ELISA-based method to detect microbial transglutaminase in frozen surimi

被引：0

作者：

Li, Yi ^{[1
]}

Fan, Da-Ming ^{[1
]}

Gu, Zhen-Nan ^{[1
]}

Huang, Jian-Lian ^{[2
]}

Lian, Hui-Zhang ^{[3
]}

Zhao, Jian-Xin ^{[1
]}

Chen, Wei ^{[1
]}

Zhang, Hao ^{[1
]}

机构：

[1] Staten Key Laboratory of Food Science and Technology, School of Food Science and Technology, Jiangnan University, Wuxi

[2] Fujian Anjoyfood Share Co. Ltd., Xiamen

[3] Wuxi Huashunminsheng Food Co. Ltd., Wuxi

来源：

Modern Food Science and Technology | 2014年 / 30卷 / 09期

关键词：

Frozen surimi; Microbial transglutaminase; Sandwich-enzyme-linked immunosorbent assay detection;

D O I：

10.13982/j.mfst.1673-9078.2014.09.045

中图分类号：

学科分类号：

摘要：

In this study, a sandwich-type enzyme-linked immunosorbent assay (ELISA) was developed to detect microbial transglutaminase (MTG) in frozen surimi, using rabbit anti-MTG polyclonal antibody as the capture antibody and mouse anti-MTG monoclonal antibody as the detection antibody. Combined with anti-IgG1 secondary antibody horseradish peroxidase (HRP) conjugate, the sandwich ELISA was developed to quantitatively detect MTG in frozen surimi samples, thus providing a means to prevent adulteration of frozen surimi. The results showed that the optimum working concentration of the capture antibody was 2 µg/mL whereas that of the detection antibody was 0.1 µg/mL, and the optimum dilution of HRP-conjugated secondary antibody was 1:5,000. The visual detection limit of the assay was 20 ng/mL, and alinear relationship between OD450 values and the log values of MTG concentration was observed in the range of 0.6~10 mg/mL. A simulation model of adulteration in frozen surimi was established to determine the amount of added MTG. The recovery rate of MTG was over 94%, whereas the intra- and inter-plate coefficients of variation during the measurement were 1.12%~4.02% and 5.43%~6.87%, respectively. These results proved that this method was sensitive, required simple sample pretreatment, and was suitable for the quantitative detection of MTG added to frozen surimi.

引用

页码：275 / 279

页数：4

共 13 条

[1]

Kuraishi C., Yamazaki K., Susa Y., Transglutaminase: its utilization in the food industry , Food Reviews International, 17, pp. 221-246, (2001)

[2]

Yokoyama K., Nio N., Kikuchi Y., Properties and applications of microbial transglutaminase , Applied Microbiology and Biotechnology, 64, pp. 447-454, (2004)

[3]

Bao Y.-L., Pan L., Research progress of mechanism and activity assay of microbial transglutaminase , Science and Technology of Food Industry, 29, 7, pp. 265-269, (2008)

[4]

Zhang W.-X., Immunology Experimental Techniques , (2007)

[5]

Wang Y.-H., Li G.-C., Zhou X.-F., Antibodies Theory and Technology , (2004)

[6]

Kaufmann A., Koppel R., Widmer M., Determination of microbial transglutaminase in meat and meat products , Food Additives & Contaminants: Part A, 29, 9, pp. 1364-1373, (2012)

[7]

Ohtsuka T., Seguro K., Motoki M., Microbial transglutaminase estimation in enzyme-treated surimi-based products by enzyme immunosorbentassay , Journal of Food Science, 61, 1, pp. 81-84, (1996)

[8]

Schloegl W., Klein A., Et al., Residual transglutaminase in collagen-effects, detection, quantification, and removal , European Journal of Pharmaceutics and Biopharmaceutics, 80, 2, pp. 282-288, (2012)

[9]

Engvall E., Perlmann P., Enzyme-linked immunosorbent assay (ELISA) quantitative assay of immunoglobulin G , Immunochemistry, 8, 9, pp. 871-874, (1971)

[10]

Loizou S., McCrea J.D., Rudge A.C., Et al., Measurement of anti-cardiolipin antibodies by an enzyme-linked immunosorbent assay (ELISA): standardization and quantitation of results , Clinical and Experimental Immunology, 62, 3, pp. 738-745, (1985)

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