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Development of a Solid-Phase Colorimetric Assay for the Screening of Transglutaminase Activities

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Authors
Choi, Jeeyoung; Choi, Gyung Ho; Woo, Kang Mee; Park, Sang Chul
Issue Date
1992-06
Publisher
Seoul National University College of Medicine
Citation
Seoul J Med 1992;33(2):167-173
Keywords
TransglutaminasesAssayScreeningDevelopment
Abstract
A solid-phase colorimetric assay method for transglutaminase activities
has been developed. The principle of the assay is to monitor cross-linking activities between
casein bound to microtiter plates and free biotinylated casein by the sample.
Quantitation of immobilized biotin-labeled casein formation through the enzymatic reaction
was conducted by avidin or streptavidin conjugated enzymes. For this purpose,
the efficiency of four different reporter enzymes (streptavidin or avidin conjugated alkaline
phosphatase, streptavidin or avidin conjugated horseradish peroxidase) was
compared, especially focusing on the sensitivity and specificity of the respective
methods. The newly developed assay method was applied to the procedure of
transglutaminase C purification from human erythrocytes and proven to have good correlation
with conventional C14 putrescine method (r = 0.85, p(0.05). Moreover, since
this new method can detect enzymatic activity without use of radioisotope and can process
a number of samples simultaneously, it is possible to screen a mass population for
transglutaminase deficiency, such as factor XIII, in routine clinical laboratories.
ISSN
0582-6802
Language
English
URI
http://hdl.handle.net/10371/6280
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Appears in Collections:
College of Medicine/School of Medicine (의과대학/대학원)Dept. of Medicine (의학과)The Seoul Journal of MedicineThe Seoul Journal of Medicine Vol. 33 No.3 (1992)
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