allows low molecular weight... 

Johnson BM, Nikolic D, van Breemen RB Applications of pulsed ultrafiltration-mass spectrometry. Mass Spectrom Rev 2002, 21, 76-86. 

Screening for inhibitors of dihydrofolate reductase using pulsed ultrafiltration mass spectrometry. Comb Chem High Throughput Screen 1998, 1, 47-55. 

Utility of pulsed ultrafiltration-mass spectrometry, where microsomal fractions are entrapped in stirred ultrafiltration chambers and the output of the chamber is introduced directly into an electrospray mass spectrometer shows particular promise [37]. 

Venton DL, Pulsed ultrafiltration mass spectrometry a new method for screening combinatorial libraries, Anal. Chem., 69 2159-2164, 1997. 

Pulsed Ultrafiltration-Mass Spectrometry. A versatile approach to screening solution phase combinatorial libraries and natural product extracts is pulsed ultrafiltration-mass spectrometry (61,62), which uses a standard LC-MS system with an ultrafiltration chamber substituted for the HPLC column. 

The second solution to increasing the throughput of pulsed ultrafiltration mass spectrometry has been to miniaturize the ultrafiltration chamber volume while maintaining the flow rate and chamber pressure. Because the ultrafiltration membrane cannot withstand high pressure without rupturing, the ultrafiltration process cannot be accelerated simply by increasing the flow rate through the chamber. The approach of Beverly et al. (72) was to fabricate a 35-/u,L ultra-... 

The advantages of pulsed ultrafiltration-mass spectrometry include the variety of different applications that may be carried out, the convenience of on-line screening, solution-phase screening, the ability to screen either combinatorial libraries or natural product extracts, the diversity of receptors that may be screened, and the freedom to use either volatile or non-volatile binding buffers. For metabolic and toxicity screening, flow injection analyses have the additional advantages that product feedback inhibition is prevented so that the metabolic profile more closely approximates the in vivo system ( 70). Finally, the... 

Van Breemen, R.B. Woodbury, C.P Venton, D.L. Screening Molecular Diversity Using Pulsed Ultrafiltration Mass Spectrometry, pp. 99-113 in Larsen, B.S. McEwen, C.N., eds.. Mass Spectrometry of Biological Materials, 2nd ed., Dekker, New York (1998). 

Van Breemen, R.B. Huang, C-R. Nikolic, D. Woodbury, C.P. Zhao, Y-Z. Venton, D.L. Pulsed Ultrafiltration Mass Spectrometry A New Method for Screening Combinatorial Libraries, Anal. Chem. 69, 2159-2164 (1997). 

Johnson, B.M. Nikolic, D. Van Breemen, R.B. Application of Pulsed Ultrafiltration-Mass Spectrometry, Mass Spectrom. Rev. 21, 76-86 (2002). 

Pulsed Ultrafiltration-Mass Spectrometry in this approach, the receptor was placed in an ultrafiltration membrane, and a pulse of library compounds was applied to this cell [104], The active components were retained in the ultrafiltration cell, and the unretained components and the binding buffer were washed away to the waste. The bound components were liberated and eluted from the membrane with a flow of methanol or an acidic solvent. ESI-MS or ESI-MS/MS provided the identity of the components liberated. 

Nikolic D, Fan PW, Bolton JL, van Breemen RB. Screening for xenobiotic electrophilic metabolites using pulsed ultrafiltration—mass spectrometry. Comb Chem High Throughput Screen 1999 2 165-175. 

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