COMPARISON OF AGAROSE GEL AND CAPILLARY ZONE ELECTROPHORESIS METHODS USING PLASMA FROM GREEN TURTLES (CHELONIA MYDAS)

Abstract: Agarose gel electrophoresis (AGE) has been widely implemented throughout veterinary medicine and for analysis of plasma proteins of avian and reptile species. Capillary zone electrophoresis (CZE) is becoming a standard method in human clinical pathology laboratories but has not widely been used for the analysis of animal samples. The objective of the present study was to compare protein fractions derived from AGE and CZE methods using plasma from the green turtle (Chelonia mydas). Plasma samples were analyzed by AGE and CZE per manufacturer guidelines. The methods were assessed by CV analysis, Spearman's correlation, Passing-Bablok regression, and Bland Altman plots. CZE consistently resolved more fractions than AGE with three fractions observed in the prealbumin migrating region versus one for AGE and two fractions in the γ globulin region versus one for AGE. Compared with AGE, CZE showed a lower CV in intra-assay tests (1.0–4.9% vs 2.0–28.3%) and a lower or overlapping CV in interassay tests (1.0–10.6 vs 2.3–22.0). The prealbumin, α2 globulin, and β globulin fractions correlated the least between the methods (for all three fractions: rs ≤ 0.28, P > 0.21). Moderate, significant correlations between AGE and CZE methods were observed for albumin (rs = 0.78, P < 0.0001) and γ globulins (rs = 0.78, P < 0.0001). CZE has a higher precision and ease of use over AGE and offers the opportunity to resolve additional protein fractions. This will necessitate the development of new conventions in placement of fraction delimits, definition of species-specific reference intervals, and evaluation of clinical utility in abnormal turtles.

[1]  R. Marschang,et al.  REFERENCE INTERVALS FOR PLASMA CAPILLARY ZONE ELECTROPHORESIS IN HERMANN'S TORTOISES (TESTUDO HERMANNI) DEPENDING ON SEASON AND SEX , 2019, Journal of Zoo and Wildlife Medicine.

[2]  A. Regeniter,et al.  Peaks and tails: Evaluation of irregularities in capillary serum protein electrophoresis. , 2018, Clinical biochemistry.

[3]  G. Piccione,et al.  Influence of short-term storage conditions on the stability of total protein concentrations and electrophoretic fractions in plasma samples from loggerhead sea turtles, Caretta caretta , 2015, Comparative Clinical Pathology.

[4]  K. Arheart,et al.  ASSESSMENT OF HEMOGLOBIN BINDING PROTEIN IN LOGGERHEAD SEA TURTLES (CARETTA CARETTA) UNDERGOING REHABILITATION , 2014, Journal of zoo and wildlife medicine : official publication of the American Association of Zoo Veterinarians.

[5]  M. S. Jalme,et al.  Plasma Protein Electrophoresis in Birds: Comparison of a Semiautomated Agarose Gel System With an Automated Capillary System , 2013, Journal of avian medicine and surgery.

[6]  S. Paltrinieri,et al.  Interpretation of capillary zone electrophoresis compared with cellulose acetate and agarose gel electrophoresis: reference intervals and diagnostic efficiency in dogs and cats. , 2010, Veterinary clinical pathology.

[7]  Russell Scarpino,et al.  Reference intervals and relationships between health status, carapace length, body mass, and water temperature and concentrations of plasma total protein and protein electrophoretogram fractions in Atlantic loggerhead sea turtles and green turtles. , 2010, Journal of the American Veterinary Medical Association.

[8]  L. Brunnberg,et al.  Determination of plasma albumin concentration in healthy and diseased turtles: a comparison of protein electrophoresis and the bromcresol green dye-binding method. , 2010, Veterinary clinical pathology.

[9]  Y. Saco,et al.  Plasma protein electrophoresis of Trachemys scripta and Iguana iguana. , 2010, Veterinary clinical pathology.

[10]  R. Poppenga,et al.  COMPARISON OF BLOOD VALUES IN FORAGING, NESTING, AND STRANDED LOGGERHEAD TURTLES (CARETTA CARETTA) ALONG THE COAST OF GEORGIA, USA , 2009, Journal of wildlife diseases.

[11]  F. Crivellente,et al.  Analysis of mouse, rat, dog, marmoset, and human serum proteins by capillary electrophoresis: comparison with agarose gel electrophoresis. , 2008, Veterinary clinical pathology.

[12]  P. Eckersall Proteins, Proteomics, and the Dysproteinemias , 2008 .

[13]  C. Cray,et al.  Protein electrophoresis of psittacine plasma. , 2007, Veterinary clinical pathology.

[14]  R. Poppenga,et al.  BLOOD VALUES IN FREE-RANGING NESTING LEATHERBACK SEA TURTLES (DERMOCHELYS CORIACEA) ON THE COAST OF THE REPUBLIC OF GABON , 2006, Journal of zoo and wildlife medicine : official publication of the American Association of Zoo Veterinarians.

[15]  A. L. Jensen,et al.  Method comparison in the clinical laboratory. , 2006, Veterinary clinical pathology.

[16]  X. Bossuyt Advances in serum protein electrophoresis. , 2006, Advances in clinical chemistry.

[17]  J. Cristol,et al.  Evaluation of a capillary zone electrophoresis system versus a conventional agarose gel system for routine serum protein separation and monoclonal component typing. , 2006, Clinical laboratory.

[18]  John C. Gicking,et al.  Plasma Protein Electrophoresis of the Atlantic Loggerhead Sea Turtle, Carretta carretta , 2004 .

[19]  C. Cray,et al.  Protein Electrophoresis: A Tool for the Reptilian and Amphibian Practitioner , 2002 .

[20]  P. Lutz,et al.  ALTERED IN VITRO IMMUNE RESPONSES IN GREEN TURTLES (CHELONIA MYDAS) WITH FIBROPAPILLOMATOSIS , 2009, Journal of zoo and wildlife medicine : official publication of the American Association of Zoo Veterinarians.

[21]  G. Balazs,et al.  IMMUNE STATUS OF FREE-RANGING GREEN TURTLES WITH FIBROPAPILLOMATOSIS FROM HAWAII , 2001, Journal of wildlife diseases.