Separation of enantiomers on a pepsin-bonded column

Chiral separation by chromatographic and electromigration techniques. A Review

“…Two cellobiohydrolases, CBH I and CBH II [222] and more recently CBH 58 [223], found application as chiral selectors for the preparation of silica-based CSPs which showed remarkable enantioselectivity for a broad spectrum of drugs, among them b-blockers [224]. Lysozyme [225]-and pepsin [226]-based CSPs showed enantioselectivity for basic and neutral enantiomers but not for acidic enantiomers. Recently, the preparation of amyloglucosidasebased CSPs and its application to aminoalcohols was reported [227].…”

Chiral Recognition Mechanism: Practical Considerations for Pharmaceutical Analysis of Chiral Compounds

“…A few classes of proteins including albumins, glycoproteins, and enzymes have been chemically bonded to silica gel and evaluated as HPLC CSPs [17]. Among them, CSPs based on bovine serum albumin (BSA) [86], human serum albumin (HSA) [87], α-acid glycoprotein (AGP) [88], ovomucoid (OVM) from chicken egg whites [89], avidin (AVI) [90], cellobiohydrolase I (CBH I) [91], and pepsin [92] are commercially available. Some important physical properties and applicability of these commercialized CSPs are given in Table 1.…”

HPLC Chiral Stationary Phases Produced with Isolated Human Serum Albumin Fragments

“…This property has led to the development of a number of HPLC chiral stationary phases based upon albumins such as bovine serum albumin (BSA) 1 and human serum albumin (HSA), 2 glycoproteins such as α1-acid glycoprotein, 3,4 ovomucoid, 5 ovoglycoprotein, 6 avidin 7 and cellulase 8 and enzymes such as trypsin, 9 α-chymotrypsin, 10 lysozyme 11 and pepsin. 12 Disadvantages of protein-based HPLC columns have included low capacity, lack of column ruggedness in some instances and a limited understanding of the chiral recognition mechanisms. If a chiral binding site(s) exists on a domain or fragment and if it acts independently of others, chiral columns with the desired domain or fragment should be able to be made; these could be of higher capacity and more stable.…”