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患有和未患淋巴瘤的猫的琼脂糖凝胶血清蛋白电泳及串联质谱指纹图谱分析的初步结果

Agarose gel serum protein electrophoresis in cats with and without lymphoma and preliminary results of tandem mass fingerprinting analysis.

作者信息

Gerou-Ferriani Magda, McBrearty Alix R, Burchmore Richard J, Jayawardena Kamburapola G I, Eckersall P David, Morris Joanna S

机构信息

School of Veterinary Medicine, University of Glasgow, Glasgow, UK.

出版信息

Vet Clin Pathol. 2011 Jun;40(2):159-73. doi: 10.1111/j.1939-165X.2011.00310.x. Epub 2011 May 10.

Abstract

BACKGROUND

Serum electrophoretic profiles in cats are poorly characterized with respect to the proteins that comprise the globulin fractions, and interpretation of the electrophoretograms is routinely done in the absence of information about identity of the proteins found within each fraction.

OBJECTIVES

The aims of this study were to compare protein fractions separated by serum protein electrophoresis (SPE) in healthy cats and in cats with lymphoma and to confirm some component proteins in the major fractions following SPE using tandem mass fingerprinting analysis (TMFA).

METHODS

Total protein concentration was measured and agarose gel SPE performed on serum from 14 healthy cats and 14 cats with lymphoma. The absolute protein concentration within each fraction was compared between the 2 groups. Bands corresponding to the SPE fractions were excised from the gels of 2 control cats and 1 cat with lymphoma and analyzed by liquid chromatography coupled to mass spectrometry. Results were compared with sequences in the National Center for Biotechnology Information protein database.

RESULTS

Median albumin concentrations were significantly decreased and median β-globulin concentrations were significantly increased in cats with lymphoma. Narrow electrophoretic spikes were present in the β/γ-globulin fraction in 3 cats with lymphoma. Following TMFA, multiple proteins were identified in each fraction, and their mobility agreed with results from previous studies generated using alternative techniques. Inter-α (globulin) inhibitor 4 was identified in feline serum for the first time.

CONCLUSIONS

Cats with lymphoma had lower albumin and higher β-globulin concentrations than did healthy cats. Despite limitations of one-dimensional agarose gel SPE, TMFA provided preliminary data to confirm the protein components of the various fractions.

摘要

背景

关于构成猫球蛋白组分的蛋白质,猫血清电泳图谱的特征描述不足,并且在缺乏各组分中蛋白质身份信息的情况下,通常对电泳图进行解读。

目的

本研究旨在比较健康猫和淋巴瘤猫血清蛋白电泳(SPE)分离出的蛋白质组分,并使用串联质谱指纹分析(TMFA)确认SPE后主要组分中的一些组成蛋白。

方法

测量14只健康猫和14只淋巴瘤猫血清的总蛋白浓度并进行琼脂糖凝胶SPE。比较两组中各组分内的绝对蛋白浓度。从2只对照猫和1只淋巴瘤猫的凝胶中切下与SPE组分相对应的条带,通过液相色谱-质谱联用进行分析。将结果与美国国立生物技术信息中心蛋白质数据库中的序列进行比较。

结果

淋巴瘤猫的白蛋白中位数浓度显著降低,β球蛋白中位数浓度显著升高。3只淋巴瘤猫的β/γ球蛋白组分中出现窄电泳峰。经过TMFA分析,在每个组分中鉴定出多种蛋白质,其迁移率与先前使用其他技术得出的研究结果一致。首次在猫血清中鉴定出α(球蛋白)间抑制因子4。

结论

与健康猫相比,淋巴瘤猫的白蛋白浓度较低,β球蛋白浓度较高。尽管一维琼脂糖凝胶SPE存在局限性,但TMFA提供了初步数据以确认各组分的蛋白质成分。

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