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Spurious capillary zone electrophoresis pattern in hypercholesterolemic dogs.

ABSTRACT: Capillary zone electrophoresis (CZE) is a relatively new serum protein electrophoresis method with higher resolution than other electrophoretic techniques. Hypercholesterolemic dogs exhibit a peculiar CZE pattern. Specifically, they have a shoulder or peak immediately next to the albumin peak. We investigated the prevalence of this spurious peak in hypercholesterolemic dogs and its correlation with the serum cholesterol concentration. Moreover, possible discrepancies between the CZE and spectrophotometric (bromocresol green [BCG] method) albumin concentrations in those animals were evaluated, as well as the accuracy in measuring albumin by a different CZE fractionation system. We retrospectively enrolled 500 hypercholesterolemic and normotriglyceridemic dogs. Each electrophoretic curve was inspected visually to identify a spurious peak (prevalence of 68.8%). We chose 120 dogs to further investigate the albumin concentration; CZE albumin was significantly higher than measured using the BCG method. A weak but significant correlation (r = 0.412; p <0.0001) was observed between the magnitude of the spurious peak and the serum cholesterol concentration. Finally, the significant difference between CZE and BCG albumin measurement disappeared (p = 0.92) when the spurious peak was considered as α1-globulins instead of albumin.

SUBMITTER: Mangiagalli G

PROVIDER: S-EPMC9999394 | biostudies-literature | 2023 Mar

REPOSITORIES: biostudies-literature

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Spurious capillary zone electrophoresis pattern in hypercholesterolemic dogs.

Mangiagalli Giulia G Meazzi Sara S Giordano Alessia A Rossi Silvia S

Journal of veterinary diagnostic investigation : official publication of the American Association of Veterinary Laboratory Diagnosticians, Inc 20221207 2

Capillary zone electrophoresis (CZE) is a relatively new serum protein electrophoresis method with higher resolution than other electrophoretic techniques. Hypercholesterolemic dogs exhibit a peculiar CZE pattern. Specifically, they have a shoulder or peak immediately next to the albumin peak. We investigated the prevalence of this spurious peak in hypercholesterolemic dogs and its correlation with the serum cholesterol concentration. Moreover, possible discrepancies between the CZE and spectrop ...[more]

PMID: 36476172

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Project description:Out of all categories, monoclonal antibody (mAb) therapeutics attract the most interest due to their strong therapeutic potency and specificity. Six of the 10 top-selling drugs are antibody-based therapeutics that will lose patent protection soon. The European Medicines Agency has pioneered the regulatory framework for approval of biosimilar products and approved the first biosimilar antibodies by the end of 2013. As highly complex glycoproteins with a wide range of micro-variants, mAbs require extensive characterization through multiple analytical methods for structure assessment rendering manufacturing control and biosimilarity studies particularly product and time-consuming. Here, capillary zone electrophoresis coupled to mass spectrometry by a sheathless interface (CESI-MS) was used to characterize marketed reference mAbs and their respective biosimilar candidate simultaneously over different facets of their primary structure. CESI-MS/MS data were compared between approved mAbs and their biosimilar candidates to prove/disconfirm biosimilarity regarding recent regulation directives. Using only a single sample injection of 200 fmol, CESI-MS/MS data enabled 100% amino acids (AA) sequence characterization, which allows a difference of even one AA between 2 samples to be distinguished precisely. Simultaneously glycoforms were characterized regarding their structures and position through fragmentation spectra and glycoforms semiquantitative analysis was established, showing the capacity of the developed methodology to detect up to 16 different glycans. Other posttranslational modifications hotspots were characterized while their relative occurrence levels were estimated and compared to biosimilars. These results proved the value of using CESI-MS because the separation selectivity and ionization efficiency provided by the system allowed substantial improvement in the characterization workflow robustness and accuracy. Biosimilarity assessment could be performed routinely with a single injection of each candidate enabling improvements in the biosimilar development pipeline.

Dataset Information

Spurious capillary zone electrophoresis pattern in hypercholesterolemic dogs.

Publications

Spurious capillary zone electrophoresis pattern in hypercholesterolemic dogs.

Similar Datasets

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