Rapid LC-TOFMS method for identification of binding sites of covalent acylglucuronide–albumin complexes

Characterization of the binding epitope of ciprofloxacin bound to human serum albumin

“…As a major factor the reversible noncovalent binding to human serum albumin (HSA) has to be tested. The protein binding has been studied by a number of techniques such as electrophoresis [1], chromatography [2], crystallography [3], mass spectrometry [4], ultrafiltration [5] and NMR [6]. HSA exhibits six binding sites three of which are high-affinity sites [7][8][9].…”

Electrospray ionization mass spectrometry as a tool for determination of drug binding sites to human serum albumin by noncovalent interaction

“…Site I binds compounds like warfarin and azapropazone, site II has a high affinity for indole and benzodiazepine‐like compounds, site III binds digitoxin, biliary acids and aspirin, site IV binds billirubin, site V binds fatty acids, and finally site VI binds metal ions 4. Ligand binding to HSA has been studied by numerous techniques such as affinity capillary electrophoresis (ACE),5 ultracentrifugation,6 ultrafiltration,7 gel chromatography,8 gel filtration,9 high‐performance liquid chromatography (HPLC),10 crystallography,11 fluorescence spectroscopy,12 nuclear magnetic resonance (NMR),13 surface plasmon resonance,14 and combinations of these techniques 15…”

Development of an automated on-line pepsin digestion–liquid chromatography–tandem mass spectrometry configuration for the rapid analysis of protein adducts of chemical warfare agents

“…For example, a nerve agent exposure will result in the formation of adducts to human butyryl cholinesterase (HuBuChE) [5], whereas the vesicant sulfur mustard will give rise to rather random adducts, e.g., to human serum albumin (HSA) [6][7][8]. Similar covalent protein adducts are often observed in drug toxicology (see, e.g., [9][10][11]), in metabolism (e.g., [12][13][14][15][16]) and from post-translational modification (e.g., [17,18]). The analysis of covalent protein adducts is usually performed by enzymatic digestion of the modified protein and subsequent mass spectrometric analysis of the resultant peptide or amino acid adducts.…”