Characterisation of the normal canine serum proteome using a novel electrophoretic technique combined with mass spectrometry

Atherton, Matthew J.; Braceland, Mark; Harvie, Julie; Burchmore, Richard; Eadie, S.; Eckersall, P.D.; Morris, Joanna S.

doi:10.1016/j.tvjl.2012.12.011

Cited by 16 publications

(8 citation statements)

References 21 publications

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“…Electrophoretic migration patterns have been reported for 22 of these for several species (including the horse, another Perissodactyl) which are shown in Table 3 . Regarding the potential α1b band, seven of the identified proteins have been described to migrate to the α1 fraction, 11 to the α2 fraction and four to the β fraction; and in the potential α2 band, five proteins have been reported to migrate to α1, 13 to α2 and nine to β [ 12 – 15 , 28 – 30 ]. The majority of proteins identified in both bands were therefore α-globulins, and this was taken as confirmation that these bands represented α rather than β fractions.…”

Section: Resultsmentioning

confidence: 99%

“…Other methods can be used in order to more accurately identify the fractions. These are based on the identification of specific proteins within fractions and include exchange chromatography, special stains, and mass spectrometry [ 14 , 15 , 28 , 42 ]. For our study, the designation of the third, large globulin fraction as α2- rather than β1-globulin was confirmed by the finding, through proteomic analysis, that the majority of proteins within this fraction have been described to have an α-globulin migration in other species, including the horse.…”

Section: Discussionmentioning

confidence: 99%

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Serum protein electrophoresis in healthy and injured southern white rhinoceros (Ceratotherium simum simum)

et al. 2018

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Investigation of globulin fractions by serum protein electrophoresis (SPE) is the first step towards evaluation of the proteome in the southern white rhinoceros (Ceratotherium simum simum). Furthermore, identification of changes in globulins in animals with poaching and other injuries can guide discovery of potentially useful biomarkers of inflammation. The aim of this study was to develop reference intervals for agarose gel SPE in healthy white rhinoceros and to compare these serum protein electrophoresis results to those from animals with tissue trauma. Reference intervals for total serum protein and agarose gel electrophoretic albumin and globulin fractions were generated using serum samples from 49 healthy free-ranging adult white rhinoceros. A standardised gating system together with identification of specific proteins by mass spectrometry aided in fraction identification. Six globulin fractions were identified: α1a, α1b, α2, β1, β2 and γ. Reference intervals were generated for total serum protein (76–111 g/L), albumin (10–27 g/L) and globulin fractions (α1a: 1.6–3.2 g/L; α1b: 1.7–3.6 g/L; α2: 16.1–26.6 g/L; β1: 6.6–18.2 g/L; β2: 11.8–30.4 g/L; γ: 10.4–23.1 g/L; albumin: globulin ratio: 0.12–0.39). Results were compared to those from 30 animals with various degrees and chronicities of tissue trauma. Wounded animals had lower concentrations of total serum protein, albumin, total globulin, α and β1 globulins, lower percentages of α2 and β1 globulins, and higher percentages of β2 and γ globulins. These protein changes are similar to those seen in human patients with wounds rather than classic acute phase or chronic inflammatory responses.

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Section: Resultsmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Serum protein electrophoresis in healthy and injured southern white rhinoceros (Ceratotherium simum simum)

et al. 2018

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“…Agarose gel electrophoresis and CAE are available as manual or semi‐automatic methods. These methods use a porous solid matrix to help separate the protein fractions based on size and charge and stabilize the proteins for subsequent staining and evaluation . After electrophoresis, the proteins in the sample are stained with Amido black, Coomassie blue, or another stain which will label all proteins.…”

Section: Methodsmentioning

confidence: 99%

“…In most settings, chemically determined albumin is expected to be higher than densitometric albumin, due to binding of the bromocresol green dye to α‐ and β‐globulins . Additionally, some albumin has been shown to electrophoretically migrate in the globulin fraction of several species . M‐proteins can cause a significant falsely increased or decreased colorimetric albumin concentration, and in cases where the M‐protein is not soluble under the conditions of the total protein measurement, M‐proteins can cause falsely decreased total protein measurements (Figure ) …”

Section: Methodsmentioning

confidence: 99%

Protein characterization using electrophoresis and immunofixation; a case‐based review of dogs and cats

Moore

Avery

2019

Veterinary Clinical Pathol

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Protein electrophoresis and immunotyping can be a useful adjunct to the standard biochemical techniques for characterizing serum and urine proteins. This paper reviews currently available and commonly used methods for diagnostic protein electrophoresis, including both agarose gel and capillary zone electrophoretic techniques and total protein assessments. Immunofixation and immunosubtraction methods for identification of immunoglobulin location and class are also presented. Practical application of quality assurance and quality control strategies in compliance with American Society of Veterinary Clinical Pathology (ASVCP) best practices are discussed. Commonly encountered serum and urine electrophoretic diagnostic patterns, including electrophoretically normal, acute‐phase protein responses, polyclonal gammopathies, restricted polyclonal/oligoclonal gammopathies, paraproteinemias (monoclonal or biclonal gammopathies), and Bence‐Jones proteinurias are also reviewed using relevant case material. Cases in which immunofixation electrophoresis are particularly useful are highlighted, and methodologies to more accurately quantify serum monoclonal proteins (M‐proteins), monitoring tests commonly used in human medicine, are discussed.

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“…Although SPE has been described as the preferred reference method for the measurement of albumin and globulin, the true accuracy of this method is also questionable. Albumin was found to be present in the α1‐fraction of cats and dogs and the α1‐ and α2‐fractions of the white rhinoceros, indicating that albumin concentrations determined by calculation from the SPE albumin fraction are an underestimation of this protein . In addition, SPE albumin determination could be inaccurate due to disproportional staining of the albumin band because of the high affinity of albumin for the dye, or non‐linearity of light absorbance during densitometric measurements .…”

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confidence: 99%

Bias between two methods of albumin measurement in the white rhinoceros, Ceratotherium simum

Hooijberg

Cray

Miller

et al. 2020

Veterinary Clinical Pathol

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Background The bromocresol green (BCG) method has been reported to overestimate serum albumin concentration in several species due to non‐specific binding to globulins. As the white rhinoceros has high concentrations of serum globulins, significant differences in albumin measured by the BCG method, and the field method of agarose gel serum protein electrophoresis (SPE) are expected. Objectives We aimed to compare the BCG and SPE methods for albumin determination in the serum of white rhinoceroses. Methods SPE and BCG albumin were measured in 82 white rhinoceros serum samples. Results were compared using Bland‐Altman difference plots and Passing‐Bablok regression analysis. Results BCG albumin showed a significant mean constant positive bias of 7 g/L, or 36%, which was more than the total allowable error of 15% and was clinically significant. Methods were not comparable within the inherent imprecision of each method. Conclusions The BCG method overestimated albumin concentrations in this species compared with agarose gel SPE, and method‐specific reference intervals should be used.

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Characterisation of the normal canine serum proteome using a novel electrophoretic technique combined with mass spectrometry

Cited by 16 publications

References 21 publications

Serum protein electrophoresis in healthy and injured southern white rhinoceros (Ceratotherium simum simum)

Serum protein electrophoresis in healthy and injured southern white rhinoceros (Ceratotherium simum simum)

Protein characterization using electrophoresis and immunofixation; a case‐based review of dogs and cats

Bias between two methods of albumin measurement in the white rhinoceros, Ceratotherium simum

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