Aminopeptidase

Manufactured by Merck Group

Sourced in Germany

Aminopeptidase is a type of hydrolytic enzyme that cleaves amino acids from the N-terminal end of polypeptide chains. It plays a key role in the breakdown and metabolism of proteins. Aminopeptidase is used in various laboratory applications for the analysis and characterization of protein samples.

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2 protocols using aminopeptidase

Quantification of Maillard Reaction Products

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Acetonitrile (LC-MS grade) and methanol (HPLC grade) were obtained from VWR Prolabo (Darmstadt, Germany). Freshly double distilled water (Bi 18E double distillation system, QCS, Maintal, Germany) was used for solvents production for LC-MS analysis. Nonafluoropentanoic acid (NFPA) was from Sigma-Aldrich (Steinheim, Germany). Reference material for calibration of MRPs was synthesized in our laboratory as described before: N-ε-fructosyllysine [30 (link)], pyrraline [31 (link),32 (link)], CML [17 (link)], CEL [17 (link)], and MG-H1 [17 (link)]. Stable isotope labelled internal standards for HPLC-MS/MS analysis were synthesized similarly but using [¹³C₆,¹⁵N₂]lysine ([¹³C₆,¹⁵N₂]Pyr) and [¹³C₆]arginine ([¹³C₆]MG-H1) instead of the unlabeled compounds. The synthesis of [¹³C₃]CEL was described recently [29 (link)]. [²H₂]CML was purchased from PolyPeptide (Strasbourg, France), [¹³C₆,¹⁵N₂]lysine from Campro (Berlin, Germany), and [¹³C₆]arginine from Eurisotop (Saarbrücken, Germany). Enzymatic hydrolysis was performed by using pronase E (Merck, Darmstadt, Germany) and pepsin, aminopeptidase and prolidase (all from Sigma-Aldrich, Steinheim, Germany).

Manig F., Hellwig M., Pietz F, & Henle T. (2022). Studies about the Dietary Impact on “Free” Glycation Compounds in Human Saliva. Foods, 11(14), 2112.

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Recombinant GRFT Protein Preparation

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Recombinant GRFT drug substance was supplied by Kentucky Bioprocessing LLC (Owensboro, KY). The protein was supplied in a solution of PBS (12 mg/mL for 2-year stability study; 10 mg/mL for all other assessments). PBS 10× molecular biology grade (pH 7.4) was purchased from Mediatech, Inc. (Manassa, VA). Acetonitrile (ACN), trifluoroacetic acid (TFA), sodium acetate, formic acid, phosphate acid, potassium phosphate monobasic, potassium phosphate dibasic, Tris, hydrochloric acid (HCl), and sodium hydroxide (NaOH) were obtained from Fisher Scientific (Pittsburgh, PA). Hydrogen peroxide (H₂O₂, 30%), disodium ethylenediaminetetraacetic acid (EDTA), d-α tocopheryl polyethylene glycol 1000 succinate (vitamin E TPGS), ascorbic acid, methionine, glutathione, cysteine, aminopeptidase, lysozyme, and proteinase K were obtained from Sigma (St. Louis, MO). A MilliQ (Millipore; Milford, MA) water filtration system operating at 18.2 MΩ cm was used for water.

Kramzer L.F., Hamorsky K.T., Graebing P.W., Wang L., Fuqua J.L., Matoba N., Lasnik A.B., Moncla B.J., Zhang J., Palmer K.E, & Rohan L.C. (2021). Preformulation Characterization of Griffithsin, a Biopharmaceutical Candidate for HIV Prevention. AAPS PharmSciTech, 22(3), 83.

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