Potency/Quantity

Protein Concentration by Bradford, BCA, or Lowry

Protein detection and quantification using Bradford, BCA or Lowry Protein quantification Protease Assay Kit is employed. These cuvette or microwell based assays are tested in conjunction with the client’s sample to detect and quantify all levels and forms of protein. Concentration is reported in mass per volume sample. Use of these methods for specific applications will be evaluated for repeatability, linearity, LOD/LOQ and accuracy.

• Sample required: microgram to milligram amounts of protein

Amino Acid Analysis

Amino Acid Analysis is performed according to Analysis #10030. The assay is carried out in two steps: 1) acid hydrolysis (usually hydrochloric acid) of protein into component amino acids using the validated Eldex Hydrolysis/Derivatization Workstation; 2) Derivatized the amino acids using the Waters AccQTag Chemistry. Eldex Hydrolysis/Derivatization Workstation Overview: This system is designed to provide convenient hydrolysis and pre-column derivatization as it applies to protein hydrolysis and amino acid modifications performed in the Biopharmaceutical Services area. Typically 6N hydrochloric acid at 110 deg C for 24 h is used to hydrolyze the protein and then it is removed under reduced pressure prior to derivatization. A Waters UPLC is used to separate the derivatized amino acids and from standard plots, the unknown amino acid concentration is quantified. We will evaluate the examine linearity, accuracy, and the matrix for interferences. We will report the amino acid composition in terms of number of residues, mol% or concentration of protein. One week of development will allow for two full runs of the analysis.

• Sample required: microgram to milligram amounts of protein